Novel proteins and nucleic acids encoding same

BACKGROUND OF THE INVENTION

[0002] The invention generally relates to nucleic acids and polypeptides. More particularly, the invention relates to nucleic acids encoding novel G-protein coupled receptor (GPCR) polypeptides, as well as vectors, host cells, antibodies, and recombinant methods for producing these nucleic acids and polypeptides.

SUMMARY OF THE INVENTION

[0003] The invention is based in part upon the discovery of nucleic acid sequences encoding novel polypeptides. The novel nucleic acids and polypeptides are referred to herein as “GPCRX” nucleic acids and polypeptides. These nucleic acids and polypeptides, as well as derivatives, homologs, analogs and fragments thereof, will hereinafter be collectively designated as “GPCRX” nucleic acid or polypeptide sequences.

[0004] In one aspect, the invention provides an isolated GPCRX nucleic acid molecule encoding a GPCRX polypeptide that includes a nucleic acid sequence that has identity to the nucleic acids disclosed in SEQ ID NOS:1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, 75, 77, 79, 81, 83, 85, 87, 89, 91, 93, 95, 97, 99, 101, 103, 105, 107, 109, 111, 113, 115, 117, 119, 121, 123, 125, 127, 129, 131, 133, 135, 137, 139, 141, 143, 145, 147, 149 and 151. In some embodiments, the GPCRX nucleic acid molecule will hybridize under stringent conditions to a nucleic acid sequence complementary to a nucleic acid molecule that includes a protein-coding sequence of a GPCRX nucleic acid sequence. The invention also includes an isolated nucleic acid that encodes a GPCRX polypeptide, or a fragment, homolog, analog or derivative thereof. For example, the nucleic acid can encode a polypeptide at least 80% identical to a polypeptide comprising the amino acid sequences of SEQ ID NOS: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, 78, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 100, 102, 104, 106, 108, 110, 112, 114, 116, 118, 120, 122, 124, 126, 128, 130, 132, 134, 136, 138, 140, 142, 144, 146, 148, 150 and 152. The nucleic acid can be, for example, a genomic DNA fragment or a cDNA molecule that includes the nucleic acid sequence of any of SEQ ID NOS:1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, 75, 77, 79, 81, 83, 85, 87, 89, 91, 93, 95, 97, 99, 101, 103, 105, 107, 109, 111, 113, 115, 117, 119, 121, 123, 125, 127, 129, 131, 133, 135, 137, 139, 141, 143, 145, 147, 149 and 151.

[0005] Also included in the invention is an oligonucleotide, e.g., an oligonucleotide which includes at least 6 contiguous nucleotides of a GPCRX nucleic acid (e.g., SEQ ID NOS:1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, 75, 77, 79, 81, 83, 85, 87, 89, 91, 93, 95, 97, 99, 101, 103, 105, 107, 109, 111, 113, 115, 117, 119, 121, 123, 125, 127, 129, 131, 133, 135, 137, 139, 141, 143, 145, 147, 149 and 151) or a complement of said oligonucleotide.

[0006] Also included in the invention are substantially purified GPCRX polypeptides (e.g., SEQ ID NOS: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, 78, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 100, 102, 104, 106, 108, 110, 112, 114, 116, 118, 120, 122, 124, 126, 128, 130, 132, 134, 136, 138, 140, 142, 144, 146, 148, 150 and 152). In certain embodiments, the GPCRX polypeptides include an amino acid sequence that is substantially identical to the amino acid sequence of a human GPCRX polypeptide.

[0007] The invention also features antibodies that immunoselectively bind to GPCRX polypeptides, or fragments, homologs, analogs or derivatives thereof.

[0008] In another aspect, the invention includes pharmaceutical compositions that include therapeutically- or prophylactically-effective amounts of a therapeutic and a pharmaceutically-acceptable carrier. The therapeutic can be, e.g., a GPCRX nucleic acid, a GPCRX polypeptide, or an antibody specific for a GPCRX polypeptide. In a further aspect, the invention includes, in one or more containers, a therapeutically- or prophylactically-effective amount of this pharmaceutical composition.

[0009] In a further aspect, the invention includes a method of producing a polypeptide by culturing a cell that includes a GPCRX nucleic acid, under conditions allowing for expression of the GPCRX polypeptide encoded by the DNA. If desired, the GPCRX polypeptide can then be recovered.

[0010] In another aspect, the invention includes a method of detecting the presence of a GPCRX polypeptide in a sample. In the method, a sample is contacted with a compound that selectively binds to the polypeptide under conditions allowing for formation of a complex between the polypeptide and the compound. The complex is detected, if present, thereby identifying the GPCRX polypeptide within the sample.

[0011] The invention also includes methods to identify specific cell or tissue types based on their expression of a GPCRX.

[0012] Also included in the invention is a method of detecting the presence of a GPCRX nucleic acid molecule in a sample by contacting the sample with a GPCRX nucleic acid probe or primer, and detecting whether the nucleic acid probe or primer bound to a GPCRX nucleic acid molecule in the sample.

[0013] In a further aspect, the invention provides a method for modulating the activity of a GPCRX polypeptide by contacting a cell sample that includes the GPCRX polypeptide with a compound that binds to the GPCRX polypeptide in an amount sufficient to modulate the activity of said polypeptide. The compound can be, e.g., a small molecule, such as a nucleic acid, peptide, polypeptide, peptidomimetic, carbohydrate, lipid or other organic (carbon containing) or inorganic molecule, as further described herein.

[0014] Also within the scope of the invention is the use of a therapeutic in the manufacture of a medicament for treating or preventing disorders or syndromes including, e.g., developmental diseases; MHCII and III diseases (immune diseases); taste and scent detectability disorders; Burkitt's lymphoma; corticoneurogenic disease; signal transduction pathway disorders; metabolic pathway disorders; retinal diseases including those involving photoreception; cell growth rate disorders; cell shape disorders; metabolic disorders; feeding disorders; control of feeding; the metabolic syndrome X; wasting disorders associated with chronic diseases; obesity; potential obesity due to over-eating or metabolic disturbances; potential disorders due to starvation (lack of appetite); diabetes; noninsulin-dependent diabetes mellitus (NIDDM1); infectious disease; bacterial, fungal, protozoal and viral infections (particularly infections caused by HIV-1 or HIV-2); pain; cancer (including but not limited to neoplasm; adenocarcinoma; lymphoma; prostate cancer; uterus cancer); cancer-associated cachexia; anorexia; bulimia; asthma; Parkinson's disease; acute heart failure; hypotension; hypertension; urinary retention; osteoporosis; Crohn's disease; multiple sclerosis; Albright Hereditary Ostoeodystrophy; angina pectoris; myocardial infarction; ulcers; allergies; benign prostatic hypertrophy; and psychotic and neurological disorders; including anxiety; schizophrenia; manic depression; delirium; dementia; neurodegenerative disorders; Alzheimer's disease; severe mental retardation; Dentatorubro-pallidoluysian atrophy (DRPLA); Hypophosphatemic rickets; autosomal dominant (2) Acrocallosal syndrome and dyskinesias, such as Huntington's disease or Gilles de la Tourette syndrome; immune disorders; Adrenoleukodystrophy; Congenital Adrenal Hyperplasia; Hemophilia; Hypercoagulation; Idiopathic thrombocytopenic purpura; autoimmume disease; immunodeficiencies; transplantation; Von Hippel-Lindau (VHL) syndrome; Stroke; Tuberous sclerosis; hypercalceimia; Cerebral palsy; Epilepsy; Lesch-Nyhan syndrome; Ataxia-telangiectasia; Leukodystrophies; Behavioral disorders; Addiction; Neuroprotection; Cirrhosis; Transplantation; Systemic lupus erythematosus; Emphysema; Scleroderma; ARDS; Renal artery stenosis; Interstitial nephritis; Glomerulonephritis; Polycystic kidney disease; Systemic lupus erythematosus; Renal tubular acidosis; IgA nephropathy; Cardiomyopathy; Atherosclerosis; Congenital heart defects; Aortic stenosis; Atrial septal defect (ASD); Atrioventricular (A-V) canal defect; Ductus arteriosus; Pulmonary stenosis; Subaortic stenosis; Ventricular septal defect (VSD); valve diseases; Scleroderma; fertility; Pancreatitis; Endocrine dysfunctions; Growth and reproductive disorders; Inflammatory bowel disease; Diverticular disease; Leukodystrophies; Graft vesus host; Hyperthyroidism; Endometriosis; hematopoietic disorders and/or other pathologies and disorders of the like. The therapeutic can be, e.g., a GPCRX nucleic acid, a GPCRX polypeptide, or a GPCRX-specific antibody, or biologically-active derivatives or fragments thereof.

[0015] For example, the compositions of the present invention will have efficacy for treatment of patients suffering from the diseases and disorders listed above and/or other pathologies and disorders.

[0016] The polypeptides can be used as immunogens to produce antibodies specific for the invention, and as vaccines. They can also be used to screen for potential agonist and antagonist compounds. For example, a cDNA encoding GPCRX may be useful in gene therapy, and GPCRX may be useful when administered to a subject in need thereof. By way of nonlimiting example, the compositions of the present invention will have efficacy for treatment of patients suffering the diseases and disorders listed above and/or other pathologies and disorders.

[0017] The invention further includes a method for screening for a modulator of disorders or syndromes including, e.g., diseases and disorders listed above and/or other pathologies and disorders and those disorders related to cell signal processing and metabolic pathway modulation. The method includes contacting a test compound with a GPCRX polypeptide and determining if the test compound binds to said GPCRX polypeptide. Binding of the test compound to the GPCRX polypeptide indicates the test compound is a modulator of activity, or of latency or predisposition to the aforementioned disorders or syndromes.

[0018] Also within the scope of the invention is a method for screening for a modulator of activity, or of latency or predisposition to an disorders or syndromes including the diseases and disorders listed above and/or other pathologies and disorders or other disorders related to cell signal processing and metabolic pathway modulation by administering a test compound to a test animal at increased risk for the aforementioned disorders or syndromes. The test animal expresses a recombinant polypeptide encoded by a GPCRX nucleic acid. Expression or activity of GPCRX polypeptide is then measured in the test animal, as is expression or activity of the protein in a control animal which recombinantly-expresses GPCRX polypeptide and is not at increased risk for the disorder or syndrome. Next, the expression of GPCRX polypeptide in both the test animal and the control animal is compared. A change in the activity of GPCRX polypeptide in the test animal relative to the control animal indicates the test compound is a modulator of latency of the disorder or syndrome.

[0019] In yet another aspect, the invention includes a method for determining the presence of or predisposition to a disease associated with altered levels of a GPCRX polypeptide, a GPCRX nucleic acid, or both, in a subject (e.g., a human subject). The method includes measuring the amount of the GPCRX polypeptide in a test sample from the subject and comparing the amount of the polypeptide in the test sample to the amount of the GPCRX polypeptide present in a control sample. An alteration in the level of the GPCRX polypeptide in the test sample as compared to the control sample indicates the presence of or predisposition to a disease in the subject. Preferably, the predisposition includes diseases and disorders listed above and/or other pathologies and disorders. Also, the expression levels of the new polypeptides of the invention can be used in a method to screen for various cancers as well as to determine the stage of cancers.

[0020] In a further aspect, the invention includes a method of treating or preventing a pathological condition associated with a disorder in a mammal by administering to the subject a GPCRX polypeptide, a GPCRX nucleic acid, or a GPCRX-specific antibody to a subject (e.g., a human subject), in an amount sufficient to alleviate or prevent the pathological condition. In preferred embodiments, the disorder, includes the diseases and disorders listed above and/or other pathologies and disorders.

[0021] In yet another aspect, the invention can be used in a method to identity the cellular receptors and downstream effectors of the invention by any one of a number of techniques commonly employed in the art. These include but are not limited to the two-hybrid system, affinity purification, co-precipitation with antibodies or other specific-interacting molecules.

[0022] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention, suitable methods and materials are described below. All publications, patent applications, patents, and other references mentioned herein are incorporated by reference in their entirety. In the case of conflict, the present specification, including definitions, will control. In addition, the materials, methods, and examples are illustrative only and not intended to be limiting.

[0023] Other features and advantages of the invention will be apparent from the following detailed description and claims.

DETAILED DESCRIPTION OF THE INVENTION

[0024] The invention is based, in part, upon the discovery of novel nucleic acid sequences that encode novel polypeptides. The novel nucleic acids and their encoded polypeptides are collectively designated herein as “GPCRX”.

[0025] The novel GPCRX nucleic acids of the invention include the nucleic acids whose sequences are provided in Table 1, inclusive, or a fragment, derivative, analog or homolog thereof. The novel GPCRX proteins of the invention include the protein fragments whose sequences are provided in Table 1, inclusive. The individual GPCRX nucleic acids and proteins are described below. Within the scope of this invention is a method of using these nucleic acids and peptides in the treatment or prevention of a disorder related to cell signaling or metabolic pathway modulation.

[0026] The GPCRX proteins of the invention have a high homology to the 7tm—1 domain (PFam Acc. No. pfam00001). The 7tm—1 domain is from the 7 transmembrane receptor family, which includes a number of different proteins, including, for example, serotonin receptors, dopamine receptors, histamine receptors, andrenergic receptors, cannabinoid receptors, angiotensin II receptors, chemokine receptors, opioid receptors, G-protein coupled receptor (GPCR) proteins, olfactory receptors (OR), and the like. Some proteins and the Protein Data Base Ids/gene indexes include, for example: rhodopsin (129209); 5-hydroxytryptamine receptors; (112821, 8488960, 112805, 231454, 1168221, 398971, 112806); G protein-coupled receptors (119130, 543823, 1730143, 132206, 137159, 6136153, 416926, 1169881, 136882, 134079); gustatory receptors (544463, 462208); c-x-c chemokine receptors (416718, 128999, 416802, 548703, 1352335); opsins (129193, 129197, 129203); and olfactory receptor-like proteins (129091, 1171893, 400672, 548417).

[0027] Because of the close homology among the members of the GPCRX family, proteins that are homologous to any one member of the family are also largely homologous to the other members, except where the sequences are different as shown below.

[0028] The similarity information for the GPCRX proteins and nucleic acids disclosed herein suggest that GPCRX may have important structural and/or physiological functions characteristic of the Olfactory Receptor family and the GPCR family. Therefore, the nucleic acids and proteins of the invention are useful in potential diagnostic and therapeutic applications and as a research tool. These include serving as a specific or selective nucleic acid or protein diagnostic and/or prognostic marker, wherein the presence or amount of the nucleic acid or the protein are to be assessed, as well as potential therapeutic applications such as the following: (i) a protein therapeutic, (ii) a small molecule drug target, (iii) an antibody target (therapeutic, diagnostic, drug targeting/cytotoxic antibody), (iv) a nucleic acid useful in gene therapy (gene delivery/gene ablation), and (v) a composition promoting tissue regeneration in vitro and in vivo (vi) biological defense weapon.

[0029] G-Protein Coupled Receptor proteins (“GPCRs”) have been identified as a large family of G protein-coupled receptors in a number of species. These receptors share a seven transmembrane domain structure with many neurotransmitter and hormone receptors, and are likely to underlie the recognition and G-protein-mediated transduction of various signals. Human GPCR generally do not contain introns and belong to four different gene subfamilies, displaying great sequence variability. These genes are dominantly expressed in olfactory epithelium. See, e.g., Ben-Arie et al., Hum. Mol Genet. 1994 3:229-235; and, Online Mendelian Inheritance in Man (“OMIM”) entry # 164342 (http://www.ncbi.nlm.nih.gov/entrez/dispomim.cgi?).

[0030] The olfactory receptor (“OR”) gene family constitutes one of the largest GPCR multigene families and is distributed among many chromosomal sites in the human genome. See Rouquier et al., Hum. Mol. Genet. 7(9): 1337-45 (1998); Malnic et al., Cell 96:713-23 (1999). Olfactory receptors constitute the largest family among G protein-coupled receptors, with up to 1000 members expected. See Vanderhaeghen et al., Genomics 39(3):23946 (1997); Xie et al., Mamm. Genome 11 (12):1070-78 (2000); Issel-Tarver et al., Proc. Natl. Acad. Sci. USA 93(20): 10897-902 (1996). The recognition of odorants by olfactory receptors is the first stage in odor discrimination. See Krautwurst et al., Cell 95(7):917-26 (1998); Buck et al., Cell 65(1):175-87 (1991). Many ORs share some characteristic sequence motifs and have a central variable region corresponding to a putative ligand binding site. See Issel-Tarver et al., Proc. Natl. Acad. Sci. USA 93:10897-902 (1996).

[0031] Other examples of seven membrane spanning proteins that are related to GPCRs are chemoreceptors. See Thomas et al., Gene 178(1-2):1-5 (1996). Chemoreceptors have been identified in taste, olfactory, and male reproductive tissues. See id.; Walensky et al., J. Biol. Chem. 273(16):9378-87 (1998); Parmentier et al., Nature 355(6359):453-55 (1992); Asai et al., Biochem. Biophys. Res. Commun. 221(2):240-47 (1996).

[0032] The GPCRX nucleic acids of the invention encoding GPCR-like proteins include the nucleic acids whose sequences are provided herein, or fragments thereof. The invention also includes mutant or variant nucleic acids any of whose bases may be changed from the corresponding base shown herein while still encoding a protein that maintains its GPCR-like activities and physiological functions, or a fragment of such a nucleic acid. The invention further includes nucleic acids whose sequences are complementary to those just described, including nucleic acid fragments that are complementary to any of the nucleic acids just described. The invention additionally includes nucleic acids or nucleic acid fragments, or complements thereto, whose structures include chemical modifications. Such modifications include, by way of nonlimiting example, modified bases, and nucleic acids whose sugar phosphate backbones are modified or derivatized. These modifications are carried out at least in part to enhance the chemical stability of the modified nucleic acid, such that they may be used, for example, as antisense binding nucleic acids in therapeutic applications in a subject.

[0033] The GPCRX proteins of the invention include the GPCR-like proteins whose sequences are provided herein. The invention also includes mutant or variant proteins any of whose residues may be changed from the corresponding residue shown herein while still encoding a protein that maintains its GPCR-like activities and physiological functions, or a functional fragment thereof. The invention further encompasses antibodies and antibody fragments, such as Fab or (Fab)2, that bind immunospecifically to any of the proteins of the invention.

[0034] The GPCRX nucleic acids and proteins are useful in potential therapeutic applications implicated in various GPCR-related pathological disorders and/or OR-related pathological disorders, described further below. For example, a cDNA encoding the GPCR (or olfactory-receptor) like protein may be useful in gene therapy, and the receptor-like protein may be useful when administered to a subject in need thereof. The nucleic acids and proteins of the invention are also useful in potential therapeutic applications used in the treatment of developmental diseases; MHCII and III diseases (immune diseases); taste and scent detectability disorders; Burkitt's lymphoma; corticoneurogenic disease; signal transduction pathway disorders; metabolic pathway disorders; retinal diseases including those involving pbotoreception; cell growth rate disorders; cell shape disorders; metabolic disorders; feeding disorders; control of feeding; the metabolic syndrome X; wasting disorders associated with chronic diseases; obesity; potential obesity due to over-eating or metabolic disturbances; potential disorders due to starvation (lack of appetite); diabetes; noninsulin-dependent diabetes mellitus (NIDDM1); infectious disease; bacterial, fungal, protozoal and viral infections (particularly infections caused by HIV-1 or HIV-2); pain; cancer (including but not limited to neoplasm; adenocarcinoma; lymphoma; prostate cancer; uterus cancer); cancer-associated cachexia; anorexia; bulimia; asthma; Parkinson's disease; acute heart failure; hypotension; hypertension; urinary retention; osteoporosis; Crohn's disease; multiple sclerosis; Albright Hereditary Ostoeodystrophy; angina pectoris; myocardial infarction; ulcers; allergies; benign prostatic hypertrophy; and psychotic and neurological disorders; including anxiety; schizophrenia; manic depression; delirium; dementia; neurodegenerative disorders; Alzheimer's disease; severe mental retardation; Dentatorubro-pallidoluysian atrophy (DRPLA); Hypophosphatemic rickets; autosomal dominant (2) Acrocallosal syndrome and dyskinesias, such as Huntington's disease or Gilles de la Tourette syndrome; immune disorders; Adrenoleukodystrophy; Congenital Adrenal Hyperplasia; Hemophilia; Hypercoagulation; Idiopathic thrombocytopenic purpura; autoimmume disease; immunodeficiencies; transplantation; Von Hippel-Lindau (VHL) syndrome; Stroke; Tuberous sclerosis; hypercalceimia; Cerebral palsy; Epilepsy; Lesch-Nyhan syndrome; Ataxia-telangiectasia; Leukodystrophies; Behavioral disorders; Addiction; Neuroprotection; Cirrhosis; Transplantation; Systemic lupus erythematosus; Emphysema; Scleroderma; ARDS; Renal artery stenosis; Interstitial nephritis; Glomerulonephritis; Polycystic kidney disease; Systemic lupus erythematosus; Renal tubular acidosis; IgA nephropathy; Cardiomyopathy; Atherosclerosis; Congenital heart defects; Aortic stenosis; Atrial septal defect (ASD); Atrioventricular (A-V) canal defect; Ductus arteriosus; Pulmonary stenosis; Subaortic stenosis; Ventricular septal defect (VSD); valve diseases; Scleroderma; fertility; Pancreatitis; Endocrine dysfunctions; Growth and reproductive disorders; Inflammatory bowel disease; Diverticular disease; Leukodystrophies; Graft vesus host; Hyperthyroidism; Endometriosis; hematopoietic disorders and/or other pathologies and disorders. Other GPCR-related diseases and disorders are contemplated.

[0035] The polypeptides can be used as immunogens to produce antibodies specific for the invention, and as vaccines. They can also be used to screen for potential agonist and antagonist compounds. For example, a cDNA encoding the GPCR-like protein may be useful in gene therapy, and the GPCR-like protein may be useful when administered to a subject in need thereof. By way of nonlimiting example, the compositions of the present invention will have efficacy for treatment of patients suffering the diseases and disorders listed above and/or other pathologies and disorders. The novel nucleic acid encoding GPCR-like protein, and the GPCR-like protein of the invention, or fragments thereof, may further be useful in diagnostic applications, wherein the presence or amount of the nucleic acid or the protein are to be assessed. These materials are further useful in the generation of antibodies that bind immunospecifically to the novel substances of the invention for use in therapeutic or diagnostic methods.

[0036] All of the sequence listed in Table 1 have a high degree of homology to known GPCR sequences. Exemplary homology for the sequences is provided in the provisional applications from which the present application claims priority. This homology data are incorporated herein by reference in their entirety.

1TABLE 1DNA SEQID NO/PROTEINAcc. No.SEQ ID NODNA SEQUENCEPROTEIN SEQUENCEGMAP000916_A1/2AATGGCTGCAGGAAATCACTCTACAGTGACAGAGTTCATTCTCAAGGGTTTAACGAAMAAGNHSTVTEFILKGLTKRAGAGAGCAGACCTCCAGCTCCCCCTCTTTCTCCTCTTCCTCGGGATCTACTTGGTCACDLQLPLFLLFLGIYLVTIVGNCATCGTGGGGAACCTGGGCATGATCACTCTAATTTGTCTGAACTCTCAGCTGCACACLGMITLICLNSQLHTPMYYFLCCCCATGTACTACTTTCTCAGCAATCTGTCACTCATGGATCTCTGCTACTCCTCCGTSNLSLMDLCYSSVITPKMLVNCATTACCCCTAAGATGCTGGTGAACTTTGTGTCAGAGAAAAACATCATCTCCTACGCFVSEKNIISYAGCMSQLYFFLAGGGTGCATGTCACAGCTCTACTTCTTCCTTGTTTTTGTCATTGCTGAGTGTTACATVFVIAECYMLTVMAYDRYVAIGCTGACAGTGATGGCCTACGACCGCTATGTTGCCATCTGCCACCCTTTGCTTTACAACHPLLYNIIMSHHTCLLLVAVCATCATTATGTCTCATCACACCTGCCTGCTGCTGGTGGCTGTGGTCTACGCCATCGGVYAIGLIGSTIETGLMLKLPYACTCATTGGCTCCACAATAGAAACTGGCCTCATGTTAAAACTGCCCTATTGTGAGCACEHLISHYFCDILPLMKLSCSCCTCATCAGTCACTACTTCTGTGACATCCTCCCTCTCATGAAGCTGTCCTGCTCTAGSTYDVEMTVFFSAGFNIIVTSCACCTATGATGTTGAGATGACAGTCTTCTTTTCGGCTGGATTCAACATCATAGTCACLTVLVSTYFILSSILGISTTEGAGCTTAACAGTTCTTGTTTCTTACACCTTCATTCTCTCCAGCATCCTCGGCATCAGGRSKAFSTCSSHLAAVGMFYGCACCACAGAGGGGAGATCCAAAGCCTTCAGCACCTGCAGCTCCCACCTTGCAGCCGTSTAFMYLKPSTISSLTQENVAGGGAATGTTCTATGGATCAACTGCATTCATGTACTTAAAACCCTCCACAATCAGTTCSVFYTTVIPMLNPLIYSLRNKCTTGACCCAGGAGAATGTGGCCTCTGTGTTCTACACCACGGTAATCCCCATGTTGAAEVKAAVQKTLRGKLFTCCCCTAATCTACAGCCTGAGGAACAAGGAAGTAAAGGCTGCCGTGCAGAAAACGCTGAGGGGTAAACTGTTTTGATGCAAATGMAP001524_A3/4AATGGCTGCAGGAAATCACTCTACAGTGACAGAGTTCATTCTCAAGGGTTTAACGAAMAAGNHSTVTEFILKGLTKRAGAGAGCAGACCTCCAGCTCCCCCTCTTTCTCCTCTTCCTCGGGATCTACTTGGTCACDLQLPLFLLFLGIYLVTIVGNCATCGTGGGGAACCTGGGCATGATCACTCTAATTTGTCTGAACTCTCAGCTGCACACLGMITLICLNSQLHTPMYYFLCCCCATGTACTACTTTCTCAGCAATCTGTCACTCATGGATCTCTGCTACTCCTCCGTSNLSLMDLCYSSVITPLMLVNCATTACCCCTAAGATGCTGGTGAACTTTGTGTCAGAGAAAAACATCATCTCCTACGCFVSEKNIISYAGCMSQLYFFLAGGGTGCATGTCACAGCTCTACTTCTTCCTTGTTTTTGTCATTGCTGAGTGTTACATVFVIAECYMLTVMAYDRYVAIGCTGACAGTGATGGCCTACGACCGCTATGTTGCCATCTGCCACCCTTTGCTTTACAACHPLLYNIIMSHHTCLLLVAVCATCATTATGTCTCATCACACCTGCCTGCTGCTGGTGGCTGTGGTCTACGCCATCGGVYAIGLIGSTIETGLMLKLPYACTCATTGGCTCCACAATAGAAACTGGCCTCATGTTAAAACTGCCCTATTGTGAGCACEHLISHYFCDILPLMKLSCSCCTCATCAGTCACTACTTCTGTGACATCCTCCCTCTCATGAAGCTGTCCTGCTCTAGSTYDVEMTVFFSAGFNIIVTSCACCTATGATGTTGAGATGACAGTCTTCTTTTCGGCTGGATTCAACATCATAGTCACLTVLVSYTFILSSILGISTTEGAGCTTAACAGTTCTTGTTTCTTACACCTTCATTCTCTCCAGCATCCTCGGCATCAGGRSKAFSTCSSHLAAVGMFYGCACCACAGAGGGGAGATCCAAAGCCTTCAGCACCTGCAGCTCCCACCTTGCAGCCGTSTAFMYLKPSTISSLTQENVAGGGAATGTTCTATGGATCAACTGCATTCATGTACTTAAAACCCTCCACAATCAGTTCSVFYTTVIPMLNPLIYSLRNKCTTGACCCAGGAGAATGTGGCCTCTGTGTTCTACACCACGGTAATCCCCATGTTGAAEVKAAVQKTLRGKLFTCCCCTAATCTACAGCCTGAGGAACAAGGAAGTAAAGGCTGCCGTGCAGAAAACGCTGAGGGGTAAACTGTTTTGATGCAAATGMAP000818_E5/6AAAATGGTAAAAGGAAATCATTCCACGGTGACTGAATTTAATCTCGCTGGGCTAACAMVKGNHSTVTEFNLAGLTDKPGACAAACCAGAGCTCCAGCTGCCTCTTTTCCTCCTCTTCCTGGGAATCTATGTGGTCELQLPLFLLFLGIYVVTVVGNACAGTGGTGGGCAACCTGAGCATGATCACTCTAATAGGGTTCAGTTCTCACCTGCACLSMITLIGFSSHLHTPMYHFLACCCCCATGTACCATTTCCTCAGCAGTCTGTCCTTCATTGATCTCTGCCAGTCTTCTSSLSFIDLCQSSVITPKMLVNGTCATTACCCCAAAAATGCTGGTGAATTTTGTGTCAGAGAGGAATATTATCTCCTACFVSERVIISYPACMTQLYFFLCCAGCATGCATGACTCAGCTCTACTTCTTCCTTGTTCTTGTCATATCTGAATGTCACVLVISECHMLAAMAYDHYIAIATGTTGGCTGCAATGGCTTATGACCACTACATTGCCATATGTAACCCACTGCTTTACCMPLLYHVAMSYQVCSWMVVECATGTCGCCATGTCTTATCAGGTCTGCTCCTGGATGGTAGTTGAGGTGTATTTTATGVYFMGFIGATCSHSLHAKSAFGGCTTTATTGGTGCTACGTGCTCACACAGTCTGCATGCTAAGAGTGCTTTTCTGGAALEGRCNQPLLLGSFPTTGALPGGCCGATGTAATCAACCATTACTTCTGGGATCTTTTCCCACTACTGGAGCTCTCCCGLQYFYQRNSSLCFSAFNILFRCTCCAGTATTTCTATCAAAGAAATAGTAGTTTGTGCTTCAGTGCATTTAATATCCTTSLTILSSYIFIVASILCIRSTTTCCGCAGCCTCACCATCCTTAGCTCTTACATCTTCATCGTTGCCAGCATCCTCTGCEGRSKTFSTCSSHISAVSVFFATTCGCTCCACTGAGGGCAGGTCCAAAACCTTCAGCACTTGCAGCTCCCACATCTCGGSAAFMYLQPSSVSSMDQGSVGCTGTTTCTGTTTTCTTTGGGTCTGCAGCATTCATGTACCTGCAGCCATCATCCGTCFCVLCYCCAHAEPPIYSLRNKAGCTCCATGGACCAGGGGAGTGTCTTCTGTGTTTTATGCTACTGTTGTGCCCATGCTSVKVALIKFLEKRSFLGAACCCCCAATCTACAGCCTGAGGAATAAAGATGTCAAAGTTGCCTTAATTAAGTTCCTTGAAAAAAGAAGTTTCCTGTGAAAGMAP000818_A_17/8ATGGTGGACCCTGGAAACCATTCCTCAGTGACTGAGTCCATTCTGGCTGGGCTCTCAMVDPGNHSSVTESILAGLSEQGAACAGCCAGAGCTCCAGCTGCGCCTCTTCCTCCTGTTCTTAGGAATCTGTGTGGTCPELQLRLFLLFLGICVVTVVGACAGTGGTGGGCAACTTGGGCATGATCACACTGATTGGGCTCAGTTCTCACCTGCACNLGMITLIGLSSHLHTPMYYFACACCTATGTACTATTTCCTCAGCAGTCTGTCCTTCATTGACTTCTGCCATTCCACTLSSLSFIDFCHSTVITPKMLVGTCATTACCCCTAAGATGCTGGTGAACTTTGCGACAGAGAAGAACATCATCTCCTACNFATEKNIISYPECMAQLYLFCCTGAATGCATGGCTCAGCTCTATTTATTCAGTATTTTTGCTATTGCAGAGTGTCACSIFAIAECHMLAAMAYDCYVAATGTTGGCTGCAATGGCGTATGACTGTTATGTTGCCATCTGCAGCCCCTTGCTGTACECSPLLYNVIMSYHHCFWLTVAATGTCATCATGTCCTATCACCACTGCTTCTGGCTCACAGTGGGAGTTTACATTTTAGVYILGILGSTIHTSFMLRLFGGCATCCTTGGATCTACAATTCATACCAGTTTTATGTTGAGACTCTTTTTGTGCAAGLCKTNVINHYFCDLFPLLGLSACTAATGTGATTAACCATTATTTTTGTGATCTTTTCCCTCTCTTGGGGCTCTCCTGCCSSTYINELLVLVLSAFNILMTCCAGCACCTACATCAATGAATTACTGGTTCTGGTCTTGAGTGCATTTAACATCCTGPALTILASYIFIIASILRIHSATGCCTGCCTTAACCATCCTTGCTTCTTACATCTTTATCATTGCCAGCATCCTCCGCTEGRSKAFSTCSSHILAVAVFATTCACTCCACTGAGGGCAGGTCCAAAGCCTTCAGCACTTGCAGCTCCCACATCTTGFGSAAFMYLQPSSVSSMDQRKGCTGTTGCTGTTTTCTTTGGATCTGCAGCATTCATGTACCTGCAGCCATCATCTGTCVSSVFYTTIVPMLNPLIYSLRAGCTCCATGGACCAGAGGAAAGTGTCGTCTGTGTTTTATACTACTATTGTGCCCATGNKDVKLAVKKILHQTACCTGAACCCCCTGATCTACAGCCTGAGGAATAAAGATGTCAAACTTGCCGTGAAGAAAATTCTGCATCAGACAGCATGTTAATGAATAGAATCAATGTTATGMAP000818_C 9/10ATGGTGGACCCTGGAAACCATTCCTCAGTGACTGAGTCCATTCTGGCTGGGCTCTCAMVDPGNHSSVTESILAGLSEQGAACAGCCAGAGCTCCAGCTGCGCCTCTTCCTCCTGTTCTTAGGAATCTGTGTGGTCPELQLTLFLLFLGICVVTVVGACAGTGGTGGGCAACTTGGGCATGATCACACTGATTGGGCTCAGTTCTCACCTGCACNLGMITLIGLSSHLHTPMYYFACACCTATGTACTATTTCCTCAGCAGTCTGTCCTTCATTGACTTCTGCCATTCCACTLSSLSFIDFCHSTVITPKMLVGTCATTACCCCTAAGATGCTGGTGAACTTTGCGACAGAGAAGAACATCATCTCCTACNFATEKNIISYPECMAQLYLFCCTGAATGCATGGCTCAGCTCTATTTATTCAGTATTTTTGCTATTGCAGAGTGTCACSIFAIAECHMLAAMAYDCYVAATGTTGGCTGCAATGGCGTATGACTGTTATGTTGCCATCTGCAGCCCCTTGCTGTACICSPLLYNVIMSYHHCFWLTVAATGTCATCATGTCCTATCACCACTGCTTCTGGCTCACAGTGGGAGTTTACATTTTAGVYILGILGSTIHTSFMLRLFGGCATCCTTGGATCTACAATTCATACCAGTTTTATGTTGAGACTCTTTTTGTGCAAGLCKTNVINHYFCDLFPLLGLSACTAATGTGATTAACCATTATTTTTGTGATCTTTTCCCTCTCTTGGGGCTCTCCTGCCSSTYINELLVLVLSAFNILMTCCAGCACCTACATCAATGAATTACTGGTTCTGGTCTTGAGTGCATTTAACATCCTGPALTILASYIFIIASILRIHSATGCCTGCCTTAACCATCCTTGCTTCTTACATCTTTATCATTGCCAGCATCCTCCGCTEGRSKAFSTCSSHILAVAVFATTCACTCCACTGAGGGCAGGTCCAAAGCCTTCAGCACTTGCAGCTCCCACATCTTGFGSAAFMYLQPSSVSSMDQRKGCTGTTGCTGTTTTCTTTGGATCTGCAGCATTCATGTACCTGCAGCCATCATCTGTCVSSVFYTTIVPMLNPLIYSLRAGCTCCATGGACCAGAGGAAAGTGTCGTCTGTGTTTTATACTACTATTGTGCCCATGNKDVKLAVKKILHQTACCTGAACCCCCTGATCTACAGCCTGAGGAATAAAGATGTCAAACTTGCCGTGAAGAAAATTCTGCATCAGACAGCATGTTAATGAATAGAATCAATGTTATGMAP000818_F11/12ATGGTGGACCCTGGAAACCATTCCTCAGTGACTGAGTCCATTCTGGCTGGGCTCTCAMVDPGNHSSVTESILAGLSEQGAACAGCCAGAGCTCCAGCTGCGCCTCTTCCTCCTGTTCTTAGGAATCTGTGTGGTCPELQLRLFLLFLGICVVTVVGACAGTGGTGGGCAACTTGGGCATGATCACACTGATTGGGCTCAGTTCTCACCTGCACNLGMITLIGLSSHLHTPMYYFACACCTATGTACTATTTCCTCAGCAGTCTGTCCTTCATTGACTTCTGCCATTCCACTLSSLSFIDFCHSTVITPKMLVGTCATTACCCCTAAGATGCTGGTGAACTTTGCGACAGAGAAGAACATCATCTCCTACNFATEKNIISYPECMAQLYLFCCTGAATGCATGGCTCAGCTCTATTTATTCAGTATTTTTGCTATTGCAGAGTGTCACSIFAIAECHMLAAMAYDCYVAATGTTGGCTGCAATGGCGTATGACTGTTATGTTGCCATCTGCAGCCCCTTGCTGTACICSPLLYNVIMSYHHCFWLTVAATGTCATCATGTCCTATCACCACTGCTTCTGGCTCACAGTGGGAGTTTACATTTTAGVYILGILGSTIHTSFMLRLFGGCATCCTTGGATCTACAATTCATACCAGTTTTATGTTGAGACTCTTTTTGTGCAAGLCKTNVINHYFCDLFPLLGLSACTAATGTGATTAACCATTATTTTTGTGATCTTTTCCCTCTCTTGGGGCTCTCCTGCCSSTYINELLVLVLSAFNILMTCCAGCACCTACATCAATGAATTACTGGTTCTGGTCTTGAGTGCATTTAACATCCTGPALTILASYIFIIASILRIHSATGCCTGCCTTAACCATCCTTGCTTCTTACATCTTTATCATTGCCAGCATCCTCCGCTEGRSKAFSTCSSHILAVAVFATTCACTCCACTGAGGGCAGGTCCAAAGCCTTCAGCACTTGCAGCTCCCACATCTTGFGSAAFMYLQPSSVSSMDQRKGCTGTTGCTGTTTTCTTTGGATCTGCAGCATTCATGTACCTGCAGCCATCATCTGTCVSSVFYTTIVPMLNPLIYSLRAGCTCCATGGACCAGAGGAAAGTGTCGTCTGTGTTTTATACTACTATTGTGCCCATGNKDVKLAVKKILHQTACCTGAACCCCCTGATCTACAGCCTGAGGAATAAAGATGTCAAACTTGCCGTGAAGAAAATTCTGCATCAGACAGCATGTTAATGAATAGAATCAATGTTATCG55976-0113/14ATGGTGGACCCTGGAAACCATTCCTCAGTGACTGAGTCCATTCTGGCTGGGCTCTCAMVDPGNHSSVTESILAGLSEQGAACAGCCAGAGCTCCAGCTGCGCCTCTTCCTCCTGTTCTTAGGAATCTGTGTGGTCPELQLRLFLLFLGICVVTVVGACAGTGGTGGGCAACTTGGGCATGATCACACTGATTGGGCTCAGTTCTCACCTGCACNLGMITLIGLSSHLHTPMYYFACACCTATGTACTATTTCCTCAGCAGTCTGTCCTTCATTGACTTCTGCCATTCCACTLSSLSFIDFCHSTVITPKMLVGTCATTACCCCTAAGATGCTGGTGAACTTTGCGACAGAGAAGAACATCATCTCCTACNFATEKNIISYPECMAQLYLFCCTGAATGCATGGCTCAGCTCTATTTATTCAGTATTTTTGCTATTGCAGAGTGTCACSIFAIAECHMLAAMAYDCYVAATGTTGGCTGCAATGGCGTATGACTGTTATGTTGCCATCTGCAGCCCCTTGCTGTACICSPLLYNVIMSYHHCFWLTVAATGTCATCATGTCCTATCACCACTGCTTCTGGCTCACAGTGGGAGTTTACATTTTAGVYILGILGSTIHTSFMLRLFGGCATCCTTGGATCTACAATTCATACCAGTTTTATGTTGAGACTCTTTTTGTGCAAGLCKTNVINHYFCDLFPLLGLSACTAATGTGATTAACCATTATTTTTGTGATCTTTTCCCTCTCTTGGGGCTCTCCTGCCSSTYINELLVLVLSAFNILMTCCAGCACCTACATCAATGAATTACTGGTTCTGGTCTTGAGTGCATTTAACATCCTGPALTILASYIFIIASILRIHSATGCCTGCCTTAACCATCCTTGCTTCTTACATCTTTATCATTGCCAGCATCCTCCGCTEGRSKAFSTCSSHILAVAVFATTCACTCCACTGAGGGCAGGTCCAAAGCCTTCAGCACTTGCAGCTCCCACATCTTGFGSAAFMYLQPSSVSSMDQRKGCTGTTGCTGTTTTCTTTGGATCTGCAGCATTCATGTACCTGCAGCCATCATCTGTCVSSVFYTTIVPMLNPLIYSLRAGCTCCATGGACCAGAGGAAAGTGTCGTCTGTGTTTTATACTACTATTGTGCCCATGNKDVKLAVKKILHQTACCTGAACCCCCTGATCTACAGCCTGAGGAATAAAGATGTCAAACTTGCCGTGAAGAAAATTCTGCATCAGACAGCATGTTAATGAATAGAATCAATGTTATCG55976-0215/16ATGGTGGACCCTGGAAACCATTCCTCAGTGACTGAGTCCATTCTGGCTGGGCTCTCAMVDPGNHSSVTESILAGLSEQGAACAGCCAGAGCTCCAGCTGCGCCTCTTCCTCCTGTTCTTAGGAATCTGTGTGGTCPELQLRLFLLFLGICVVTVVGACAGTGGTGGGCAACTTGGGCATGATCACACTGATTGGGCTCAGTTCTCACCTGCACNLGMITLIGLSSHLHTPMYYFACACCTATGTACTATTTCCTCAGCAGTCTGTCCTTCATTGACTTCTGCCATTCCACTLSSLSFIDFCHSTVITPKMLVGTCATTACCCCTAAGATGCTGGTGAACTTTGCGACAGAGAAGAACATCATCTCCTACNFATIKNIISYPECMAQLYLFCCTGAATGCATGGCTCAGCTCTATTTATTCAGTATTTTTGCTATTGCAGAGTGTCACSIFAIAECHMLAAMAYDCYVAATGTTGGCTGCAATGGCGTATGACTGTTATGTTGCCATCTGCAGCCCCTTGCTGTACICSPLLYNVIMSYHHCFWLTVAATGTCATCATGTCCTATCACCACTGCTTCTGGCTCACAGTGGGAGTTTACATTTTAGVYILGILGSTIHTSFMLRLFGGCATCCTTGGATCTACAATTCATACCAGTTTTATGTTGAGACTCTTTTTGTGCAAGLCKTNVINHYFCDLFPLLGLSACTAATGTGATTAACCATTATTTTTGTGATCTTTTCCCTCTCTTGGGGCTCTCCTGCCSSTYINELLVLVLSAFNILMTCCAGCACCTACATCAATGAATTACTGGTTCTGGTCTTGAGTGCATTTAACATCCTGPALTILASYIFIIASILRIHSATGCCTGCCTTAACCATCCTTGCTTCTTACATCTTTATCATTGCCAGCATCCTCCGCTEGRSKAFSTCSSHILAVAVFATTCACTCCACTGAGGGCAGGTCCAAAGCCTTCAGCACTTGCAGCTCCCACATCTTGFGSAAFMYLQPSSVSSMDQRKGCTGTTGCTGTTTTCTTTGGATCTGCAGCATTCATGTACCTGCAGCCATCATCTGTCVSSVFYTTIVPMLNPLIYSLRAGCTCCATGGACCAGAGGAAAGTGTCGTCTGTGTTTTATACTACTATTGTGCCCATGNKDVKLAVKKILHQTACCTGAACCCCCTGATCTACAGCCTGAGGAATAAAGATGTCAAACTTGCCGTGAAGAAAATTCTGCATCAGACAGCATGTTAATGAATAGAATCAATGTTATGMAC024257_A17/18ATGCTAAGGAATGGCAGCATAGTGACGGAATTTATCCTCGTGGGCTTTCAGCAGAGCMLRNGSIVTEFILVGFQQSSTTCCACTTCCACACGAGCATTGCTCTTTGCCCTCTTCTTGGCCCTCTACAGCCTCACCSTRALLFALFLALYSLTMAMNATGGCCATGAATGGCCTCATCATCTTTATCACCTCCTGGACAGACCCCAAGCTCAACGLIIFITSWTDPKLNSPMYFFAGCCCCATGTACTTCTTCCTCGGCCATCTGTCTCTCCTGGATGTCTGCTTCATCACCLGHLSLLDVCFITTTIPQMLIACTACCATCCCACAGATGTTGATCCACCTCGTGGTCAGGGACCACATTGTCTCCTTTHLVVRDHIVSFVCCMTQMYFVGTATGTTGCATGACCCAGATGTACTTTGTCTTCTGTGTTGGTGTGGCCGAGTGCATCFCVGVAECILLAFMAYDRYVACTCTTGGCTTTCATGGCCTATGACCGTTATGTTGCTATCTGCTACCCACTTAACTATICYPLNYVPIISQKVCVRLVGGTCCCGATCATAAGCCAGAAGGTCTGTGTCAGGCTTGTGGGAACTGCCTGGTTCTTTTAWFFGLINGIFLEYISFREPGGGCTGATCAATGGCATCTTTCTCGAGTATATTTCATTCCGAGAGCCCTTCCGCAGAFRRDNHIESFFCEAPIVIGLSGACAACCACATAGAAAGCTTCTTCTGTGAGGCCCCCATAGTGATTGGCCTCTCTTGTCGDPQFSLWAIFADAIVVILSGGGGACCCTCAGTTTAGTCTGTGGGCAATCTTTGCCGATGCCATCGTGGTAATTCTCPMVLTVTSYVHILATILSKASAGCCCCATGGTGCTCACTGTCACTTCCTATGTGCACATCCTGGCCACCATCCTCAGCSSGRGKTFSTCASHLTVVIFLAAAGCCTCCTCCTCAGGTCGGGGGAAGACTTTCTCTACTTGTGCCTCTCACCTGACTYTSAMFSYMNPHSTHGPDKDKGTGGTCATCTTTCTCTACACTTCAGCTATGTTCTCTTACATGAACCCCCACAGCACAPFSLLYTIITPMCNPIIYSFRCATGGGCCTGACAAAGACAAACCTTTCTCCCTCCTGTACACCATCATTACCCCCATGNKEIKEAMVRALGRTRLAQPQTGCAACCCCATCATTTATAGTTTCCGCAACAAGGAAATTAAGGAGGCCATGGTGAGGSVGCACTTGGAAGAACCAGGCTGGCCCAGCCACAGTCTGTCTAGCAGGAAGGCCCTGAAAGGAAAGCTCCTTCACCTGGGGAATGGGAGAGMAC011537_A19/20GGGTGGCCGCCATGCAGGGGCTAAACCACACCTCCGTGTCTGAATTCATCCTCGTTGMQGLNHTSVSEFILVGFSAFPGCTTCTCTGCCTTCCCCCACCTCCAGCTGATGCTCTTCCTGCTGTTCCTGCTGATGTHLQLMLFLLFLLMYLFTLLGNACCTGTTCACGCTGCTGGGCAACCTGCTCATCATGGCCACTGTCTGGAGCGAGCGCALLIMATVWSERSLHMPMYLFLGCCTCCACATGCCCATGTACCTCTTCCTGTGTGCCCTCTCCATCACCGAGATCCTCTCALSITEILYTVAIIPRMLADACACCGTGGCCATCATCCCGCGCATGCTGGCCGACCTGCTGTCCACCCAGCGCTCCALLSTQRSIAFLACASQMFFSFTCGCCTTCCTGGCCTGTGCCAGTCAGATGTTCTTCTCCTTCAGCTTCGGCTTCACCCSFGFTHSFLLTVMGYDRYVAIACTCCTTCCTGCTCACTGTCATGGGCTACGACCGCTACGTGGCCATCTGCCACCCCCCHPLRYNVLMSLRGCTCRVGCTGCGTTACAACGTGCTCATGAGCCTGCGGGGCTGCACCTGCCGGGTGGGCTGCTCCTSWAGGLVMGMVVTSAIFHLAFGGGCTGGTGGCTTGGTCATGGGGATGGTGGTGACCTCGGCCATTTTCCACCTCGCCTCGHKEIHHFFCHVPPLLKLACTCTGTGGACACAAGGAGATCCACCATTTCTTCTGCCACGTGCCACCTCTGTTGAAGTGDDVLVVAKGVGLVCITALLGTGGCCTGTGGAGATGATGTGCTGGTGGTGGCCAAAGGCGTGGGCTTGGTGTGTATCACFLLILLSYAFIVAAILKIPSCGGCCCTGCTGGGCTGTTTTCTCCTCATCCTCCTCTCCTATGCCTTCATCGTGGCCGAEGRNKAFSTCASHLTVVVVHCCATCTTGAAGATCCCTTCTGCTGAAGGTCGGAACAAGGCCTTCTCCACCTGTGCCTYGFASVIYLKPKGPQSPEGDTCTCACCTCACTGTGGTGGTCGTGCACTATGGCTTTGCCTCCGTCATTTACCTGAAGCLMGITYTVLTPFLSPIIFSLRCCAAAGGTCCCCAGTCTCCGGAAGGAGACACCTTGATGGGCATCACCTACACGGTCCNKELKVAMKKTCFTKLFPQNCTCACACCCTTCCTCAGCCCCATCATCTTCAGCCTCAGGAACAAGGAGCTGAAGGTCGCCATGAAGAAGACTTGCTTCACCAAACTCTTTCCACAGAACTGCTGAAATGGCTGACTTTCTCTCAAGAGATGMAP001804_E21/22AATGACTCTGAGAAACAGCTCCTCAGTGACTGAGTTTATCCTTGTGGGATTATCAGAMTLRNSSSVTEFILVGLSEQPACAGCCAGAGCTCCAGCTCCCTCTTTTCCTTCTATTCTTAGGGATCTATGTGTTCACELQLPLFLLFLGIYVFTVVGNTGTGGTGGGCAACTTGGGCTTGATCACCTTAATTGGGATAAATCCTAGCCTTCACACLGLITLIGINPSLHTPMYFFLCCCCATGTACTTTTTCCTCTTCAACTTGTCCTTTATAGATCTCTGTTATTCCTGTGTFNLSFIDLCYSCVFTPKMLNDGTTTACCCCCAAAATGCTGAATGACTTTGTTTCAGAAAGTATCATCTCTTATGTGGGFVSESIISYVGCMTQLFFFCFATGTATGACTCAGCTATTTTTCTTCTGTTTCTTTGTCAATTCTGAGTGCTATGTGTTFVNSECYVLVSMAYDRYVAICGGTATCAATGGCCTATGATCGCTATGTGGCCATCTGCAACCCCCTGCTCTACATGGTNPLLYMVTMSPRVCFLLMFGSCACCATGTCCCCAAGGGTCTGCTTTCTGCTGATGTTTGGTTCCTATGTGGTAGGGTTYVVGFAGAMAHTGSMLRLTFCTGCTGGGGCCATGGCCCACACTGGAAGCATGCTGCGACTGACCTTCTGTGATTCCAADSNVIDHYLCDVLPLLQLSCTCGTCATTGACCATTATCTGTGTGACGTTCTCCCCCTCTTGCAGCTCTCCTGCACCAGSTHVSELVFFIVVGVITMLSSCACCCATGTCAGTGAGCTGGTATTTTTCATTGTTGTTGGAGTAATCACCATGCTATCISIVISYALILSNILCIPSAECAGCATAAGCATCGTCATCTCTTACGCTTTGATACTCTCCAACATCCTCTGTATTCCGRSKAFSTWGSHIIAVALFFGTTCTGCAGAGGGCAGATCCAAAGCCTTTAGCACATGGGGCTCCCACATAATTGCTGTSGTFTYLTTSFPGSMNHGRFATGCTCTGTTTTTTGGGTCAGGGACATTCACCTACTTAACAACATCTTTTCCTGGCTCSVFYTNVVPMLNPSIYSLRNKTATGAACCATGGCAGATTTGCCTCAGTCTTTTACACCAATGTGGTTCCCATGCTTAADDKLALGKTLKRVLFCCCTTCGATCTACAGTTTGAGGAATAAGGATGATAAACTTGCCCTGGGCAAAACCCTGAAGAGAGTGCTCTTCTAATGGGMAP001804_A23/24AAGAATGACCATGGAAAATTATTCTATGGCAGCTCAGTTTGTCTTAGATGGMTMENYSMAAQFVLDGLTTTTAACACAGCAAGCAGAGCTCCAGCTGCCCCTCTTCCTCCTGTTCCTGGGQQAELQLPLFLLFLGIYVVTAATCTATGTGGTCACAGTAGTGGGCAACCTGGGCATGATTCTCCTGATTGCVVGNLGMILLIAVSPLLHTPAGTCAGCCCTCTACTTCACACCCCCATGTACTATTTCCTCAGCAGCTTGTCCMYYFLSSLSFVDFCYSSVITTTCGTCGATTTCTGCTATTCCTCTGTCATTACTCCCAAAATGCTGGTGAACTPLMLVNFLGKKNTILYSECTCCTAGGAAAGAAGAATACAATCCTTTACTCTGAGTGCATGGTCCAGCTCTMVQLFFFVVFVVAEGYLLTTTTTCTTTGTGGTCTTTGTGGTGGCTGAGGGTTACCTCCTGACTGCCATGGCAMAYDRYVAICSPLLYNAIATATGATCGCTATGTTGCCATCTGTAGCCCACTGCTTTATAATGCGATCATMSSWVCSLLVLAAFFLGFLGTCCTCATGGGTCTGCTCACTGCTAGTGCTGGCTGCCTTCTTCTTGGGCTTTSALTHTSAMMKLSFCKSHIICTCTCTGCCTTGACTCATACAAGTGCCATGATGAAACTGTCCTTTTGCAAATNHYFCDVLPLLNLSCSNTHCCCACATTATCAACCATTACTTCTGTGATGTTCTTCCCCTCCTCAATCTCTCLNELLLFIIAGFNTLVPTLACTGCTCCAACACACACCTCAATGAGCTTCTACTTTTTATCATTGCGGGGTTTVAVSYAFILYSILHIRSSEGRAACACCTTGGTGCCCACCCTAGCTGTTGCTGTCTCCTATGCCTTCATCCTCTSKAFGTCSSHLMAVVIFFGACAGCATCCTTCACATCCGCTCCTCAGAGGGCCGGTCCAAAGCTTTTGGAASITFMYFKPPSSNSLDQEKVCATGCAGCTCTCATCTCATGGCTGTGGTGATCTTCTTTGGGTCCATTACCTTSSVFYTTVIPMLNPLIYSLRCATGTATTTCAAGCCCCCTTCAAGTAACTCCCTGGACCAGGAGAAGGTGTCNKDVKKALRKVLVGKCTCTGTGTTCTACACCACGGTGATCCCCATGCTGAACCCTTTAATATACAGTGMAC011711_G25/26GGCACCCACCTTCCCCCTTGTCTCCTCACACAATGACCCTGGGATCCCTGGGAAACAMTLGSLGNSSSVSATFLLSGGCAGCAGCAGCGTTTCTGCTACCTTCCTGCTGAGTGGCATCCCTGGGCTGGAGCGCAIPGLERMHIWISIPLCFMYLVTGCACATCTGGATCTCCATCCCACTGTGCTTCATGTATCTGGTTTCCATCCCGGGCASIPGNCTILFIIKTERSLHEPACTGCACAATTCTTTTTATCATTAAAACAGAGCGCTCACTTCATGAACCTATGTATCMYLFLSMLALIDLGLSLCTLPTCTTCCTGTCCATGCTGGCTCTGATTGACCTGGGTCTCTCCCTTTGCACTCTCCCTATVLGIFWVGAREISHDACFAQCAGTCCTGGGCATCTTTTGGGTTGGAGCACGAGAAATTAGCCATGATGCCTGCTTTGLFFIHCFSFLESSVLLSMAFDCTCAGCTCTTTTTCATTCACTGCTTCTCCTTCCTCGAGTCCTCTGTGCTACTGTCTARFVAICHPLHYVSILTNTVIGTGGCCTTTGACCGCTTTGTGGCTATCTGCCACCCCTTGCACTATGTTTCCATTCTCARIGLVSLGRSVALIFPLPFMLCCAACACAGTCATTGGCAGGATTGGCCTGGTCTCTCTGGGTCGTAGTGTAGCACTCAKRFPYCGSPVLSHSYCLHQEVTTTTTCCATTACCTTTTATGCTCAAAAGATTCCCCTATTGTGGCTCCCCAGTTCTCTMKLACADMKANSIYGMFVIVSCACATTCTTATTGTCTCCACCAAGAAGTGATGAAATTGGCCTGTGCCGACATGAAGGTVGIDSLLILFSYALILRTVLCCAACAGCATCTACGGCATGTTTGTCATCGTCTCTACAGTGGGTATAGACTCACTGCSIASRAERFKALNTCVSHICATCATCCTCTTCTCTTATGCTCTGATCCTGCGCACCGTGCTGTCCATCGCCTCCAGGGVLLFYTPMIGLSVIHRFGKQACTGAGAGATTCAAGGCCCTTAACACCTGTGTTTCCCACATCTGTGCTGTGCTGCTCTPHLVQVVMGFMYLLFPPVMNPTCTACACTCCCATGATTGGCCTCTCTGTCATCCATCGCTTTGGAAAGCAGGCACCCCIVYSVKTKQIRDRVTHAFCYACCTGGTCCAGGTGGTCATGGGTTTCATGTATCTTCTCTTTCCTCCTGTGATGAATCCCATTGTCTACAGTGTGAAGACCAAACAGATCCGGGATCGAGTGACGCATGCCTTTTGTTACTAACTGMAC011711_F27/28TGCTTCCCTATGTCGGTCCTCAATAATACCATTGCTGAGCCTCTGATCTTCCTCCTGMSVLNNTIAEPLIFLLMGIPGATGGGCATTCCAGGCCTGAAAGCCACCCAGTACTGGATCTCCATCCCTTTTTGTCTCLKATQYWISIPFCLLYVVAVSCTATATGTTGTTGCCGTCTCTGGAAATAGCATGATCCTGTTTGTGGTCCTCTGTGAAGNSMILFVVLCERSLHKPMYYCGGAGCCTCCATAAGCCTATGTACTATTTCCTCTCTATGCTTTCAGCCACAGACCTGFLSMLSATDLSLSLCTLSTTLAGCTTGTCCCTGTGTACACTTTCTACTACCCTTGGTGTCTTCTGGTTTGAAGCCCGAGVFWFEAREINLNACIAQMFFGAAATCAACCTAAATGCCTGCATTGCCCAGATGTTCTTTCTACACGGATTTACTTTCLHGFTFMESGVLLAMAFDRFVATGGAGTCTGGGGTTCTACTGGCCATGGCCTTTGATCGTTTTGTGGCCATCTGTTACAICYPLRYTTILTNARIAKIGCCACTGAGATACACTACCATCCTTACCAATGCCCGAATTGCCAAGATTGGGATGAGCMSMLIRNVAVMLPVMLFVKRLATGTTGATAAGAAATGTTGCCGTCATGTTGCCAGTCATGCTCTTTGTCAAGAGGTTGSFCSSMVLSHSYCYGVDLIQLTCCTTCTGCAGTTCTATGGTCCTTTCACATTCTTACTGCTACCATGTTGATCTCATCSCTDNRINSILGLFALLSTTGCAACTCTCCTGCACAGACAATAGGATCAACAGCATCCTTGGTCTGTTTGCGCTTTTGFDCPCILLSYILIIRSVLSIATCCACTACAGGGTTTGACTGCCCTTGCATCCTGCTCTCCTATATCCTGATCATTCGASSEERRKAFNTCTSHISAVSITCTGTCCTCAGCATTGCTTCCTCAGAAGAGAGGCGGAAAGCCTTCAACACCTGCACAFYLPLISLSLVHRYGHSAPPFTCCCACATCAGTGCTGTTTCCATCTTCTACCTCCCTCTCATCAGTTTGTCTCTTGTCVHIIMANVFLLIPPVLNPIIYCATCGCTATGGCCATTCAGCACCTCCATTTGTCCACATCATCATGGCCAATGTCTTTSVKIKQIQKAIIKVLIQKHSKCTGCTAATCCCTCCTGTGCTCAACCCTATTATTTACAGTGTAAAGATTAAGCAGATTSNHQLFLIRDKAIYECAAAAGGCCATTATCAAGGTCTTAATTCAGAAGCACTCCAAATCTAATCATCAGCTATTTCTGATTAGAGATAAAGCCATTTATGAATAACG55978-0129/30TGCTTCCCTATGTCGGTCCTCAATAATACCATTGCTGAGCCTCTGATCTTCCTCCTGMSVLNNTIAEPLIFLLMGIPGATGGGCATTCCAGGCCTGAAAGCCACCCAGTACTGGATCTCCATCCCTTTTTGTCTCLKATQYWISIPFCLLYVVAVSCTATATGTTGTTGCCGTCTCTGGAAATAGCATGATCCTGTTTGTGGTCCTCTGTGAAGNSMILFVVLCERSLHKPMYYCGGAGCCTCCATAAGCCTATGTACTATTTCCTCTCTATGCTTTCAGCCACAGACCTGFLSMLSATDLSLSLCTLSTTLAGCTTGTCCCTGTGTACACTTTCTACTACCCTTGGTGTCTTCTGGTTTGAAGCCCGAGVFWFEAREINLNACIAQMFFGAAATCAACCTAAATGCCTGCATTGCCCAGATGTTCTTTCTACACGGATTTACTTTCLHGFTFMESGVLLAMAFDRFVATGGAGTCTGGGGTTCTACTGGCCATGGCCTTTGATCGTTTTGTGGCCATCTGTTACAICYPLRYTTILTNARIAKIGCCACTGAGATACACTACCATCCTTACCAATGCCCGAATTGCCAAGATTGGGATGAGCMSMLIRNVAVMLPVMLFVKRLATGTTGATAAGAAATGTTGCCGTCATGTTGCCAGTCATGCTCTTTGTCAAGAGGTTGSFCSSMVLSHSYCYHVDLIQLTCCTTCTGCAGTTCTATGGTCCTTTCACATTCTTACTGCTACCATGTTGATCTCATCSCTDNRINSILGLFALLSTTGCAACTCTCCTGCACAGACAATAGGATCAACAGCATCCTTGGTCTGTTTGCGCTTTTGFDCPCILLSYILIIRSVLSIATCCACTACAGGGTTTGACTGCCCTTGCATCCTGCTCTCCTATATCCTGATCATTCGASSEERRKAFNTCTSHISAVSITCTGTCCTCAGCATTGCTTCCTCAGAAGAGAGGCGGAAAGCCTTCAACACCTGCACAFYLPLISLSLVHRYGHSAPPFTCCCACATCAGTGCTGTTTCCATCTTCTACCTCCCTCTCATCAGTTTGTCTCTTGTCVHIIMANVFLLIPPVLNPIIYCATCGCTATGGCCATTCAGCACCTCCATTTGTCCACATCATCATGGCCAATGTCTTTSVKIKQIQKAIIKVLIQKHSKCTGCTAATCCCTCCTGTGCTCAACCCTATTATTTACAGTGTAAAGATTAAGCAGATTSNHQLFLIRDKAIYECAAAAGGCCATTATCAAGGTCTTAATTCAGAAGCACTCCAAATCTAATCATCAGCTATTTCTGATTAGAGATAAAGCCATTTATGAATAAGMAC011711_E31/32TTCAACGTGGAATCCTGAACTATGTCAACATTACCAACTCAGATAGCCCCCAATAGCMSTLPTQIAPNSSTSMAPTFLAGCACTTCAATGGCCCCCACCTTCTTGCTGGTGGGCATGCCAGGCCTATCAGGTGCALVGMPGLSGAPSWWTLPLIAVCCCTCCTGGTGGACATTGCCCCTCATTGCTGTCTACCTTCTCTCTGCACTGGGAAATYLLSALGNGTILWIIALQPALGGCACCATCCTCTGGATCATTGCCCTGCAGCCCGCCCTGCACCGCCCAATGCACTTCHRPMHFFLFLLSVSDIGLVTATTCCTCTTCTTGCTTAGTGTGTCTGATATTGGATTGGTCACTGCCCTGATGCCCACALMPTLLGIALAGAHTVPASACCTGCTGGGCATCGCCCTTGCTGGTGCTCACACTGTCCCTGCCTCAGCCTGCCTTCTALLQMVFIHVFSVMESSVLLAMCAGATGGTTTTTATCCATGTCTTTTCTGTCATGGAGTCCTCTGTCTTGCTCGCCATGSIDRALAICRPLHYPALLTNGTCCATTGATCGGGCACTGGCCATCTGCCGACCTCTCCACTACCCAGCGCTCCTCACCVISKISLAISFRCLGLHLPLPAATGGTGTAATTAGCAAAATCAGCCTGGCCATTTCTTTTCGATGCCTGGGTCTCCATFLLAYMPYCLPQVLTHSYCLHCTGCCCCTGCCATTCCTGCTGGCCTACATGCCCTACTGCCTCCCACAGGTCCTAACCPDVARLACPEAWGAAYSLFVVCATTCTTATTGCTTGCATCCAGATGTGGCTCGTTTGGCCTGCCCAGAAGCTTGGGGTLSAMGLDPLLIFFSYGLIGKVGCAGCCTACAGCCTATTTGTGGTTCTTTCAGCCATGGGTTTGGACCCCCTGCTTATTLQGVESREDRWKAGQTCAAHLTTCTTCTCCTATGGCCTGATTGGCAAGGTGTTGCAAGGTGTGGAGTCCAGAGAGGATSAVLLFYIPMILLALINHPELCGCTGGAAGGCTGGTCAAACCTGTGCTGCCCACCTCTCTGCAGTGCTCCTCTTCTATPITQHTHTLLSYVHFLLPPLIATCCCTATGATCCTCCTGGCACTGATTAACCATCCTGAGCTGCCAATCACTCAGCATNPILYSVKMKEIRKRILNRLQACCCATACTCTTCTATCCTATGTCCATTTCCTTCTTCCTCCATTGATAAACCCTATTPRKVGGAQCTCTATAGTGTCAAGATGAAGGAGATTAGAAAGAGAATACTCAACAGGTTGCAGCCCAGGAAGGTGGGTGGTGCTCAGTGAGMAC011711_D33/34CTATGACAATTCTTCTTAATAGCAGCCTCCAAAGAGCCACTTTCTTCCTGACGGGCTMTILLNSSLQRATFFLTGFQGTCCAAGGTCTAGAAGGTCTCCATGGCTGGATCTCTATTCCCTTCTGCTTCATCTACCLEGLHGWISIPFCFIYLTVILTGACAGTTATCTTGGGGAACCTCACCATTCTCCACGTCATTTGTACTGATGCCACTCGNLTILHVICTDATLHGPMYYTCCATGGACCCATGTACTATTTCTTGGGCATGCTAGCTGTCACAGACTTAGGCCTTTFLGMLAVTDLGLCLSTLPTVLGCCTTTCCACACTGCCCACTGTGCTGGGCATTTTCTGGTTTGATACCAGAGAGATTGGIFWFDTREIGIPACFTQLFFGCATCCCTGCCTGTTTCACTCAGCTCTTCTTCATCCACACCTTGTCTTCAATGGAGTIHTLSSMESSVLLSMSIDRSVCATCAGTTCTGTTATCCATGTCCATTGACCGCTCCGTGGCCGTCTGCAACCCACTGCAVCNPLHDSTVLTPACIVKMGATGACTCCACCGTCCTGACACCTGCATGTATTGTCAAGATGGGGCTAAGCTCAGTGCLSSVLRSALLILPLPFLLKRFTTAGAAGTGCTCTCCTCATCCTCCCCTTGCCATTCCTCCTGAAGCGCTTCCAATACTQYCHSHVLAHAYCLHLEIMKLGCCACTCCCATGTGCTGGCTCATGCTTATTGTCTTCACCTGGAGATCATGAAGCTGGACSSIIVNHIYGLFVVACTVGCCTGCTCTAGCATCATTGTCAATCACATCTATGGGCTCTTTGTTGTGGCCTGCACCGVDSLLIFLSYALILRTVLSIATGGGTGTGGACTCCCTGCTCATCTTTCTCTCATACGCCCTCATCCTTCGCACCGTGCSHQERLRALNTCVSHICAVLLTCAGCATTGCCTCCCACCAGGAGCGACTCCGAGCCCTCAACACCTGTGTCTCTCATAFYIPMIGLSLVHRFGEHLPRVTCTGTGCTGTACTGCTCTTCTACATCCCCATGATTGGCTTGTCTCTTGTGCATCGCTVHLFMSYVYLLVPPLMNPIIYTTGGTGAACATCTGCCCCGCGTTGTACACCTCTTCATGTCCTATGTGTATCTGCTGGSIKTKQIRQRIIKKFQFIKSLTACCACCCCTTATGAACCCCATCATCTACAGCATCAAGACCAAGCAAATTCGCCAGCRCFWKDGCATCATTAAGAAGTTTCAGTTTATAAAGTCACTTAGGTGTTTTTGGATTAAGMAC011711_B35/36TCCATGGTGCTGGCTTCAGGGAACAGCTCTTCTCATCCTGTGTCCTTCATCCTGCTTMVLASGNSSSHPVSFILLGIPGGAATCCCAGGCCTGGAGAGTTTCCAGTTGTGGATTGCCTTTCCGTTCTGTGCCACGGLESFQLWIAFPFCATYAVAVTATGCTGTGGCTGTTGTTGGAAATATCACTCTCCTCCATGTAATCAGAATTGACCACVGNITLLHVIRIDHTLHEPMYACCCTGCATGAGCCCATGTACCTCTTTCTGGCCATGCTGGCCATCACTGACCTGGTCLFLAMLAITDLVLSSSTQPKMCTCTCCTCCTCCACTCAACCTAAGATGTTGGCCATATTCTGGTTTCATGCTCATGAGLAIFWFHAHEIQYHACLIQVFATTCAGTACCATGCCTGCCTCATCCAGGTGTTCTTCATCCATGCCTTTTCTTCTGTGFIHAFSSVESGVLMAMALDCYGAGTCTGGGGTGCTCATGGCTATGGCCCTGGACTGCTACGTGGCTACCTGCTTCCCAVATCFPLRHSSILTPSVVIKLCTCCGACACTCTAGCATCCTGACCCCATCGGTCGTGATCAAACTGGGGACCATCGTGGTIVMLRGLLWVSPFCFMVSRATGCTGAGAGGGCTGCTGTGGGTGAGCCCCTTCTGCTTCATGGTGTCTAGGATGCCCMPFCQHQAIPQSYCEHMAKLKTTCTGCCAACACCAAGCCATTCCCCAGTCATACTGTGAGCACATGGCTGTGCTGAAGLVCADTSISRGYGLFVAFSVATTGGTGTGTGCTGATACAAGCATAAGTCGTGGGTATGGGCTCTTTGTGGCCTTCTCTGFDMIVIGMSYVMILRAVLQLGTGGCTGGCTTTGATATGATTGTCATTGGTATGTCATACGTGATGATTTTGAGAGCTPSGEARLKAFSTRASHICVILGTGCTTCAGTTGCCCTCAGGTGAAGCCCGCCTCAAAGCTTTTAGCACACGTGCCTCCALYIPALFSFLTYRFGHDVPRCATATCTGTGTCATCTTGGCTCTTTATATCCCAGCCCTTTTTTCTTTCCTCACCTACVVHILFANLYLLIPPMLNPIICGCTTTGGCCATGATGTGCCCCGAGTTGTACACATCCTGTTTGCTAATCTCTATCTAYGVRTKQIGDRVIQGCCGNIPCTGATACCTCCCATGCTCAACCCCATCATTTATGGAGTTAGAACCAAACAGATCGGGGACAGGGTTATCCAAGGATGTTGTGGAAACATCCCCTGAGCAAAGGGMAC009758_B37/38CTATGCCTACTGTAAACCACAGTGGCACTAGCCACACAGTCTTCCACTTGCTGGGCAMPTVNHSGTSHTVFHLLGIPGTCCCTGGCCTACAGGACCAGCACATGTGGATTTCTATCCCATTCTTCATTTCCTATGLQDQHMWISIPFFISYVTALLTCACCGCCCTTCTTGGGAACAGCCTGCTCATCTTCATTATCCTCACAAAGCGCAGCCGNSLLIFIILTKRSLHEPMYLTCCATGAACCCATGTACCTCTTCCTCTGCATGCTGGCTGGAGCAGACATTGTCCTCTFLCMLAGADIVLSTCTIPQALCCACGTGCACCATTCCTCAGGCCTTAGCTATCTTCTGGTTCCGTGCTGGGGACATCTAIFWFRAGDISLDRCITQLFFCCCTGGATCGTTGCATCACTCAGCTCTTCTTCATCCATTCCACCTTCATCTCTGAGTIHSTFISESGILLVMAFDHYICAGGGATCTTGCTGGTGATGGCCTTTGACCACTATATTGCCATATGCTACCCACTGAAICYPLRYTTILTNALIKKICGGTACACCACCATTCTTACAAATGCTCTGATCAAGAAAATTTGTGTGACTGTCTCTCVTVSLRSYGTIFPIIFLLKRLTGAGAAGTTATGGTACAATTTTCCCTATCATATTTCTTTTAAAAAGATTGACTTTCTTFCQNNIIPHTFCEHIGLAKYGCCAGAATAATATTATTCCACACACCTTTTGTGAACACATTGGCCTAGCCAAATATGACNDIRINIWYGFSILMSTVVCATGTAATGACATTCGAATAAACATTTGGTATGGGTTTTCCATTCTAATGTCGACGGLDVVLIFISYMLILHAVFHMPTGGTCTTAGATGTTGTACTAATTTTTATTTCCTATATGCTGATTCTCCATGCTGTCTSPDACHKALNTFGSHVCIIILTCCACATGCCTTCTCCAGATGCTTGCCACAAAGCTCTCAACACATTTGGCTCCCATGFYGSGIFTILTQRFGRHIPPCTCTGCATCATCATCCTCTTTTATGGGTCTGGCATCTTCACAATCCTTACCCAGAGGTIHIPLANVCILAPPMLNPIIYTTGGACGCCACATTCCACCTTGTATCCACATCCCGTTGGCTAATGTCTGCATTCTGGGIKTKQIQCTCCACCTATGCTGAATCCCATTATTTATGGGATCAAAACCAAGCAAATCCAGTAACAGGTGGMAC023106_C39/40ATTCTTTTCCTCCCTGGATGTTTCTTTGCAGGAACTCATGCAGCCAGAATCTGGGGCMQPESGANGTVIAEFILLGLLCAATGGAACAGTCATTGCTGAGTTCATCCTGCTGGGCTTGCTGGAGGCGCCAGGGCTEAPGLQPVVFVLFLFAYLVTVGCAGCCAGTTGTCTTTGTGCTCTTCCTCTTTGCCTACCTGGTCACGGTCAGGGGCAARGNLSILAAVLVEPKLHTPMYCCTCAGCATCCTGGCAGCTGTCTTGGTGGAGCCCAAACTCCACACCCCCATGTACTTFFLGNLSVLDVGCISVTVPSMCTTCCTGGGGAACCTATCAGTGCTGGATGTTGGGTGCATCAGCGTCACTGTTCCATCLSRLLSRKRAVPCGACLTQLFAATGTTGAGTCGTCTCCTGTCCCGCAAGCGTGCAGTTCCCTGTGGGGCCTGCCTTACFFHLFVGVDCFLLTAMAYDRFCCAGCTCTTCTTCTTCCATCTGTTCGTTGGAGTGGACTGCTTCCTGCTGACCGCCATLAICRPLTYSTRMSQTVQRMLGGCCTATGACCGATTCCTGGCCATCTGCCGGCCCCTCACCTACAGCACCCGCATGAGVAASWACAFTNALTHTVAMSTTCAGACAGTCCAGAGGATGTTGGTGGCTGCGTCCTGGGCTTGTGCTTTCACCAACGCLNFCGPNVINHFYCDLPQLFQACTGACCCACACTGTGGCCATGTCCACGCTCAACTTCTGTGGCCCCAATGTGATCAALSCSSTQLNELLLFAVGFIMATCACTTCTACTGTGACCTCCCACAGCTCTTCCAGCTCTCCTGCTCCAGCACCCAACTGTPMALIVISYIHVAAAVLRICAATGAGCTGCTGCTTTTTGCTGTGGGTTTTATAATGGCAGGTACCCCCATGGCTCTRSVEGRKKAFSTCGSHLTVVACATTGTCATCTCCTATATCCACGTGGCAGCTGCAGTCCTGCGAATTCGCTCTGTAGAIFYGSGIFNYMRLGSTKLSDKGGGCAGGAAGAAAGCCTTCTCCACATGTGGCTCCCACCTCACTGTGGTTGCCATATTDKAVGIFNTVINPMLNPIIYSCTATGGTTCAGGTATCTTTAACTATATGCGACTGGGTTCAACCAAGCTTTCAGACAAFRNPDVQSAIWRMLTGRRSLAGGATAAAGCTGTTGGAATTTTCAACACTGTCATCAATCCCATGCTGAACCCAATCATCTACAGCTTCAGAAACCCTGATGTGCAGAGTGCCATCTGGAGGATGCTCACAGGGAGGCGGTCACTGGCTTGAGGAGGMAC026083_E41/42TCATGTCCCAGGTGACTAACACCACACAAGAAGGCATCTACTTCATCCTCACGGACAMSQVTNTTQEGIYFILTDIPGTCCCTGGATTTGAGGCCTCCCACATCTGGATCTCCATCCCCGTCTGCTGTCTCTACAFEASHIWISIPVCCLYTISIMCCATCTCCATCATGGGCAATACCACCATCCTCACTGTCATTCGCACAGAGCCATCTGGNTTILTVIRTEPSVHQRMYLTCCACCAGCGCATGTATCTGTTTCTCTCCATGCTGGCCCTGACGGACCTGGGTCTCAFLSMLALTDLGLTLTTLPTVMCCCTCACCACCCTACCCACAGTCATGCAGCTTCTCTGGTTCAACGTTCGTAGAATCAQLLWFNVRRISSEACFAQFFFGCTCTGAGGCCTGTTTTGCTCAGTTTTTCTTCCTTCATGGATTCTCCTTTATGGAGTLHGFSFMESSVLLAMSVDCYVCTTCTGTCCTCCTGGCTATGTCCGTTGACTGCTATGTGGCCATCTGCTGTCCCCTCCAICCPLHYASILTNEVIGRTGATTATGCCTCCATCCTCACCAATGAAGTCATTGGTAGAACTGGGTTAGCCATCATTTLAIICCCVLAVLPSLFLLKRLGCTGCTGTGTTCTGGCGGTTCTTCCCTCCCTTTTCTTACTCAAGCGACTGCCTTTCTPFCHSHLLSRSYCLHQDMIRLGCCACTCCCACCTTCTCTCTCGCTCCTATTGCCTCCACCAGGATATGATCCGCCTGGVCADIRLNSWYGFALALLIIITCTGTGCTGACATCAGGCTCAACAGCTGGTATGGATTTGCTCTTGCCTTGCTCATTAVDPLLIVISYTLILKNILGTATTATCGTGGATCCTCTGCTCATTGTGATCTCCTATACACTTATTCTGAAAAATATCTTWAERLRALNNCLSHILAVLVTGGGCACAGCCACCTGGGCTGAGCGACTCCGTGCCCTCAATAACTGCCTGTCCCACALYIPMVGVSMTHRFAKHASPLTTCTAGCTGTCCTGGTCCTCTACATTCCCATGGTTGGTGTATCTATGACTCATCGCTVHVIMANIYLLAPPVMNPIIYTTGCCAAGCATGCCTCTCCACTGGTCCATGTTATCATGGCCAATATCTACCTGCTGGSVKNKQIQWGMLNFLSLKNMHCACCCCCGGTGATGAACCCCATCATTTACAGTGTAAAGAACAAGCAGATCCAATGGGSRGAATGTTAAATTTCCTTTCCCTCAAAAATATGCATTCAAGATGAGGGAATGCATTTCTTAAATTACTGACAAGMAC026083_D43/44CTATCTATATTCTCCCACCATGCTGGGTCTCAATGGCACCCCCTTCCAGCCAGCAACMLGLNGTPFQPATLQLTGIPGACTCCAGCTGACAGGCATTCCTGGGATACAAACAGGCCTCACCTGGGTTGCCCTGATIQTGLTWVALIFCILYMISIVTTTCTGCATCCTCTACATGATCTCCATTGTAGGTAACCTCAGCATTCTCACTCTGGTGNLSILTLVFWEPALHQPMYYGTTTTGGGAGCCTGCTCTGCATCAGCCCATGTACTACTTCCTCTCTATGCTCGCTCTFLSMLALNDLGVSFSTLPTVICAATGATCTGGGAGTGTCCTTTTCTACACTTCCCACTGTGATTTCTACTTTCTGCTTSTFCFNYNHVAFNACLVQMFFCAACTACAACCATGTTGCGTTTAATGCTTGCCTGGTCCAGATGTTCTTCATCCACACIHTFSFMESGILLAMSLDRFVTTTCTCCTTCATGGAGTCAGGCATACTGCTGGCCATGAGCTTGGATCGCTTTGTGGCAICYPLRYVTVLTHNRILAMGTATTTGTTATCCATTACGCTATGTCACTGTGCTCACTCACAACCGTATATTGGCTATLGILTKSFTTLFPFPFVVKRLGGGTCTGGGCATCCTTACCAAGAGTTTCACCACTCTCTTCCCTTTCCCTTTTGTGGTPFCKGNVLHHSYCLHPDLMKVGAAACGACTGCCCTTCTGCAAAGGCAATGTTTTGCATCACTCCTACTGTCTCCATCCACGDIHVNNIYGLLVIIFTYGAGATCTCATGAAAGTAGCATGTGGAGACATCCATGTTAACAACATTTATGGGCTCTTMDSTFILLSYALILRAMLVIIGGTGATCATTTTTACCTATGGTATGGACTCAACTTTCATCCTGCTTTCCTACGCATTSQEQRLKALNTCMSHICAVLAGATCCTGAGAGCCATGCTGGTCATCATATCCCAGGAACAGCGGCTCAAGGCACTCAAFYVPIIAVSMIHRFWKSAPPVCACCTGCATGTCACACATCTGTGCAGTGCTGGCCTTTTATGTGCCCATAATTGCTGTVHVMMSNVYLFVPPMLNPIIYCTCCATGATTCACCGCTTCTGGAAAAGTGCTCCACCTGTTGTTCATGTCATGATGTCSVKTKEIRKGILKFFHKSQACAATGTCTACCTGTTTGTACCACCCATGCTCAACCCTATCATCTACAGTGTGAAAACCAAGGAGATCCGCAAAGGGATTCTCAAGTTCTTCCATAAATCCCAGGCCTGAGGAAGGCTGAAGGCCCGMAC026083_C45/46TTTATGCCATCTGCCTCTGCCATGATCATTTTCAACCTGAGCAGTTACAATCCAGGAMPSASAMIIFNLSSYNPGPFICCCTTCATTCTGGTAGGGATCCCAGGCCTGGAGCAATTCCATGTGTGGATTGGAATTLVGIPFLEQFHVWIGIPFCIICCCTTCTGTATCATCTACATTGTAGCTGTTGTGGGAAACTGCATCCTTCTCTACCTCYIVAVVGNCILLYLIVVEHSLATTGTGGTGGAGCATAGTCTTCATGAACCCATGTTCTTCTTTCTCTCCATGCTGGCCHEPMFFFLSMLAMTDLILSTAATGACTGACCTCATCTTGTCCACAGCTGGTGTGCCTAAAGCACTCAGTATCTTTTGGGVPKALSIFWLGAREITFPGCCTAGGGGCTCGCGAAATCACATTCCCAGGATGCCTTACACAAATGTTCTTCCTTCACLTQMFFLHYNFVLDSAILMAMTATAACTTTGTCCTGGATTCAGCCATTCTGATGGCCATGGCATTTGATCACTATGTAAFDHYVAICSPLRYTTILTPKGCTATCTGTTCTCCCTTGAGATATACCACCATCTTGACTCCCAAGACCATCATCAAGTIIKSAMGISFRSFCIILPDVAGTGCTATGGGCATCTCCTTTCGAAGCTTCTGCATCATCCTGCCAGATGTATTCTTGFLLTCLPFCRTRIIPHTYCEHCTGACATGCCTGCCTTTCTGCAGGACACGCATCATACCCCACACATACTGTGAGCATIGVAQLACADISINFWYGFCVATAGGTGTTGCCCAGCTCGCCTGTGCTGATATCTCCATCAACTTCTGGTATGGCTTTPIMTVISDVILIAVSYAHILCTGTGTTCCCATCATGACGGTCATCTCAGATGTGATTCTCATTGCTGTTTCCTACGCAAVFGLPSQDACQKALGTCGSHCACATCCTCTGTGCTGTCTTTGGCCTTCCCTCCCAAGATGCCTGCCAGAAAGCCCTCVCVILMFYTPAFFSILAHRFGGGCACTTGTGGTTCTCATGTCTGTGTCATCCTCATGTTTTATACACCTGCCTTTTTCHNVSRTFHIMFANLYIVIPPATCCATCCTCGCCCATCGCTTTGGACACAATGTCTCTCGCACCTTCCACATCATGTTTLNPMVYGVKTKQIRDKVILLFGCCAATCTCTACATTGTTATCCCACCTGCACTCAACCCCATGGTTTACGGAGTGAAGSKGTGACCAAGCAGATCAGAGATAAGGTTATACTTTTGTTTTCTAAGGGTACAGGATGATGMAC026083_B47/48GTCCTCAATTCATGCTGCTATCCAACATTACTCAGTTTAGCCCCATATTCTATCTCAMLLSNITQFSPIFYLTSFPGLCCAGCTTTCCTGGATTGGAAGGCATCAAACACTGGATTTTCATCCCCTTTTTCTTTAEGIKHWIFIPFFFMYMVAISGTGTACATGGTTGCCATCTCAGGCAATTGTTTCATTCTGATCATTATTAAGACCAACCNCFILIIIKTNPRLHTPMYYLCTCGTCTGCACACACCCATGTACTATCTACTATCCTTGCTGGCCCTCACTGACCTGGLSLLALTDLGLCVSTLPTTMGGGCTGTGTGTGTCCACGTTGCCCACCACTATGGGGATCTTCTGGTTTAACTCCCAGAIFWFNSQSIYFGACQIQMFCIGTATCTACTTTGGAGCGTGTCAAATCCAGATGTTCTGCATCCACTCTTTTTCCTTCAHSFSFMESSVLLMMSFDRFVATGGAGTCCTCAGTGCTCCTCATGATGTCCTTTGACCGCTTTGTGGCCATCTGCCACCICHPLRYSVIITGQQVVRAGLCTCTGAGGTATTCGGTCATTATCACTGGCCAGCAAGTGGTCAGAGCAGGCCTAATTGIVIFRGPVATIPIVLLLKAFPTCATCTTCCGGGGACCTGTGGCCACTATCCCTATTGTCCTCCTCCTGAAGGCTTTTCYCGSVVLSHSFCLHQEVIQLACCTACTGTGGATCTGTGGTCCTCTCCCACTCATTTTGCCTGCACCAGGAAGTGATACCTDTTFNNLYGLMVVVFTVMLAGCTGGCCTGCACAGATACCACCTTCAATAATCTGTATGGACTGATGGTGGTAGTTTDLVLIALSYGLILHTVAGLASTCACTGTGATGCTGGACCTGGTGCTCATCGCACTGTCCTATGGACTCATCCTGCACAQEEQRRAFQTCTAHLCAVLVFCAGTAGCAGGCCTGGCCTCCCAAGAGGAGCAGCGCCGTGCCTTTCAGACATGCACCGFVPMMGLSLVHRFGKHAPPAICTCATCTCTGTGCTGTGCTAGTATTCTTTGTGCCCATGATGGGGCTGTCCCTGGTGCHLLMANVYLFVPPMLNPIIYSACCGTTTTGGGAAGCATGCCCCACCTGCTATTCATCTTCTTATGGCCAATGTCTACCIKTEIHRAIIKLLGLKKASKTTTTTGTGCCTCCCATGCTTAACCCAATCATATACAGCATTAAGACCAAGGAGATCCACCGTGCCATTATCAAACTCCTAGGTCTTAAAAAGGCCAGTAAATGAGTCCTGGGGCTAAAACTCCCCCTAGAGGCCTATAAGAAGGCCCCAAATTGGACTGAAAATTTGGAGMAC026083_A49/50TTTGTTTTGCTATGGGGTTGTTCAATGTCACTCACCCTGCATTCTTCCTCCTGACTGMGLFNVTHPAFFLLTGIPGLEGTATCCCTGGTCTGGAGAGCTCTCACTCCTGGCTGTCAGGGCCCCTCTGCGTGATGTSSHSWLSGPLCVMYAVALGGNATGCTGTGGCCCTTGGGGGAAATACAGTGATCCTGCAGGCTGTGCGAGTGGAGCCCATVILQAVRVEPSLHEPMYYFLGCCTCCATGAGCCCATGTACTACTTCCTGTCCATGTTGTCCTTCAGTGATGTGGCCASMLSFSDVAISMATLPTVLRTTATCCATGGCCACACTGCCCACTGTACTCCGAACCTTCTGCCTCAATGCCCGCAACAFCLNARNITFDACLIQMFLIHTCACTTTTGATGCCTGTCTAATTCAGATGTTTCTTATTCACTTCTTCTCCATGATGGFFSMMESGILLAMSFDRYVAIAATCAGGTATTCTGCTGGCCATGAGTTTTGACCGCTATGTGGCCATTTGTGACCCCTCDPLRYATVLTTEVIAAMGLGTGCGCTATGCAACTGTGCTCACCACTGAAGTCATTGCTGCAATGGGTTTAGGTGCAGAAARSFITLFPLPFLIKRLPICTGCTCGAAGCTTCATCACCCTTTTCCCTCTTCCCTTTCTTATTAAGAGGCTGCCTACRSNVLSHSYCLHPDMMRLACTCTGCAGATCCAATGTTCTTTCTCACTCCTACTGCCTGCACCCAGACATGATGAGGCADISINSIYGLFVLVSTFGMDTTGCCTGTGCTGATATCAGTATCAACAGCATCTATGGACTCTTTGTTCTTGTATCCALFFIFLSYVLILRSVMATASRCCTTTGGCATGGACCTGTTTTTTATCTTCCTCTCCTATGTGCTCATTCTGCGTTCTGEERLKALNTCVSHILAVLAFYTCATGGCCACTGCTTCCCGTGAGGAACGCCTCAAAGCTCTCAACACATGTGTGTCACVPMIGVSTVHRFGKHVPCYIHATATCCTGGCTGTACTTGCATTTTATGTGCCAATGATTGGGGTCTCCACAGTGCACCVLMSNVYLFVPPVLNPLIYSAGCTTTGGGAAGCATGTCCCATGCTACATACATGTCCTCATGTCAAATGTGTACCTATKTKEIRRAIFRMFHHIKITTGTGCCTCCTGTGCTCAACCCTCTCATTTATAGCGCCAAGACAAAGGAAATCCGCCGAGCCATTTTCCGCATGTTTCACCACATCAAAATATGACTTTCACACTTGGCTTTAGAATCTCG56808-0151/52TTTGTTTTGCTATGGGGTTGTTCAATGTCACTCACCCTGCATTCTTCCTCCTGACTGMGLFNVTHPAFFLLTGIPGLEGTATCCCTGGTCTGGAGAGCTCTCACTCCTGGCTGTCAGGGCCCCTCTGCGTGATGTSSHSWLSGPLCVMYAVALGGNATGCTGTGGCCCTTGGGGGAAATACAGTGATCCTGCAGGCTGTGCGAGTGGAGCCCATVILQAVRVEPSLHEPMYYFLGCCTCCATGAGCCCATGTACTACTTCCTGTCCATGTTGTCCTTCAGTGATGTGGCCASMLSFSDVAISMATLPTVLRTTATCCATGGCCACACTGCCCACTGTACTCCGAACCTTCTGCCTCAATGCCCGCAACAFCLNARNITFDACLIQMFLIHTCACTTTTGATGCCTGTCTAATTCAGATGTTTCTTATTCACTTCTTCTCCATGATGGFFSMMESGILLAMSFDRYVAIAATCAGGTATTCTGCTGGCCATGAGTTTTGACCGCTATGTGGCCATTTGTGACCCCTCDPLRYATVLTTEVIAAMGLGTGCGCTATGCAACTGTGCTCACCACTGAAGTCATTGCTGCAATGGGTTTAGGTGCAGAAARSFITLFPLPFLIKRLPICTGCTCGAAGCTTCATCACCCTTTTCCCTCTTCCCTTTCTTATTAAGAGGCTGCCTACRSNVLSHSYCLHPDMMRLACTCTGCAGATCCAATGTTCTTTCTCACTCCTACTGCCTGCACCCAGACATGATGAGGCADISINSIYGLFVLVSTFGMDTTGCCTGTGCTGATATCAGTATCAACAGCATCTATGGACTCTTTGTTCTTGTATCCALFFIFLSYVLILRSVMATASRCCTTTGGCATGGACCTGTTTTTTATCTTCCTCTCCTATGTGCTCATTCTGCGTTCTGEERLKALNTCVSHILAVLAFYTCATGGCCACTGCTTCCCGTGAGGAACGCCTCAAAGCTCTCAACACATGTGTGTCACVPMIGVSTVHRFGKHVPCYIHATATCCTGGCTGTACTTGCATTTTATGTGCCAATGATTGGGGTCTCCACAGTGCACCVLMSNVYLFVPPVLNPLIYSAGCTTTGGGAAGCATGTCCCATGCTACATACATGTCCTCATGTCAAATGTGTACCTATKTKEIRRAIFRMFHHIKITTGTGCCTCCTGTGCTCAACCCTCTCATTTATAGCGCCAAGACAAAGGAAATCCGCCGAGCCATTTTCCGCATGTTTCACCACATCAAAATATGACTTTCACACTTGGCTTTAGAATCTGMAC027522_B53/54GACTAAATGATGGACAACCACTCTAGTGCCACTGAATTCCACCTTCTAGGCTTCCCTMMDNHSSATEFHLLGFPGSQGGGGTCCCAAGGACTACACCACATTCTTTTTGCTATATTCTTTTTCTTCTATTTAGTGLHHILFAIFFFFYLVTLMGNTACATTAATGGGAAACACGGTCATCATTGTGATTGTCTGTGTGGATAAACGTCTGCAGVIIVIVCVDKRLQSPMYFFLSTCCCCCATGTATTTCTTCCTCAGCCACCTCTCTACCCTGGAGATCCTGGTCACAACCHLSTLEILVTTIIVPMMLWGLATAATTGTCCCCATGATGCTTTGGGGATTGCTCTTCCTGGGATGCAGACAGTATCTTLFLGCRQYSLSHVSLNFSCGTTCTCTACATGTATCGCTCAACTTTTCCTGTGGGACCATGGAGTTTGCATTACTTGGAMEFALLGVMAVDRYVAVCNPLGTGATGGCTGTGGACCGTTATGTGGCTGTGTGTAACCCTTTGAGGTACAACATCATTRYNIIMNSSTCIWVVIVSWVFATGAACAGCAGTACCTGTATTTGGGTGGTAATAGTGTCATGGGTGTTTGGATTTCTTGFLSEIWPIYATFQFTFRKSNTCTGAAATCTGGCCCATCTATGCCACATTTCAGTTTACCTTCCGCAAATCAAATTCASLDHFYCDRGQLLKLSCDNTLTTAGACCATTTTTACTGTGACCGAGGGCAATTGCTCAAACTGTCCTGCGATAACACTLTEFILFLMAVFILIGSLIPTCTTCTCACAGAGTTTATCCTTTTCTTAATGGCTGTTTTTATTCTCATTGGTTCTTTGIVSYTYIISTILKIPSASGRRATCCCTACGATTGTCTCCTACACCTACATTATCTCCACCATCCTCAAGATCCCGTCAKAFSTFASHFTCVVIGYGSCLGCCTCTGGCCGGAGGAAAGCCTTCTCCACTTTTGCCTCCCACTTCACCTGTGTTGTGFLYVKPKQTQGVEYNKIVSLLATTGGCTATGGCAGCTGCTTGTTTCTCTACGTGAAACCCAAGCAAACACAGGGAGTTVSVLTPFLNPFIFTLRNDKVKGAGTACAATAAGATAGTTTCCCTGTTGGTTTCTGTGTTAACCCCCTTCCTGAATCCTVSVLTPFLNPFIFTLRNDKVKTTCATCTTTACTCTTCGGAATGACAAAGTCAAAGAGGCCCTCCGAGATGGGATGAAACGCTGCTGTCAACTCCTGAAAGATTAGCTCG56290-0155/56GACTAAATGATGGACAACCACTCTAGTGCCACTGAATTCCACCTTCTAGGCTTCCCTMMDNHSSATEFHLLGFPGSQGGGGTCCCAAGGACTACACCACATTCTTTTTGCTATATTCTTTTTCCTCTATTTAGTGLHHILFAIFFFFYLVTLMGNTACATTAATGGGAAACACGGTCATCATTGTGATTGTCTGTGTGGATAAACGTCTGCAGVIIVIVCVDKRLQSPMYFFLSTCCCCCATGTATTTCTTCCTCAGCCACCTCTCTACCCTGGAGATCCTGGTCACAACCHLSTLEILVTTIIVPMMLWGLATAATTGTCCCCATGATGCTTTGGGGATTGCTCTTCCTGGGATGCAGACAGTATCTTLFLGCRQYLSLHVSLNFSCGTTCTCTACATGTATCGCTCAACTTTTCCTGTGGGACCATGGAGTTTGCATTACTTGGAMEFALLGVMAVDRYVAVCNPLGTGATGGCTGTGGACCGTTATGTGGCTGTGTGTAACCCTTTGAGGTACAACATCATTRYNIIMNSSTCIWVVIVSWVFATGAACAGCAGTACCTGTATTTGGGTGGTAATAGTGTCATGGGTGTTTGGATTTCTTGFLSEIWPIYATFQFTFRKSNTCTGAAATCTGGCCCATCTATGCCACATTTCAGTTTACCTTCCGCAAATCAAATTCASLDHFYCDRGQLLKLSCDNTLTTAGACCATTTTTACTGTGACCGAGGGCAATTGCTCAAACTGTCCTGCGATAACACTLTEFILFLMAVFILIGSLIPTCTTCTCACAGAGTTTATCCTTTTCTTAATGGCTGTTTTTATTCTCATTGGTTCTTTGIVSYTYIISTILKIPSASGRRATCCCTACGATTGTCTCCTACACCTACATTATCTCCACCATCCTCAAGATCCCGTCAKAFSTFASHFTCVVIGYGSCLGCCTCTGGCCGGAGGAAAGCCTTCTCCACTTTTGCCTCCCACTTCACCTGTGTTGTGFLYVKPKQTQGVEYNKIVSLLATTGGCTATGGCAGCTGCTTGTTTCTCTACGTGAAACCCAAGCAAACACAGGGAGTTVSVLTPFLNPFIFTLRNDKVKGAGTACAATAAGATAGTTTCCCTGTTGGTTTCTGTGTTAACCCCCTTCCTGAATCCTEALRDGMKRCCQLLKDTTCATCTTTACTCTTCGGAATGACAAAGTCAAAGAGGCCCTCCGAGATGGGATGAAACGCTGCTGTCAACTCCTGAAAGATTAGCTGMAC036216_D57/58GACTAAATGATGGACAACCACTCTAGTGCCACTGAATTCCACCTTCTAGGCTTCCCTMMDNHSSATEFHLLGFPGSQGGGGTCCCAAGGACTACACCACATTCTTTTTGCTATATTCTTTTTCTTCTATTTAGTGLHHILFAIFFFFYLVTLMGNTACATTAATGGGAAACACGGTCATCATTGTGATTGTCTGTGTGGATAAACGTCTGCAGVIIVIVCVDKRLQSPMYFFLSTCCCCCATGTATTTCTTCCTCAGCCACCTCTCTACCCTGGAGATCCTGGTCACAACCHLSTLEILVTTIIVPMMLWGLATAATTGTCCCCATGATGCTTTGGGGATTGCTCTTCCTGGGATGCAGACAGTATCTTLFLGCRQYLSLHVSLNFSCGTTCTCTACATGTATCGCTCAACTTTTCCTGTGGGACCATGGAGTTTGCATTACTTGGAMEFALLGVMAVDRYVAVCNPLGTGATGGCTGTGGACCGTTATGTGGCTGTGTGTAACCCTTTGAGGTACAACATCATTRYNIIMNSSTCIWVVIVSWVFATGAACAGCAGTACCTGTATTTGGGTGGTAATAGTGTCATGGGTGTTTGGATTTCTTGFLSEIWPIYATFQFTFRKSNTCTGAAATCTGGCCCATCTATGCCACATTTCAGTTTACCTTCCGCAAATCAAATTCASLDHFYCDRGQLLKLSCDNTLTTAGACCATTTTTACTGTGACCGAGGGCAATTGCTCAAACTGTCCTGCGATAACACTLTEFILFLMAVFILIGSLIPTCTTCTCACAGAGTTTATCCTTTTCTTAATGGCTGTTTTTATTCTCATTGGTTCTTTGIVSYTYIISTILKIPSASGRRATCCCTACGATTGTCTCCTACACCTACATTATCTCCACCATCCTCAAGATCCCGTCAKAFSTFASHFTCVVIGYGSCLGCCTCTGGCCGGAGGAAAGCCTTCTCCACTTTTGCCTCCCACTTCACCTGTGTTGTGFLYVKPKQTQGVEYNKIVSLLATTGGCTATGGCAGCTGCTTGTTTCTCTACGTGAAACCCAAGCAAACACAGGGAGTTVSVLTPFLNPFIFTLRNDKVKGAGTACAATAAGATAGTTTCCCTGTTGGTTTCTGTGTTAACCCCCTTCCTGAATCCTEALRDGMKRCCQLLKDTTCATCTTTACTCTTCGGAATGACAAAGTCAAAGAGGCCCTCCGAGATGGGATGAAACGCTGCTGTCAACTCCTGAAAGATTAGCTGMAC073079_A59/60GACTAAATGATGGACAACCACTCTAGTGCCACTGAATTCCACCTTCTAGGCTTCCCTMMDNHSSATEFHLLGFPGSQGGGGTCCCAAGGACTACACCACATTCTTTTTGCTATATTCTTTTTCTTCTATTTAGTGLHHILFAIFFFFYLVTLMGNTACATTAATGGGAAACACGGTCATCATTGTGATTGTCTGTGTGGATAAACGTCTGCAGVIIVIVCVDKRLQSPMYFFLSTCCCCCATGTATTTCTTCCTCAGCCACCTCTCTACCCTGGAGATCCTGGTCACAACCHLSTLEILVTTIIVPMMLWGLATAATTGTCCCCATGATGCTTTGGGGATTGCTCTTCCTGGGATGCAGACAGTATCTTLFLGCRQYLSLHVSLNFSCGTTCTCTACATGTATCGCTCAACTTTTCCTGTGGGACCATGGAGTTTGCATTACTTGGAMEFALLGVMAVDRYVAVCNPLGTGATGGCTGTGGACCGTTATGTGGCTGTGTGTAACCCTTTGAGGTACAACATCATTRYNIIMNSSTCIWVVIVSWVFATGAACAGCAGTACCTGTATTTGGGTGGTAATAGTGTCATGGGTGTTTGGATTTCTTGFLSEIWPIYATFQFTFRKSNTCTGAAATCTGGCCCATCTATGCCACATTTCAGTTTACCTTCCGCAAATCAAATTCASLDHFYCDRGQLLKLSCDNTLTTAGACCATTTTTACTGTGACCGAGGGCAATTGCTCAAACTGTCCTGCGATAACACTLTEFILFLMAVFILIGSLIPTCTTCTCACAGAGTTTATCCTTTTCTTAATGGCTGTTTTTATTCTCATTGGTTCTTTGIVSYTYIISTILKIPSASGRRATCCCTACGATTGTCTCCTACACCTACATTATCTCCACCATCCTCAAGATCCCGTCAKAFSTFASHFTCVVIGYGSCLGCCTCTGGCCGGAGGAAAGCCTTCTCCACTTTTGCCTCCCACTTCACCTGTGTTGTGFLYVKPKQTQGVEYNKIVSLLATTGGCTATGGCAGCTGCTTGTTTCTCTACGTGAAACCCAAGCAAACACAGGGAGTTVSVLTPFLNPFIFTLRNDKVKGAGTACAATAAGATAGTTTCCCTGTTGGTTTCTGTGTTAACCCCCTTCCTGAATCCTEALRDGMKRCCQLLKDTTCATCTTTACTCTTCGGAATGACAAAGTCAAAGAGGCCCTCCGAGATGGGATGAAACGCTGCTGTCAACTCCTGAAAGATTAGCTGMAP002418_A61/62CCATGCAGAGGAGCAATCATACAGTGACTGAGTTTATACTGCTGGGCTTCACCACAGMQRSNHTVTEFILLGFTTDPGACCCAGGAATGCAGCTGGGCCTCTTCGTGGTGTTCCTGGGCGTGTACTCTCTCACTGMQLGLFVVFLGVYSLTVVGNSTGGTAGGAAATAGCACCCTCATCGTGTTGATCTGTAATGACTCCTGCCTCCACACACTLIVLICNDSCLHTPMYFVVGCCATGTATTTTGTCGTTGGAAATCTGTCGTTTCTGGATCTCTGGTATTCTTCTGTCTNLSFLDLWYSSVYTPKILVTCACACCCCAAAGATCCTAGTGACCTGCATCTCTGAAGACAAAAGCATCTCCTTTGCTGISEDKSISFAGCLCQFFFSAGGCTGCCTGTGTCAGTTCTTCTTCTCTGCAGGGCTGGCCTATAGTGAGTGCTACCTGCLAYSECYLLAAVAYDRYVAISTGGCTGCCGTGGCTTATGACCGCTACGTGGCCATCTCCAAGCCCCTGCTTTATGCTCKPLLYAQAMSIKLCALLVAVSAGGCCATGTCCATAAAGCTGTGTGCATTGCTGGTAGCAGTCTCATATTGTGGTGGCTYCGGFINSSIITKKTFSFNFCTTATTAACTCTTCAATCATCACCAAGAAAACGTTTTCCTTTAACTTCTGCCGTGAAARENIIDDFFCDLLPLVELACGACATCATTGATGACTTTTTCTGTGATTTGCTTCCCTTGGTGGAGCTGGCCTGTGGCGEKGGYKIMMYFLLASNVICPAAGAAGGGCGGCTATAAAATTATGATGTACTTCCTGCTGGCCTCCAATGTCATCTGCCVLILASYLFIITSVLRISSSKCCGCAGTGCTCATCCTGGCCTCCTACCTCTTTATCATCACCAGTGTCTTGAGGATCTGYLKAFSTCSSHLTSVTLYYGCCTCCTCCAAGGGCTACCTCAAAGCCTTCTCCACATGCTCCTCCCACCTGACCTCTGSILYIYALPRSSYSFDMDKIVTCACTTTATACTATGGCTCCATTCTCTACATCTACGCTCTCCCCAGATCTAGCTATTSTFYTVVFPMLNLMIYSLRNKCTTTTGATATGGACAAAATAGTTTCTACATTTTACACTGTGGTATTCCCCATGTTGADVKEALKKLLPATCTCATGATCTACAGCCTAAGGAATAAGGATGTGAAAGAGGCTCTGAAAAAACTTCTCCCATAAATCAAGATTATCTCCACCAGAGGAGAAACAAAGACTATTTTAGATGCAGTTTTTTGMAP002345_B63/64TGATGGAAAATAAGACAGAAGTAACACAATTCATTCTTCTAGGACTAACCAATGACTMENKTEVTQFILLGLTNDSELCAGAACTGCAGGTTCCCCTCTTTATAACGTTCCCCTTCATCTATATTATCACTCTGGQVPLFITFPFIYIITLVGNLGTTGGAAACCTGGGAATTATTGTATTGATATTCTGGGATTCCTGTCTCCACAATCCCAIIVLIFWDSCLHNPMYFFLSNTGTACTTTTTTCTCAGTAACTTGTCTCTAGTGGACTTTTGCTACTCTTCAGCTGTCALSLVDFCYSSAVYPIVMAGFLCTCCCATCGTCATGGCTGGATTCCTTATAGAAGACAAGGTCATCTCTTACAATGCATIEDKVISYNACAAQMYIFVAFGTGCTGCTCAAATGTATATCTTTGTAGCTTTTGCCACTGTGGAAAATTACCTCTTGGATVENYLLASMAYDRYAAVCKCCTCAATGGCCTATGACCGCTATGCAGCAGTGTGCAAACCCCTACATTACACCACAAPLHYTTTMTTTVCARLAIGSYCCATGACAACAACTGTGTGTGCTCGTCTGGCCATAGGCTCCTACCTCTGTGGTTTCCLCGFLNASIHTGDTFSLSFCKTGAATGCCTCCATCCACACTGGGGACACATTTAGTCTCTCTTTCTGTAAGTCCAATGSNEVHHFFCDIPAVMVLSCSDAAGTCCATCACTTTTTCTGTGATATTCCAGCAGTCATGGTTCTCTCTTGCTCTGATARHISELVLIYVVSFNIFIALLGACATATTAGCGAGCTTGTTCTTATTTATGTTGTGAGCTTCAATATCTTTATAGCTCVILISYTFIFITILKMHSASVTCCTGGTTATCTTGATATCCTACACATTCATTTTTATCACCATCCTAAAGATGCACTYQKPLSTCASHFIAVGIFYGTCAGCTTCAGTATACCAGAAGCCTTTGTCCACCTGTGCCTCTCATTTCATTGCAGTCGIIFMYLQPSSSHSMDTDKMAPGCATCTTCTATGGGACTATTATCTTCATGTACTTACAACCCAGCTCCAGTCACTCCAVFYTMVIPMLNPLVYSLRNKETGGACACAGACAAAATGGCACCTGTGTTCTATACAATGGTCATCCCCATGCTGAACCVFYTMVIPMLNPLVYSLRNKECTCTGGTCTATAGTCTGAGGAACAAGGAAGTGAAGAGTGCATTCAAGAAAGTTGTTGAGAAGGCAAAATTGTCTGTAGGATGGTCAGTTTAACATTGMAL391156_A65/66ACAATGGATGTGGGCAATAAGTCTACCATGTCTGAATTTGTTTTGCTGGGGCTCTCTMDVGNKSTMSEFVLLGLSNSWAATTCCTGGGAACTACAGATGTTTTTCTTTATGGTGTTTTCATTGCTTTATGTGGCAELQMFFFMVFSLLYVATMVGNACAATGGTGGGTAACAGCCTCATAGTCATCACAGTTATAGTGGACCCTCACCTACACSLIVITVIVDPHLHSPMYFLLTCTCCTATGTATTTCCTGCTTACCAATCTTTCAATCATTGATATGTCTCTTGCTTCTTNLSIIDMSLASFATPKMITDTTCGCCACCCCAAAGATGATTACAGATTACCTAACAGGTCACAAAACCATCTCTTTTYLTGHKTISFDGCLTQIFFLGGATGGCTGCCTTACCCAGATATTCTTTCTCCACCTTTTCACTGGAACTGAGATCATCLFTGTIEELLMAMSFDRYIAITTACTCATGGCCATGTCCTTTGATAGGTATATTGCAATATGCAAGCCCCTGCACTATCKPLHYASVISPQVCVALVVAGCTTCTGTCATTAGTCCCCAGGTGTGTGTTGCTCTCGTGGTGGCTTCCTGGATTATGSWIMGVMHSMSQVIFALTLPFGGAGTTATGCATTCAATGAGTCAGGTCATATTTGCCCTCACGTTACCATTCTGTGGTCGPYEVDSFFCDLPVVFQLACCCCTATGAGGTAGACAGCTTTTTCTGTGACCTTCCTGTGGTGTTCCAGTTGGCTTGTVDTYVLGLFMISTSGIIALSCGTGGATACTTATGTTCTGGGCCTCTTTATGATCTCAACAAGTGGCATAATTGCGTTGFIVLFNSYVIVLVTVKHHSSRTCCTGTTTTATTGTTTTATTTAATTCATATGTTATTGTCCTGGTTACTGTGAAGCATGSSKALSTCTAHFIVVFLFFGCATTCTTCCAGAGGATCATCTAAGGCCCTTTCTACTTGTACAGCTCATTTCATTGTTPCIFIYMWPLSSFLTDKILSVGTCTTCTTGTTCTTTGGGCCATGCATCTTCATCTACATGTGGCCACTAAGCAGCTTTFYTIFTPTLNPIIYTLRNQEVCTCACAGACAAGATTCTGTCTGTGTTTTATACCATCTTTACTCCCACTCTGAACCCAKIAMRKLKNRFLNFNKAMPSATAATCTATACTTTGAGGAATCAAGAAGTAAAGATAGCCATGAGGAAACTGAAAAATAGGTTTCTAAATTTTAATAAGGCAATGCCTTCATAGTTTTTCG55962-0167/68ACAATGGATGTGGGCAATAAGTCTACCATGTCTGAATTTGTTTTGCTGGGGCTCTCTMDVGNKSTMSEFVLLGLSNSWAATTCCTGGGAACTACAGATGTTTTTCTTTATGGTGTTTTCATTGCTTTATGTGGCAELQMFFFMVFSLLYVATMVGNACAATGGTGGGTAACAGCCTCATAGTCATCACAGTTATAGTGGACCCTCACCTACACSLIVITVIVDPHLHSPMYFLLTCTCCTATGTATTTCCTGCTTACCAATCTTTCAATCATTGATATGTCTCTTGCTTCTTNLSIIDMSLASFATPKMITDTTCGCCACCCCAAAGATGATTACAGATTACCTAACAGGTCACAAAACCATCTCTTTTYLTGHKTISFDGCLTQIFFLHGATGGCTGCCTTACCCAGATATTCTTTCTCCACCTTTTCACTGGAACTGAGATCATCLFTGTEIILLMAMSFDRYIAITTACTCATGGCCATGTCCTTTGATAGGTATATTGCAATATGCAAGCCCCTGCACTATCKPLHYASVISPQVCVALVVAGCTTCTGTCATTAGTCCCCAGGTGTGTGTTGCTCTCGTGGTGGCTTCCTGGATTATGSWIMGVMHSMSQVIFALTLPFGGAGTTATGCATTCAATGAGTCAGGTCATATTTGCCCTCACGTTACCATTCTGTGGTCGPYEVDSFFCDLPVVFQLACCCCTATGAGGTAGACAGCTTTTTCTGTGACCTTCCTGTGGTGTTCCAGTTGGCTTGTVDTYVLGLFMISTSGIIALSCGTGGATACTTATGTTCTGGGCCTCTTTATGATCTCAACAAGTGGCATAATTGCGTTGFIVLFNSYVIVLVTVKHHSSRTCCTGTTTTATTGTTTTATTTAATTCATATGTTATTGTCCTGGTTACTGTGAAGCATGSSKALSTCTAHFIVVFLFFGCATTCTTCCAGAGGATCATCTAAGGCCCTTTCTACTTGTACAGCTCATTTCATTGTTPCIFIYMWPLSSFLTDKILSVGTCTTCTTGTTCTTTGGGCCATGCATCTTCATCTACATGTGGCCACTAAGCAGCTTTFYTIFTPTLNPIIYTLRNQEVCTCACAGACAAGATTCTGTCTGTGTTTTATACCATCTTTACTCCCACTCTGAACCCAKIAMRKLKNRFLNFNKAMPSATAATCTATACTTTGAGGAATCAAGAAGTAAAGATAGCCATGAGGAAACTGAAAAATAGGTTTCTAAATTTTAATAAGGCAATGCCTTCATAGTTTTTGMAC019093_A69/70GATGATTGAATGGAGATGGAAAACTGCACCAGGGTAAAAGAATTTATTTTCCTTGGCMEMENTCRVKEFIFLGLTQNRCTGACCCAGAATCGGGAAGTGAGCTTAGTCTTATTTCTTTTCCTACTCTTGGTGTATEVSLVLFLFLLLVYVTTLLGNGTGACAACTTTGCTGGGAAACCTCCTCATCATGGTCACTGTTACCTGTGAATCTCGCLLIMVTVTCESRLHTPMYFLLCTTCACACGCCCATGTATTTTTTGCTCCATAATTTATCTATTGCCGATATCTGCTTCHNLSIAKICFSSITVPKVLVDTCTTCCATCACAGTGCCCAAGGTTCTGGTGGACCTTCTGTCTGAAAGAAAGACCATCLLSERKTISFNHCFTQMFLFHTCCTTCAATCATTGCTTCACTCAGATGTTTCTATTCCACCTTATTGGAGGGGTGGATLIGGVDVFSLSVMALDRYVAIGTATTTTCTCTTTCGGTGATGGCATTGGATCGATATGTGGCCATCTCCAAGCCCCTGSKPLHYATIMSRDHCIGLTVACACTATGCGACTATCATGAGTAGAGACCATTGCATTGGGCTCACAGTGGCTGCCTGGAWLGGFVHSIVQISLLLPLPFTTGGGGGGCTTTGTCCACTCCATCGTGCAGATTTCCCTGTTGCTCCCACTCCCTTTCCGPNVLDTFYCDVHRVLKLAHTGCGGACCCAATGTTCTTGACACTTTCTACTGTGATGTCCACCGGGTCCTCAAACTGTDIFILELLMISNNGLLTTLWGCCCATACAGACATTTTCATACTTGAACTACTAATGATTTCCAACAATGGACTGCTCFFLLLVSYIVILSLPKLQAGEACCACACTGTGGTTTTTCCTGCTCCTGGTGTCCTACATAGTCATATTATCATTACCCGRRKAISTCTSHITVVTLHFVAAGTCTCAGGCAGGAGAGGGCAGGAGGAAAGCCATCTCCACCTGCACCTCCCACATCPCIYVYARPFTALPMDKAISVACTGTGGTGACCCTGCATTTCGTGCCCTGCATCTATGTCTATGCCCGGCCCTTCACTTFTVISPLLNPLIYTLRNHEMGCCCTCCCCATGGATAAGGCCATCTCTGTCACCTTCACTGTCATCTCCCCTCTGCTCKSAMRRLKRRLVPSDRKAACCCCTTGATCTACACTCTGAGGAACCATGAGATGAAGTCAGCCATGAGGAGACTGAAGAGAAGACTTGTGCCTTCTGATAGAAAATAGAAAAAAAAATCCTCAGCTCTGMAC022207_A71/72ATATGATCTGTGAAAATCACACCAGAGTCACTGAATTTATTCTTCTTGGTTTTACAAMICENHTRVTEFILLGFTNNPACAACCCCGAGATGCAAGTTTCCCTCTTTATTTTTTTCCTGGCCATTTATACAGTCAEMQVSLFIFFLAIYTVTLLGNCTTTGTTGGGCAACTTTCTTATTGTCACAGTTACCAGTGTGGATCTCGCACTTCAAAFLIVTVTSVDLALQTPMYFFLCACCCATGTACTTCTTTCTTCAAAATCTGTCACTTCTTGAAGTATGTTTCACCTTGGQNLSLLEVCFTLVMVPKMLVDTTATGGTGCCAAAAATGCTTGTAGATCTAGTGTCCCCAAGGAAAATTATCTCTTTTGLVSPRKIISFVGCGTQMYFFFTGGGCTGTGGTACCCAGATGTACTTCTTCTTCTTCTTTGGCAGTTCTGAATGTTTCCFFGSSECFLLSMMAYDRFVAITTCTCTCCATGATGGCTTATGATCGCTTTGTGGCCATCTGTAACCCTCTCCATTATTCNPLHYSVIMNRSLCLWMAIGCAGTCATAATGAACAGGTCCCTATGCTTGTGGATGGCCATAGGCTCTTGGATGTCCGSWMSGVPVSMLQTAWMMALPFGTGTTCCTGTGTCTATGCTACAGACAGCTTGGATGATGGCCCTTCCTTTCTGTGGACCGPNAVDHFFCDGPPVLKLVTCAAATGCCGTGGACCACTTTTTCTGTGATGGTCCCCCAGTGTTAAAACTAGTCACAGVDTTMYEMQALASTLLFIMFPTGGATACAACCATGTATGAAATGCAAGCACTTGCCTCCACACTCCTGTTTATCATGTFCLILVSYTRIIITILRMSSATTCCCTTTTGTCTCATTTTGGTTTCCTACACCCGCATTATCATAACAATTCTGAGGATGRQKAFSTCSSHLIVVSLFYTGTCCTCTGCCACTGGCCGCCAGAAGGCATTTTCTACTTGTTCCTCACACCTCATTGGTASLTYLRPKSNQSPESKKLTGGTGTCCCTCTTCTACGGAACAGCCAGTCTGACCTACCTGCGGCCCAAATCAAACCVLSLYTVITPMLNPIIYGLRNAGTCCCCTGAGAGCAAGAAGCTAGTGTCATTGTCCTACACTGTCATCACACCTATGCNEVKGAVKRTITQKVLQKLDVTAAACCCCATCATCTACGGCCTGAGGAACAATGAAGTGAAAGGGGCTGTCAAGAGGAFCAATCACTCAAAAAGTCTTACAGAAGTTAGATGTGTTTTGACTTCTATTCG55769-0273/74ATATGATCTGTGAAAATCACACCAGAGTCACTGAATTTATTCTTCTTGGTTTTACAAMICENHTRVTEFILLGFTNNPACAACCCCGAGATGCAAGTTTCCCTCTTTATTTTTTTCCTGGCCATTTATACAGTCAEMQVSLFIFFLAIYTVTLLGNCTTTGTTGGGCAACTTTCTTATTGTCACAGTTACCAGTGTGGATCTCGCACTTCAAAFLIVTVTSVDLALQTPMYFFLCACCCATGTACTTCTTTCTTCAAAATCTGTCACTTCTTGAAGTATGTTTCACCTTGGQNLSLLEVCFTLVMVPKMLVDTTATGGTGCCAAAAATGCTTGTAGATCTAGTGTCCCCAAGGAAAATTATCTCTTTTGLVSPRKIISFVGCGTQMYFFFTGGGCTGTGGTACCCAGATGTACTTCTTCTTCTTCTTTGGCAGTTCTGAATGTTTCCFFGSSECFLLSMMAYDRFVAITTCTCTCCATGATGGCTTATGATCGCTTTGTGGCCATCTGTAACCCTCTCCATTATTCNPLHYSVIMNRSLCLWMAIGCAGTCATAATGAACAGGTCCCTATGCTTGTGGATGGCCATAGGCTCTTGGATGTCCGSWMSGVPVSMLQTAWMMALPFGTGTTCCTGTGTCTATGCTACAGACAGCTTGGATGATGGCCCTTCCTTTCTGTGGACCGPNAVDHFFCDGPPVLKLVTCAAATGCCGTGGACCACTTTTTCTGTGATGGTCCCCCAGTGTTAAAACTAGTCACAGVDTTMYEMQALASTLLFIMFPTGGATACAACCATGTATGAAATGCAAGCACTTGCCTCCACACTCCTGTTTATCATGTFCLILVSYTRIIITILRMSSATTCCCTTTTGTCTCATTTTGGTTTCCTACACCCGCATTATCATAACAATTCTGAGGATGRQKAFSTCSSHLIVVSLFYTGTCCTCTGCCACTGGCCGCCAGAAGGCATTTTCTACTTGTTCCTCACACCTCATTGGTASLTYLRPKSNQSPESKKLTGGTGTCCCTCTTCTACGGAACAGCCAGTCTGACCTACCTGCGGCCCAAATCAAACCVLSLYTVITPMLNPIIYGLRNAGTCCCCTGAGAGCAAGAAGCTAGTGTCATTGTCCTACACTGTCATCACACCTATGCNEVKGAVKRTITQKVLQKLDVTAAACCCCATCATCTACGGCCTGAGGAACAATGAAGTGAAAGGGGCTGTCAAGAGGAFCAATCACTCAAAAAGTCTTACAGAAGTTAGATGTGTTTTGACTTCTATTGMAC010760_A75/76AAATGAAGATAGCAAACAACACAGTAGTGACAGAATTTATCCTCCTTGGTCTGACTCMKIANNTVVTEFILLGLTQSQAGTCTCAAGATATTCAGCTCTTGGTCTTTGTGCTGATCTTAATTTTCTACCTTATCADIQLLVFVLILIFYLIILPGNTCCTCCCTGGAAATTTTCTCATTATTTTCACCATAAGGTCAGACCCTGGGCTCACAGFLIIFTIRSDPGLTAPLYLFLCCCCCCTCTATTTATTTCTGGGCAACTTGGCCTTCCTGGATGCATCCTACTCCTTCAGNLAFLDASYSFIVAPRMLVDTTGTGGCTCCCAGGATGTTGGTGGACTTCCTCTCTGAGAAAAAGGTAATCTCCTACAFLSEKKVISYRGCITQLFFLHGAGGCTGCATCACTCAGCTCTTTTTCTTGCACTTCCTTGGAGGAGGGGAGGGATTACFLGGGEGLLLVVMAFDRYIAITCCTTGTTGTGATGGCCTTTGACCGCTACATCGCCATCTGCCGGCCTCTGCACTGTTCRPLHCSTVMNPRACYAMMLACAACTGTCATGAACCCTAGAGCCTGCTATGCAATGATGTTGGCTCTGTGGCTTGGGGLWLGGFVHSIIQVVLILRLPFGTTTTGTCCACTCCATTATCCAGGTGGTCCTCATCCTCCGCTTGCCTTTTTGTGGCCCGPNQLDNFFCDVRQVIKLACCAAACCAGCTGGACAACTTCTTCTGTGATGTCCGACAGGTCATCAAGCTGGCTTGCATDMFVVELLMVFNSGLMTLLCCCGACATGTTTGTGGTGGAGCTTCTAATGGTCTTCAACAGTGGCCTGATGACACTCCFLGLLASYAVILCHVRRAASETGTGCTTTCTGGGGCTTCTGGCTTCCTATGCAGTCATCCTCTGCCATGTTCGTAGGGGKNKAMSTCTTRVIIILLMFGCAGCTTCTGAAGGGAAGAACAAGGCCATGTCCACGTGCACCACTCGTGTCATTATTAPAIFIYMCPFRALPADKMVSLTACTTCTTATGTTTGGACCTGCTATCTTCATCTACATGTGCCCTTTCAGGGCCTTACFHTVIFPLMNPMIYTLRNQEVCAGCTGACAAGATGGTTTCTCTCTTTCACACAGTGATCTTTCCATTGATGAATCCTAKTSMKRLLSRHVVCQVDFIIRTGATTTATACCCTTCGCAACCAGGAAGTGAAAACTTCCATGAAGAGGTTATTGAGTCNGACATGTAGTCTGTCAAGTGGATTTTATAATAAGAAACTGAGAAGGAGGAATTCTGGCTGGAATTCATATCATTCATTTAACAAGTCCTGTTTTTCACTGACG55993-0177/78ACTGCCTCAATTTACTTCAGGATTTTGGAGGGCACCCACCTTCCCCCTTGTCTCCTCMTLGSLGNSSSSVSATFLLSGACACAATGACCCTGGGATCCCTGGGAAACAGCAGCAGCAGCGTTTCTGCTACCTTCCIPGLERMHIWISIPLCFMYLVTGCTGAGTGGCATCCCTGGGCTGGAGCGCATGCACATCTGGATCTCCATCCCACTGTSIPGNCTILFIIKTERSLHEPGCTTCATGTATCTGGTTTCCATCCCGGGCAACTGCACAATTCTTTTTATCATTAAAAMYLFLSMLALIDLGLSLCTLPCAGAGCGCTCACTTCATGAACCTATGTATCTCTTCCTGTCCATGCTGGCTCTGATTGTVLGIFWVGAREISHDACFAQACCTGGGTCTCTCCCTTTGCACTCTCCCTACAGTCCTGGGCATCTTTTGGGTTGGAGLFFIHCFSFLESSVLLSMAFDCACGAGAAATTAGCCATGATGCCTGCTTTGCTCAGCTCTTTTTCATTCACTGCTTCTRFVAICHPLHYVSILTNTVIGCCTTCCTCGAGTCCTCTGTGCTACTGTCTATGGCCTTTGACCGCTTTGTGGCTATCTRIGLVSLGRSVALIFPLPFMLGCCACCCCTTGCACTATGTTTCCATTCTCACCAACACAGTCATTGGCAGGATTGGCCKRFPYCGSPVLSHSYCLHQEVTGGTCTCTCTGGGTCGTAGTGTAGCACTCATTTTTCCATTACCTTTTATGCTCAAAAVKLACADMKANSIYGMFVIVSGATTCCCCTATTGTGGCTCCCCAGTTCTCTCACATTCTTATTGTCTCCACCAAGAAGTVGIDSLLILFSYALILRTVLTGGTGAAATTGGCCTGTGCCGACATGAAGGCCAACAGCATCTACGGCATGTTTGTCASIASRAERFKALNTCVSHICATCGTCTCTACAGTGGGTATAGACTCACTGCTCATCCTCTTCTCTTATGCTCTGATCCVLLFYTPMIGLSVIHRFGKQATGCGCACCGTGCTGTCCATCGCCTCCAGGGCTGAGAGATTCAAGGCCCTTAACACCTPHLVQVVMGFMYLLFPPVMNPGTGTTTCCCACATCTGTGCTGTGCTGCTCTTCTACACTCCCATGATTGGCCTCTCTGIVYSVKTKQIRDRVTHAFCYTCATCCATCGCTTTGGAAAGCAGGCACCTCACCTGGTCCAGGTGGTCATGGGTTTCATGTATCTTCTCTTTCCTCCTGTGATGAATCCCATTGTCTACAGTGTGAAGACCAAACAGATCCGGGATCGAGTGACGCATGCCTTTTGTTACTAACCG56038-0179/80ATAATACTGATGGAGAATTGTACGGAAGTGACAAAGTTCATTCTTCTAGGACTAACCMENCTEVTKFILLGLTSVPELAGTGTCCCAGAACTACAGATCCCCCTCTTTATCTTGTTCACCTTCATCTACCTCCTCQIPLFILFTFIYLLTLCGNLGACTCTGTGTGGGAACCTGGGGATGATGTTGCTGATCCTGATGGACTCTTGTCTCCACMMLLILMDSCLHTPMYFFLSNACCCCCATGTACTTTTTCCTCAGTAACCTGTCTCTGGTGGACTTTGGATACTCCTCALSLVDFGYSSAVTPKVMAGFLGCTGTCACTCCCAAGGTCATGGCTGGGTTCCTTAGAGGAGACAAGGTCATCTCCTACRGDKVISYNACAVQMFFFVALAATGCATGTGCTGTTCAGATGTTCTTCTTTGTAGCCTTGGCCACGGTGGAAAATTACATVENYLLASMAYDRYAAVCKTTGTTGGCCTCAATGGCCTATGACCGCTATGCAGCAGTGTGCAAACCCCTACACTACPLHYTTTMTASVGACLALGSYACCACCACCATGACGGCCAGTGTAGGTGCCTGTCTGGCCCTAGGCTCATATGTCTGTVCGFLNASFHIGGIFSLSFCKGGCTTCCTAAATGCCTCATTCCACATTGGGGGCATATTCAGTCTCTCTTTCTGTAAASNLVHHFFCDVPAVMALSCSDTCCAATCTGGTACATCACTTTTTCTGTGATGTTCCAGCAGTCATGGCTCTGTCTTGCKHTSEVILVFTSSFNIFFVLLTCTGATAAACACACTAGTGAGGTGATTCTGGTTTTTACGTCAAGCTTTAATATCTTTVIFISYLFIFITILKMHSAKRTTTGTTCTTCTAGTTATCTTTATCTCCTACTTGTTCATATTCATCACCATCTTGAAGHQKALSTCASHFTAVSVFYGTATGCATTCAGCTAAGAGACACCAAAAAGCATTGTCCACCTGTGCCTCTCACTTCACTVIFIYLQPSSSHSMDTDKMASGCAGTCTCCGTCTTCTATGGGACAGTAATCTTCATCTACTTGCAGCCCAGCTCCAGCVFYAMIIPMLNPVVYSLRNRECACTCCATGGACACAGACAAAATGGCATCTGTGTTCTATGCTATGATCATCCCCATGVQNAFKKVLRRQKFLCTGAACCCTGTGGTCTACAGCCTGAGGAACAGAGAAGTCCAGAATGCATTCAAGAAAGTGTTGAGAAGGCAAAAATTTCTATAAGMAC006313_A—81/82CGGTTGCCCCTGCTGAATTCGTCCTCCTGGGCATCACAAATCGCTGGGACCTGCGTGMGMALLIRMDARLHTPMYFFL dalTGGCCCTCCTCCTGACCTGCCTGCCTGTCTACCTGGTGAGCCTGCTGGGAAACATGGANLSLLDACYSSAIGPKMLVDGCATGGCGCTGCTGATCCGCATGGATGCCCGGCTCCACACACCTATGTACTTCTTCCLLLPRATIPYTACALQMFVFATGGCCAACCTCTCCCTGCTGGATGCCTGCTATTCCTCCGCCATCGGCCCCAAGATGCGLADTECCLLAAMAYDRYVAITAGTGGACCTGCTGCTGCCCCGAGCCACCATCCCTTACACAGCCTGTGCCCTCCAGARNPLLYTTAMSQRLCLALLGATGTTTGTCTTTGCAGGTCTGGCTGATACTGAGTGTTGCTTGCTGGCAGCCATGGCCTSGLGGAVSAFVHTTLTFRLSFATGACCGCTACGTGGCCATCAGAAACCCACTTCTCTATACAACAGCTATGTCGCAGCCRSRKINSFFCDIPPLLAISCGTCTATGCCTGGCCTTGCTGGGAGCATCAGGCCTGGGTGGGGCAGTGAGTGCCTTTGSDTSLNELLLFAICGFIQTATTTCACACAACCCTCACCTTCCGTCTGAGCTTCTGCCGCTCCCGGAAGATCAATAGCTVLAITVSYGFIAGAVIHMRSVTCTTCTGCAGTATCCCTCCACTGCTGGCCATCTCGTGCAGTGACACCAGTCTCAATGEGSRRAASTGGSHLTAVAMMYAACTCCTTCTCTTCGCCATCTGTGGCTTCATCCAGACAGCCACGGTGTTAGCTATCAGTLIFMYLRPSSSYALDTDKMCGGTGTCTTATGGCTTCATCGCTGGGGCTGTGATCCACATGCGCTCGGTCGAGGGCAASVFYTPVIPSLNPLIYSLRNGTCGGCGAGCAGCCTCCACCGGTGGTTCCCACCTCACAGCCGTGGCCATGATGTACGKEVKEALRQTWSRFHCPGQGSGGACACTCATTTTCATGTACCTGCGCCCCAGCTCCAGCTATGCCCTGGACACTGACAQAGATGGCCTCTGTGTTCTATACCCCGGTCATCCCGTCTCTCAACCCACTCATCTACAGCCTCCGCAATAAGGAGGTCAAGGAGGCCCTCAGGCAGACCTGGAGCCGATTCCACTGTCCAGGGCAGGGGTCCCAGTGATTGGTCCAGGGAGGCTGGGTAGGTCTGACTATGAGGGGATGAGGAAGCG56385_0183/84CAGAGGTGACTGAATTCATCCTTGTGGGGTTAACTGATGACCCAGAACTGCAGATCCEVTEFILVGLTDDPELQIPLFCACTCTTCATAGTCTTCCTTTTCATCTACCTCATCACCCTGGTTGGGAACCTGGGGAIVFLFIYLITLVGNLGMIELITGATTGAATTGATTCTACTGGACTCCTGTCTCCACACCCCCATGTACTTCTTCCTCALLDSCLHTPMYFFLSNLSLVDGTAACCTCTCCCTGGTGGACTTTGGTTATTCCTCAGCTGTCACTCCCAAGGTGATGGFGYSSAVTPKVMVGFLTGDKFTGGGGTTTCTCACAGGAGACAAATTCATATTATATAATGCTTGTGCCACACAATTCTILYNACATQFFFFVAFITAESTCTTCTTTGTAGCCTTTATCACTGCAGAAAGTTTCCTCCTGGCATCAATGGCCTATGFLLASMAYDRYAALCKPLHYTACCGCTATGCAGCATTGTGTAAACCCCTGCATTACACCACCACCATGACAACAAATGTTMTTNVCARLAIGSYICGFLTATGTGCTCGCCTGGCCATAGGCTCCTACATCTGTGGTTTCCTGAATGCATCCATTCNASIHTGNTFRLSFCRSNVVEATACTGGGAACACTTTCAGGCTCTCCTTCTGTAGATCCAATGTAGTTGAACACTTTTHFFCDAPPLLTLSCSDNYISETCTGTGATGCTCCTCCTCTCTTGACTCTCTCATGTTCAGACAACTACATCAGTGAGAMVIFFVVGFNDLFSILVILISTGGTTATTTTTTTTGTGGTGGGATTCAATGACCTCTTTTCTATCCTGGTAATCTTGAYLFIFITIMKMRSPEGRQKAFTCTCCTACTTATTTATATTTATCACCATCATGAAGATGCGCTCACCTGAAGGACGCCSTCASHLTAVSIFYGTGIFMYAGAAGGCCTTTTCTACTTGTGCTTCCCACCTTACTGCAGTTTCCATCTTTTATGGGALRPNSSHFMGTDKMASVFYAICAGGAATCTTTATGTACTTACGACCTAACTCCAGCCATTTCATGGGCACAGACAAAAVIPMLNPLVYSLRNKEVKSAFTGGCATCTGTGTTCTATGCCATAGTCATTCCCATGTTGAATCCACTGGTCTACAGCCKKTVGKAKASIGFIFTGAGGAACAAAGAGGTTAAGAGTGCCTTTAAAAAGACTGTAGGGAAGGCAAAGGCCTCTATAGGATTCATATTTTAATTATAGMAC022882_D85/86GTTTTCTGATGAACCTGGATGGAACAACACAATCTAACAACGGTGAATGAATTCATTMEQHNLTTVNEFILTGITDIACTTACGGGAATCACAGATATCGCTGAGCTGCAGGCACCATTATTTGCATTGTTCCTCELQAPLFALFLMIYVISVMGNATGATCTATGTGATCTCAGTGATGGGCAATTTGGGCATGATTGTCCTCACCAAGTTGLGMIVLTKLDSRLQTPMYFFLGACTCCAGGTTGCAAACCCCTATGTACTTTTTTCTCAGACATCTGGCTTTCATGGATRHLAFMDLGYSTTVGPKMLVNCTTGGTTATTCAACAACTGTGGGACCCAAAATGTTAGTAAATTTTGTTGTGGATAAGFVVDKNIISYYFCATQLAFFLAATATAATTTCTTATTATTTTTGTGCAACACAGCTAGCTTTCTTTCTTGTGTTCATTVFIGSELFILSAMSYDLYVAIGGTAGTGAACTTTTTATTCTCTCAGCCATGTCCTACGACCTCTATGTGGCCATCTGTCNPLLYTVIMSRRVCQVLVAIAACCCTCTGCTATACACAGTAATCATGTCACGAAGGGTATGTCAGGTGCTGGTAGCAPYLYCTFISLLVTIKIFTLSFATCCCTTACCTCTATTGCACATTCATTTCTCTTCTAGTCACCATAAAGATTTTTACTCGYNVISHFYCDSLPLLPLLCTTATCCTTCTGTGGCTACAACGTCATTAGTCATTTCTACTGTGACAGTCTCCCTTTGSNTHEIELIILIFAAIDLISSTTACCTTTGCTTTGTTCAAATACACATGAAATTGAATTGATAATTCTGATCTTTGCALLIVLLSYLLILVAILRMNSAGCTATTGATTTGATTTCATCTCTTCTGATAGTTCTTTTATCTTACCTGCTCATCCTTGRQKAFSTCGAHLTVVIVFYGGTAGCCATTCTCAGGATGAATTCTGCTGGCAGACAAAAGGCTTTTTCTACCTGTGGATLLFMYVQPKSSHSFDTDKVAGCCCACCTGACAGTGGTCATAGTGTTCTATGGGACTTTGCTTTTCATGTACGTGCAGSIFYTLVIPMLNPLIYSLRNKCCCAAGTCCAGTCATTCCTTTGACACTGATAAAGTGGCTTCCATATTTTACACCCTGDVKYALRRTWNNLCNIFVGTTATCCCCATGTTGAATCCCTTGATCTATAGTTTACGAAACAAAGATGTAAAATATGCCCTACGAAGGACATGGAATAACTTATGTAATATTTTTGTTTAAATTTTCG56755-0187/88GTTTTCTGATGAACCTGGATGGAACAACACAATCTAACAACGGTGAATGAATTCATTMEQHNLTTVNEFILTGITDIACTTACGGGAATCACAGATATCGCTGAGCTGCAGGCACCATTATTTGCATTGTTCCTCELQAPLFALFLMIYVISVMGNATGATCTATGTGATCTCAGTGATGGGCAATTTGGGCATGATTGTCCTCACCAAGTTGLGMIVLTKLDSRLQTPMYFFLGACTCCAGGTTGCAAACCCCTATGTACTTTTTTCTCAGACATCTGGCTTTCATGGATRHLAFMDLGYSTTVGPKMLVNCTTGGTTATTCAACAACTGTGGGACCCAAAATGTTAGTAAATTTTGTTGTGGATAAGFVVDKNIISYYFCATQLAFFLAATATAATTTCTTATTATTTTTGTGCAACACAGCTAGCTTTCTTTCTTGTGTTCATTVFIGSELFILSAMSYDLYVAIGGTAGTGAACTTTTTATTCTCTCAGCCATGTCCTACGACCTCTATGTGGCCATCTGTCNPLLYTVIMSRRVCQVLVAIAACCCTCTGCTATACACAGTAATCATGTCACGAAGGGTATGTCAGGTGCTGGTAGCAPYLYCTFISLLVTIKIFTLSFATCCCTTACCTCTATTGCACATTCATTTCTCTTCTAGTCACCATAAAGATTTTTACTCGYNVISHFYCDSLPLLPLLCTTATCCTTCTGTGGCTACAACGTCATTAGTCATTTCTACTGTGACAGTCTCCCTTTGSNTHEIELIILIFAAIDLISSTTACCTTTGCTTTGTTCAAATACACATGAAATTGAATTGATAATTCTGATCTTTGCALLIVLLSYLLILVAILRMNSAGCTATTGATTTGATTTCATCTCTTCTGATAGTTCTTTTATCTTACCTGCTCATCCTTGRQKAFSTCGAHLTVVIVFYGGTAGCCATTCTCAGGATGAATTCTGCTGGCAGACAAAAGGCTTTTTCTACCTGTGGATLLFMYVQPKSSHSFDTDKVAGCCCACCTGACAGTGGTCATAGTGTTCTATGGGACTTTGCTTTTCATGTACGTGCAGSIFYTLVIPMLNPLIYSLRNKCCCAAGTCCAGTCATTCCTTTGACACTGATAAAGTGGCTTCCATATTTTACACCCTGDVKYALRRTWNNLCNIFVGTTATCCCCATGTTGAATCCCTTGATCTATAGTTTACGAAACAAAGATGTAAAATATGCCCTACGAAGGACATGGAATAACTTATGTAATATTTTTGTTTAAATTTTCG56820-0189/90TCAAAATCTCCAATAGCTCCAAATTCCAGGTCTCTGAGTTCATCCTGCTGGGATTCCKISNSSKFQVSEFILLGFPGICGGGCATTCACAGCTGGCAACACTGGCTATCTCTGCCCCTGGCACTACTGTATCTCTHSWQHWLSLPLALLYLSALAACAGCACTTGCTGCAAACACCCTCATCCTCATCATCATCTGGCAGAACCCTTCTTTACNTLILIIIWQNPSLQQPMYIFAGCAGCCCATGTATATTTTCCTTGGCATCCTCTGTATGGTAGACATGGGTCTGGCCALGILCMVDMGLATTIIPKILACTACTATCATCCCTAAGATCCTGGCCATCTTCTGGTTTGATGCCAAGGTTATTAGCCIFWFDAKVISLPERFAQIYAITCCCTGAGCGCTTTGCTCAGATTTATGCCATTCACTTCTTTGTGGGCATGGAGTCTGHFFVGMESGILLCMAFDRYVAGTATCCTCCTCTGCATGGCTTTTGATAGATATGTGGCTATTTGTCACCCTCTTCGCTICHPLRYPSIVTSSLILKATLATCCATCAATTGTCACCAGTTCCTTAATCTTAAAAGCTACCCTGTTCATGGTGCTGAFMVLRNGLFVTPVPVLAAQRDGAAATGGCTTATTTGTCACTCCAGTGCCTGTGCTTGCAGCACAGCGTGATTATTGCTYCSKNEIEHCLCSNLGVTSLACCAAGAATGAAATTGAACACTGCCTGTGCTCTAACCTTGGGGTCACAAGCCTGGCTTCDDRRPNSICQLVLAWLGMGSGTGATGACAGGAGGCCAAACAGCATTTGCCAGTTGGTTCTGGCATGGCTTGGAATGGDLSLIILSYILILYSVLRLNSGGAGTGATCTAAGTCTTATTATACTGTCATATATTTTGATTCTGTACTCTGTACTTAAEAAAKALSTCSSHLTLILFFGACTGAACTCAGCTGAAGCTGCAGCCAAGGCCCTGAGCACTTGTAGTTCACATCTCAYTIVVVISVTHLTEMKATLIPCCCTCATCCTTTTCTTTTACACTATTGTTGTAGTGATTTCAGTGACTCATCTGACAGVLLNVLHNIIPPSLNPTVYALAGATGAAGGCTACTTTGATTCCAGTTCTACTTAATGTGTTGCACAACATCATCCCCCQTKELRAAFQKVLFALTKEIRCTTCCCTCAACCCTACAGTTTATGCACTTCAGACCAAAGAACTTAGGGCAGCCTTCCAAAAGGTGCTGTTTGCCCTTACAAAAGAAATAAGATCTTAGAGACCTTCTCCACG56799-0191/92ACACTGAATAAAACAGACCTAATACCAGCTTCATTTATTCTGAATGGAGTCCCAGGATLNKTDLIPASFILNGVPGLECTGGAAGACACACAACTCTGGATTTCCTTCCCATTCTGCTCTATGTATGTTGTGGCTDTQLWISFPFCSMYVVAMVGNATGGTAGGGAATTGTGGACTCCTCTACCTCATTCACTATGAGGATGCCCTGCACAAACGLLYLIHYEDALHKPMYYFLCCCATGTACTACTTCTTGGCCATGCTTTCCTTTACTGACCTTGTTATGTGCTCTAGTAMLSFTDLVMCSSTIPKALCIACAATCCCTAAAGCCCTCTGCATCTTCTGGTTTCATCTCAAGGACATTGGATTTGATFWFHLKDIGFDECLVQMFFIHGAATGCCTTGTCCAGATGTTCTTCATCCACACCTTCACAGGGATGGAGTCTGGGGTGTFTGMESGVLMLMALDRYVAICTTATGCTTATGGCCCTGGATCGCTATGTGGCCATCTGCTACCCCTTACGCTATTCACYPLRYSTILTNPVIAKVGTAACTATCCTCACCAATCCTGTAATTGCAAAGGTTGGGACTGCCACCTTCCTGAGAGGGTFLRGVLLIIPFTFLTKRLPYGTATTACTCATTATTCCCTTTACTTTCCTCACCAAGCGCCTGCCCTACTGCAGAGGCCRGNILPHTYCDHMSVAKLSCAATATACTTCCCCATACCTACTGTGACCACATGTCTGTAGCCAAATTGTCCTGTGGTGNVKVNAIYGLMVALLIGGFDAATGTCAAGGTCAATGCCATCTATGGTCTGATGGTTGCCCTCCTGATTGGGGGCTTTILCITISYTMILRAVVSLSSAGACATACTGTGTATCACCATCTCCTATACCATGATTCTCCGGGCAGTGGTCAGCCTCDARQKAFNTCTAHICAIVFSYTCCTCAGCAGATGCTCGGCAGAAGGCCTTTAATACCTGCACTGCCCACATTTGTGCCTPAFFSFFSHRFGEHIIPPSCATTGTTTTCTCCTATACTCCAGCTTTCTTCTCCTTCTTTTCCCACCGCTTTGGGGAAHIIVANIYLLLPPTMNPIVYGCACATAATCCCCCCTTCTTGCCACATCATTGTAGCCAATATTTATCTGCTCCTACCAVKTKQIRDCVIRILSGSKDTKCCCACTATGAACCCTATTGTCTATGGGGTGAAAACCAAACAGATACGAGACTGTGTCSYSMATAAGGATCCTTTCAGGTTCTAAGGATACCAAATCCTACAGCATGTGAATGAACACTTGCG56929-0193/94AATGAAGAGAAAGAACTTCACAGAAGTGTCAGAATTCATTTTCTTGGGATTTTCTAGMKRKNFTEVSEFIFLGFSSFGCTTTGGAAAGCATCAGATAACCCTCTTTGTGGTTTTCCTAACTGTCTACATTTTAACKHQITLFVVFLTVYILTLVANTCTGGTTGCTAACATCATCATTGTGACTATCATCTGCATTGACCATCATCTCCACACIIIVTIICIDHHLHTPMYFFLTCCCATGTATTTCTTCCTAAGCATGCTGGCTAGTTCAGAGACGGTGTACACACTGGTSMLASSETVYTLVIVPRMLLSCATTGTGCCACGAATGCTTTTGAGCCTCATTTTTCATAACCAACCTATCTCCTTGGCLIFHNQPISLAGCATQMFFFVAGGCTGTGCTACACAAATGTTCTTTTTTGTTATCTTGGCCACTAATAATTGCTTCCTILATNNCFLLTAMGYDRYVAIGCTTACTGCAATGGGGTATGACCGCTATGTGGCCATCTGCAGACCCCTGAGATACACCRPLRYTVIMSKGLCAQLVCGTGTCATCATGAGCAAGGGACTATGTGCCCAGCTGGTGTGTGGGTCCTTTGGCATTGGSFGIGLTMAVLHVTAMFNLPFTCTGACTATGGCAGTTCTCCATGTGACAGCCATGTTCAATTTGCCGTTCTGTGGCACCGTVVDHFFCDIYPVMKLSCIAGTGGTAGACCACTTCTTTTGTGACATTTACCCAGTCATGAAACTTTCTTGCATTGADTTINEIINYGVSSFVIFVPITACCACTATCAATGAGATAATAAATTATGGTGTAAGTTCATTTGTGATTTTTGTGCCGLIFISYVLVISSILQIASAECATAGGCCTGATATTTATCTCCTATGTCCTTGTCATCTCTTCCATCCTTCAAATTGCGWKKTFATCVSHLTVVIVHCGCTCAGCTGAGGGCTGGAAGAAGACCTTTGCCACCTGTGTCTCCCACCTCACTGTGGTCASIAYLKPKSESSIEKDLVLTATTGTCCACTGTGGCTGTGCCTCCATTGCCTACCTCAAGCCGAAGTCAGAAAGTTCSVTYTIITPLLNPVVYSLRNKAATAGAAAAAGACCTTGTTCTCTCAGTGACGTACACCATCATCACTCCCTTGCTGAAEVKDALCRVVGRNISCCCTGTTGTTTACAGTCTGAGAAACAAGGAGGTAAAGGATGCCCTATGCAGAGTTGTGGGCAGAAATATTTCTTAATGGATTGGATTATTTCATTAAGAGACCATTAGCCCAGMAL049739_B95/96GCCATGGTTAACCAAAGCTCCCCCATGGGCTTCCTCCTTCTGGGCTTCTCTGAACACMVNQSSPMGFLLLGFSEHPALCCAGCACTGGAAAGGACTCTCTTTGTGGTTGTCTTCACTTCCTACCTCTTGACCCTGERTLFVVVFTSYLLTLVGNTLGTGGGCAACACACTCATCATCCTGCTGTCTGTACTGTACCCCAGGCTCCACTCTCCAIILLSVLYPRLHSPMYFFLSDATGTACTTTTTCCTCTCTGACCTCTCCTTCTTGGACCTCTGCTTTACCACAAGTTGTLSFLDLCFTTSCVPQMLVNLWGTCCCCCAGATGCTGGTCAACCTCTGGGGCCCAAAGAAGACCATCAGCTTCCTGGGAGPKKTISFLGCSVQLFIFLSLTGCTCTGTCCAGCTCTTCATCTTCCTGTCCCTGGGGACCACTGAGTGCATCCTCCTGGTTECILLTVMAFDRYVAVCQACAGTGATGGCCTTTGACCGATACGTGGCTGTCTGCCAGCCCCTCCACTATGCCACCPLHYATIIHPRLCWQLASVAWATCATCCACCCCCGCCTGTGCTGGCAGCTGGCATCTGTGGCCTGGGTTATGAGTCTGVMSLVQSIVQTPSTLHLPFCPGTTCAATCGATAGTCCAGACACCATCCACCCTCCACTTGCCCTTCTGTCCCCACCAGHQQIDDFLCEVPSLIRLSCGDCAGATAGATGACTTTTTATGTGAGGTCCCATCTCTGATTCGACTCTCCTGTGGAGATTSYNEIQLAVSSVIFVVVPLSACCTCCTACAATGAAATCCAGTTGGCTGTGTCCAGTGTCATCTTCGTGGTTGTGCCTLILASYGATAQAVLRINSATACTCAGCCTCATCCTTGCCTCTTATGGAGCCACTGCCCAGGCAGTGCTGAGGATTAACWRKAFGTCSSHLTVVTLFYSSTCTGCCACAGCATGGAGAAAGGCCTTTGGGACCTGCTCCTCCCATCTCACTGTGGTCVIAVYLQPKNPYAQGRGKFFGACCCTCTTCTACAGCTCAGTCATTGCTGTCTACCTCCAGCCCAAAAATCCGTATGCCLFYAVGTPSLNPLVYTLRNKECAAGGGAGGGGCAAGTTCTTTGGTCTCTTCTATGCAGTGGGCACTCCTTCACTTAACIKRALRRLLGKERDSRESWRACCTCTCGTATACACCCTGAGGAACAAGGAGATAAAGCGAGCACTCAGGAGGTTACTAAGGGAAGGAAAGAGACTCCAGGGAAAGCTGGAGAGCTGCTTAATATACTTTCGAAACG57376-0197/98GCCATGGTTAACCAAAGCTCCCCCATGGGCTTCCTCCTTCTGGGCTTCTCTGAACACMVNQSSPMGFLLLGFSEHPALCCAGCACTGGAAAGGACTCTCTTTGTGGTTGTCTTCACTTCCTACCTCTTGACCCTGERTLFVVVFTSYLLTLVGNTLGTGGGCAACACACTCATCATCCTGCTGTCTGTACTGTACCCCAGGCTCCACTCTCCAIILLSVLYPRLHSPMYFFLSDATGTACTTTTTCCTCTCTGACCTCTCCTTCTTGGACCTCTGCTTTACCACAAGTTGTLSFLDLCFTTSCVPQMLVNLWGTCCCCCAGATGCTGGTCAACCTCTGGGGCCCAAAGAAGACCATCAGCTTCCTGGGAGPKKTISFLGCSVQLFIFLSLTGCTCTGTCCAGCTCTTCATCTTCCTGTCCCTGGGGACCACTGAGTGCATCCTCCTGGTTECILLTVMAFDRYVAVCQACAGTGATGGCCTTTGACCGATACGTGGCTGTCTGCCAGCCCCTCCACTATGCCACCPLHYATIIHPRLCWQLASVAWATCATCCACCCCCGCCTGTGCTGGCAGCTGGCATCTGTGGCCTGGGTTATGAGTCTGVMSLVQSIVQTPSTLHLPFCPGTTCAATCGATAGTCCAGACACCATCCACCCTCCACTTGCCCTTCTGTCCCCACCAGHQQIDDFLCEVPSLIRLSCGDCAGATAGATGACTTTTTATGTGAGGTCCCATCTCTGATTCGACTCTCCTGTGGAGATTSYNEIQLAVSSVIFVVVPLSACCTCCTACAATGAAATCCAGTTGGCTGTGTCCAGTGTCATCTTCGTGGTTGTGCCTLILASYGATAQAVLRINSATACTCAGCCTCATCCTTGCCTCTTATGGAGCCACTGCCCAGGCAGTGCTGAGGATTAACWRKAFGTCSSHLTVVTLFYSSTCTGCCACAGCATGGAGAAAGGCCTTTGGGACCTGCTCCTCCCATCTCACTGTGGTCVIAVYLQPKNPYAQGRGKFFGACCCTCTTCTACAGCTCAGTCATTGCTGTCTACCTCCAGCCCAAAAATCCGTATGCCLFYAVGTPSLNPLVYTLRNKECAAGGGAGGGGCAAGTTCTTTGGTCTCTTCTATGCAGTGGGCACTCCTTCACTTAACIKRALRRLLGKERDSRESWRACCTCTCGTATACACCCTGAGGAACAAGGAGATAAAGCGAGCACTCAGGAGGTTACTAAGGGAAGGAAAGAGACTCCAGGGAAAGCTGGAGAGCTGCTTAATATACTTTCGAAACG59729-01 99/100ACATGGAGACAAAGAATTATAGCAGCAGCACCTCAGGCTTCATCCTCCTGGGCCTCTMEKNYSSSTSGFILLGLSSNCTTCCAACCCTAAGCTGCAGAAACCTCTCTTTGCCATCTTCCTCATCATGTACCTACPKLQKPLFAIFLIMYLLTAVGTCACTGCGGTGGGGAATGTGCTCATCATCCTGGCCATCTACTCTGACCCCAGGCTCCNVLIILAIYSDPRLHTPMYFFACACCCCTATGTACTTTTTTCTCAGCAACTTGTCTTTCATGGATATCTGCTTCACAALSNLSFMDICFTTVIVPKMLVCAGTCATAGTGCCTAAGATGCTGGTGAATTTTCTATCAGAGACAAAGATTATCTCTTNFLSETKIISYVGCLIQMYFFATGTGGGCTGCCTGATCCAGATGTACTTCTTCATGGCATTTGGGAACACTGACAGCTMAFGNTDSYLLASMAIDRLVAACCTGCTGGCCTCTATGGCCATCGACCGGCTGGTGGCCATCTGCAACCCCTTACACTICNPLHYDVVMKPWHCLLMLLATGATGTGGTTATGAAACCATGGCATTGCCTACTCATGCTATTGGGTTCTTGCAGCAGSCSISHLHSLFRVLLMSRLSTCTCCCACCTACATTCCCTGTTCCGCGTGCTACTTATGTCTCGCTTGTCTTTCTGTGFCASHIIKHFFCDTQPVLKLSCCTCTCACATCATTAAGCACTTTTTCTGTGACACCCAGCCTGTGCTAAAGCTCTCCTCSDTSSSQMVVMTETVALIVTGCTCTGACACATCCTCCAGCCAGATGGTGGTGATGACTGAGACCTTAGCTGTCATTGPFLCTIFSYLQIIVTVLRIPSTGACCCCCTTCCTGTGTACCATCTTCTCCTACCTGCAAATCATCGTCACTGTGCTCAAARKWKAFSTCGSHLTVVVLFGAATCCCCTCTGCAGCCAGGAAGTGGAAGGCCTTCTCTACCTGTGGCTCCCACCTCAYGSVIYVYFRPLSMYSVMKGRCTGTAGTGGTCCTGTTCTATGGGAGTGTCATCTATGTCTATTTTAGGCCTCTGTCCAVATVMYTVVTPMLNPFIYSLRTGTACTCAGTGATGAAGGGCCGGGTAGCCACAGTTATGTACACAGTAGTGACACCCANKDMKRGLKKLRHRIYSTGCTGAACCCTTTCATCTACAGCCTGAGGAACAAAGATATGAAAAGGGGTTTGAAGAAATTAAGACACAGAATTTACTCATAGAAAGAACAAAATCG59410-01101/102CGATGCTGCTGACAGATAGAAATACACGTGGGACCACGTTCACCCTCTTGGGCTTCTMLLTDRNTRGTTFTLLGFSDYCAGATTACCCAGAACTGCAAGTCCCACTCTTCCTGGTTTTTCTGGCCATCTACAATGPELQVPLFLVFLAIYNVTVLGTCACTGTGCTAGGGAATATTGGGTTGATTGTGATCATCAAAATCAACCCCAAACTGCNIGLIVIIKINPKLHTPMYFFATACCCCCATGTACTTTTTCCTCAGCCAACTCTCCTTTGTGGATTTCTGCTATTCCTLSQLSFVDFCYSSIIAPKMLVCCATCATTGCTCCCAAGATGTTGGTGAACCTTGTTGTCAAAGACAGAACCATTTCATNLVVKDRTISFLGCVVQFFFFTTTTAGGATGCGTAGTACAATTCTTTTTCTTCTGTACCTTTGTGGTCACTGAATCCTCTFVVTESFLLAVMAYDRFVATTTTATTAGCTGTGATGGCCTATGACCGCTTCGTGGCCATTTGCAACCCTCTGCTCTICNPLLYTVDMSQKLCVLLVVACACAGTTGACATGTCCCAGAAACTCTGCGTGCTGCTGGTTGTGGGATCCTATGCCTGSYAWGVSCSLELTCSALKLCGGGGAGTCTCATGTTCCTTGGAACTGACGTGCTCTGCTTTAAAGTTATGTTTTCATGFHGFNTINHFFCEFSSLLSLSGTTTCAACACAATCAATCACTTCTTCTGTGAGTTCTCCTCACTACTCTCCCTTTCTTCSDTYINQWLLFFLATFNEISGCTCTGATACTTACATCAACCAGTGGCTGCTATTCTTTCTTGCCACCTTTAATGAAATLLIVLTSYAFIVVTILKMRSTCAGCACACTACTCATCGTTCTCACATCTTATGCGTTCATTGTTGTAACCATCCTCAVSGRRKAFSTCASHLTAITIFAGATGCGTTCAGTCAGTGGGCGCCGCAAAGCCTTCTCCACCTGTGCCTCCCACCTGAHGTILFLYCVPNSKNSRHTVKCTGCCATCACCATCTTCCATGGCACCATCCTCTTCCTTTACTGTGTGCCCAACTCCAVASVFYTVVIPMLNPLIYSLRAAAACTCCAGGCACACAGTCAAAGTGGCCTCTGTGTTTTACACCGTGGTGATCCCCANKDVKDTVTEILDTKVFSYTGTTGAATCCCCTGATCTACAGTCTGAGAAATAAAGATGTCAAGGATACAGTCACCGAGATACTGGACACCAAAGTCTTCTCTTACTGAGCCTCG59400-01103/104GCCATGAAACTATTAAATCAATCTCAAGTGTCAGAATTCATTTTGCTGGGACTGACCMKLLNQSQVSEFILLGLTSSQAGCTCCCAGGATGTAGAGTTTCTTCTCTTTGCCCTCTTCTCGGTTATCTATGTGGTCDVEFLLFALFSVIYVVTVLGNACAGTTTTGGGTAACCTTCTTATTATAGTCACAGTGTTTAACACCCCTAACCTGAATLLIIVTVFNTPNLNTPMYFLLACTCCCATGTATTTTCTCCTTGGTAATCTCTCTTTTGTAGATATGACCCTTGCTTCTGNLSFVDMTLASFATPKVILNTTTGCCACCCCTAAGGTGATTCTGAACTTGTTAAAAAAGCAGAAGGTAATTTCTTTTLLKKQKVISFAGCFTQIFLLHGCTGGGTGCTTCACTCAGATATTTCTCCTTCACTTACTGGGTGGGGTTGAAATGGTALLGGVEMVLLVSMAFDRYVAICTGTTGGTCTCCATGGCTTTTGACAGATATGTGGCCATTTGTAAGCCCCTACACTACCKPLHYMTIMNKKVCVLLVVTATGACCATCATGAACAAGAAGGTATGTGTTTTGCTTGTAGTGACCTCATGGCTCTTGSWLLGLLHSGFQIPFAVNLPFGGTCTCCTTCACTCAGGGTTTCAGATACCATTTGCTGTGAACTTGCCCTTTTGTGGTCGPNVVDSIFCDLPLVIKLACCCCAATGTGGTAGACAGCATTTTTTGTGACCTCCCTTTGGTTATTAAGCTTGCCTGTIDIYFVQVVIVANSGIISLSCATAGACATATATTTTGTACAGGTAGTCATTGTTGCCAACAGTGGCATAATCTCCCTGFIILLISYSLILITIKNHSPTAGCTGTTTCATTATTTTGCTTATCTCCTACAGTCTGATCCTCATAACCATTAAGAACGQSKARSTLTAHITVVILFFGCACTCTCCTACTGGGCAATCTAAAGCCCGTTCCACTTTGACTGCTCACATCACAGTGPCIFIYIWPFGNHSVDKFLAVGTGATTCTCTTCTTTGGCCCATGCATCTTTATCTACATTTGGCCCTTCGGCAACCACFYTIITPILNPIIYTLRNKEMTCTGTAGATAAGTTCCTTGCTGTGTTTTATACCATCATCACTCCTATCTTGAATCCAKISMKKLWRAFVNSREDTATTATCTATACTCTGAGAAACAAAGAAATGAAGATATCCATGAAAAAACTCTGGAGAGCTTTTGTGAATTCTAGAGAAGATACTTAGATTAAAAATATAATGCG110254-01105/106AGAAGATGACCATGACAACGGAGAACCCCAACCAGACTGTGGTGAGCCACTTCTTCCKMTMTTENPNTQVVSHFFLEGTGGAGGGTTTGAGGTACACCGCTAAACATTCTAGCCTCTTCTTCCTCCTCTTCCTCCLRYTAKHSSLFFLLFLLIYSITCATCTACAGCATCACTGTGGCTGGGAATCTCCTCATCCTCCTAACTGTGGGCTCTGTVAGNLLILLTVGSDSHLSLPACTCTCACCTCAGCTTACCCATGTACCACTTCCTGGGGCACCTCTCCTTCCTGGATGMYHFLGHLSFLDACLSTVTVPCCTGTTTGTCTACAGTGACAGTGCCCAAGGTCATGGCAGGCCTGCTGACTCTGGATGKVMAGLLTLDGKVISFEGCAVGGAAGGTGATCTCCTTTGAGGGCTGTGCCGTACAGCTTTATTGCTTCCACTTTCTGGQLYCFHFLASTECFLYTVMAYCCAGCACTGAGTGCTTCCTGTACACAGTCATGGCCTATGACCGCTATCTGGCTATCTDRYLAICQPLHYPVAMNRRMCGTCAACCCCTGCACTACCCAGTGGCCATGAACAGAAGGATGTGTGCAGAAATGGCTGAEMAGITWAIGATHAAIHTSLGAATCACCTGGGCCATAGGTGCCACGCACGCTGCAATCCACACCTCCCTCACCTTCCTFRLLYCGPCHIAYFFCDIPPGCCTGCTCTACTGTGGGCCTTGCCACATTGCCTACTTCTTCTGCGACATACCCCCTGVLKLACTDTTINELVMLASIGTCCTAAAGCTCGCCTGTACAGACACCACCATTAATGAGCTAGTCATGCTTGCCAGCAIVAAGCLILIVISYIFIVAAVTTGGCATCGTGGCTGCAGGCTGCCTCATCCTCATCGTTATTTCCTACATCTTCATCGLRIRTAQGRQRAFSPCTAQLTTGGCAGCTGTGTTGCGCATCCGCACAGCCCAGGGCCGGCAGCGGGCCTTCTCCCCCTGVLLYYVPPVCIYLQPRSSEAGCACTGCCCAGCTCACTGGGGTGCTCCTGTACTACGTGCCACCTGTCTGTATCTACCGAGAPAVFYTIVTPMLNPFIYTGCAGCCTCGCTCCAGTGAGGCAGGAGCTGGGGCCCCTGCTGTCTTCTACACAATCGTLRNKEVKHALQRLLCSSFRETAACTCCAATGCTCAACCCATTCATTTACACTTTGCGGAACAAGGAGGTGAAGCATGSTAGSPPPCTCTGCAAAGGCTTTTGTGCAGCAGCTTCCGAGAGTCTACAGCAGGCAGCCCACCCCCATAGTCTGTGCTATCAAAACTGMAC073079_B107/108GGCCCCATACTGTGGATCATGGCAAATCTGAGCCAGCCCTCCGAATTTGTCCTCTTGMANLSQPSEFVLLGFSSFGELGGCTTCTCCTCCTTTGGTGAGCTGCAGGCCCTTCTGTATGGCCCCTTCCTCATGCTTQALLYGPFLMLYLLAFMGNTITATCTTCTCGCCTTCATGGGAAACACCATCATCATAGTTATGGTCATAGCTGACACCIIVMVIADTHLHTPMYFFLGNCACCTACATACACCCATGTACTTCTTCCTGGGCAATTTTTCCCTGCTGGAGATCTTGFSLLEILVTMTAVPRMLSDLLGTAACCATGACTGCAGTGCCCAGGATGCTCTCAGACCTGTTGGTCCCCCACAAAGTCVPHKVITFTGCMVQFYFHFSLATTACCTTCACTGGCTGCATGGTCCAGTTCTACTTCCACTTTTCCCTGGGGTCCACCGSTSFLILTDMALDRFVAICHTCCTTCCTCATCCTGACAGACATGGCCCTTGATCGCTTTGTGGCCATCTGCCACCCAPLRYGTLMSRAMCVQLAGAAWCTGCGCTATGGCACTCTGATGAGCCGGGCTATGTGTGTCCAGCTGGCTGGGGCTGCCAAPFLAMVPTVLSRAHLDYCHTGGGCAGCTCCTTTCCTAGCCATGGTACCCACTGTCCTCTCCCGAGCTCATCTTGATGDVINHFFCDNEPLLQLSCSDTACTGCCATGGCGACGTCATCAACCACTTCTTCTGTGACAATGAACCTCTCCTGCAGTRLLEFWDFLMALTFVLSSFLTTGTCATGCTCTGACACTCGCCTGTTGGAATTCTGGGACTTTCTGATGGCCTTGACCVTLISYGYIVTTVLRIPSASSTTTGTCCTCAGCTCCTTCCTGGTGACCCTCATCTCCTATGGCTACATAGTGACCACTCQKAFSTCGSHLTLVFIGYSSGTGCTGCGGATCCCCTCTGCCAGCAGCTGCCAGAAGGCTTTCTCCACTTGCGGGTCTTIFLYVRPGKAHSVQVRKVVACACCTCACACTGGTCTTCATCGGCTACAGTAGTACCATCTTTCTGTATGTCAGGCCTLVTSVLTPFLNPFILTFCNQTGGCAAAGCTCACTCTGTGCAAGTCAGGAAGGTCGTGGCCTTGGTGACTTCAGTTCTCVKTVLQGQMQRLKGLCKAQACCCCCTTTCTCAATCCCTTTATCCTTACCTTCTGCAATCAGACAGTTAAAACAGTGCTACAGGGGCAGATGCAGAGGCTGAAAGGCCTTTGCAAGGCACAATGATGAGGMAP000723_A109/110ATGAGACAGAATAACAATATTACAGAATTTGTCCTCCTGGGCTTTTCTCAGGATCCTMRQNNNITEFVLLGFSQDPGVGGTGTGCAAAAAGCATTATTTGTCATGTTTTTACTCACATACTTGGTGACAGTGGTGQKALFVMFLLTYLVTVVGNLLGGGAACCTGCTCATTGTGGTGGATATTATTGCCAGCCCTTCCTTGGGTTCCCCAATGIVVDIIASPSLGSPMYFFLACTATTTCTTCCTTGCCTGCCTGTCATTTATAGATGCTGCATATTCCACTACCATTTCTLSFIDAAYSTTISFKLIVGLFCCCAAGTTAATTGTAGGCTTATTCTGTGATAAAAAGACTATTTCCTTCCAAGGTTGCCDKKTISFQGCMGQLFIDHFFATGGGCCAGCTATTTATAGACCATTTCTTTGGTGGGGCTGAGGTCTTCCTTCTGGTGGGAEVFLLVVMACDRYVAICKGTGATGGCCTGTGATCGCTATGTGGCCATCTGTAAGCCACTGCACTATTTGACCATCPLHYLTIMNRQVCFLLLVVAMATGAATCGACAGGTTTGCTTCCTTCTGTTGGTGGTGGCCATGATTGGAGGTTTTGTAIGGFVHSAFQIVVYSLPFCGPCATTCTGCGTTTCAAATTGTTGTGTACAGTCTCCCTTTCTGTGGTCCCAATGTCATTNIVVHFSCDMHPLLELACTDTGTTCATTTCAGTTGTGACATGCACCCATTACTGGAACTGGCATGCACTGACACCTACYFIGLTVVVNSGAICMVIFNLTTTATAGGCCTCACTGTTGTTGTCAATAGTGGAGCAATCTGTATGGTCATTTTCAACLLISYGVILSSLKTYSQEKRGCTTCTGTTAATCTCCTATGGAGTCATCCTAAGCTCCCTTAAAACTTACAGTCAGGAAKALSTCSSGSTVVVLFFVPCIAAGAGGGGTAAAGCCTTGTCTACCTGCAGCTCCGGCAGTACCGTTGTTGTCCTCTTTFIYVRPVSNFPTDKRMTVFYTTTTGTACCCTGTATTTTCATATATGTTAGACCTGTTTCAAACTTTCCTACTGATAAGIITHMLSPLIYTLRNSEMRNATTCATGACTGTGTTTTATACCATTATCACACACATGCTGAGTCCTTTAATATATACGIEKLLGKKLTIFIIGGVSVLMTTGAGAAATTCAGAGATGAGAAATGCTATAGAAAAACTCTTGGGTAAAAAGTTAACTATATTTATTATAGGAGGAGTGTCCGTCCTCATGTAGGMAP001521_A111/112ACTTTTACTAGCCATGACACTAGGAAACAGCACTGAAGTCACTGAATTCTATCTTCTMTLGNSTEVTEFYLLGFGAQHGGGATTTGGTGCCCAGCATGAGTTTTGGTGTATCCTCTTCATTGTATTCCTTCTCATEFWCILFIVFLLIYVTSIMGNCTATGTGACCTCCATAATGGGTAATAGTGGAATAATCTTACTCATCAACACAGATTCSGIILLINTDSRFQTLTYFFLCAGATTTCAAACACTCACGTACTTTTTTCTACAACATTTGGCTTTTGTTGATATCTGQHLAFVDICYTSAITPKMLQSTTACACTTCTGCTATCACTCCCAAGATGCTCCAAAGCTTCACAGAAGAAAAGAATTTFTEEKNLILFQGCVIQFLVYAGATATTATTTCAGGGCTGTGTGATACAATTCTTAGTTTATGCAACATTTGCAACCAGTFATSDCYLLAMMAVDPYVAITGACTGTTATCTCCTGGCTATGATGGCAGTGGATCCTTATGTTGCCATCTGTAAGCCCKPLHYTVIMSRTVCIRLVAGCCTTCACTATACTGTAATCATGTCCCGAACAGTCTGCATCCGTTTGGTAGCTGGTTCSYIMGSINASVQTGFTCSLSFATACATCATGGGCTCAATAAATGCCTCTGTACAAACAGGTTTTACATGTTCACTGTCCKSNSINHFFCDVPPILALSCCTTCTGCAAGTCCAATAGCATCAATCACTTTTTCTGTGATGTTCCCCCTATTCTTGCSNVDINIMLLVVFVGSNLIFTTCTTTCATGCTCCAATGTTGACATCAACATCATGCTACTTGTTGTCTTTGTGGGATCGLVVIFSYIYIMATILKMSSSTAACTTGATATTCACTGGGTTGGTCGTCATCTTTTCCTACATCTACATCATGGCCACAGRKKSFSTCASHLTAVTIFYCATCCTGAAAATGTCTTCTAGTGCAGGAAGGAAAAAATCCTTCTCAACATGTGCTTCGRLSYMYLQSHSNNSQENMKVCCACCTGACCGCAGTCACCATTTTCTATGGGACACTCTCTTACATGTATTTGCAGTCAFIFYGTVIPMLNPLIYSLRNTCATTCTAATAATTCCCAGGAAAATATGAAAGTGGCCTTTATATTTTATGGCACAGTKEVKEALKVIGKKLFTATTCCCATGTTAAATCCTTTAATCTATAGCTTGAGAAATAAGGAAGTAAAAGAAGCTTTAAAAGTGATAGGGAAAAAGTTATTTTAAATCAGCCCCAGMAP002826_A113/114CCATGACACTAGGAAACAGCACTGAAGTCACTGAATTCTATCTTCTGGGATTTGGTGMTLGNSTEVTEFYLLGFGAQHCCCAGCATGAGTTTTGGTGTATCCTCTTCATTGTATTCCTTCTCATCTATGTGACCTEFWCILFIVFLLIYVTSIMGNCCATAATGGGTAATAGTGGAATAATCTTACTCATCAACACAGATTCCAGATTTCAAASGIILLINTDSTFQTLTYFFLCACTCACGTACTTTTTTCTACAACATTTGGCTTTTGTTGATATCTGTTACACTTCTGQHLAFVDICYTSAITPKMLQSCTATCACTCCCAAGATGCTCCAAAGCTTCACAGAAGAAAAGAATTTGATGTTATTTCFTEEKNLMLFQGCVIQFLVYAAGGGCTGTGTGATACAATTCTTAGTTTATGCAACATTTGCAACCAGTGACTGTTATCTFATSDCYLLAMMAVDPYVAITCCTGGCTATGATGGCAGTGGATCCTTATGTTGCCATCTGTAAGCCCCTTCACTATACKPLHYTVIMSRTVCIRLVAGCTGTAATCATGTCCCGAACAGTCTGCATCCGTTTGGTAGCTGGTTCATACATCATGGSYIMGSINASVQTGFTCSLSFGCTCAATAAATGCCTCTGTACAAACAGGTTTTACATGTTCACTGTCCTTCTGCAAGTCKSNSINHFFCDVPPILALSCCCAATAGCATCAATCACTTTTTCTGTGATGTTCCCCCTATTCTTGCTCTTTCATGCTSNVDINIMLLVVFVGSNLIFTCCAATGTTGACATCAACATCATGCTACTTGTTGTCTTTGTGGGATCTAACTTGATATGLVVIFSYIYIMATILKMSSSTCACTGGGTTGGTCGTCATCTTTTCCTACATCTACATCATGGCCACCATCCTGAAAAAGRKKSFSTCASHLTAVTIFYTGTCTTCTAGTGCAGGAAGGAAAAAATCCTTCTCAACATGTGCTTCCCACCTGACCGGTLSYMYLQSHSNNSQENMKVCAGTCACCATTTTCTATGGGACACTCTCTTACATGTATTTGCAGTCTCATTCTAATAAFIFYGTVIPMLNPLIYSLRNATTCCCAGGAAAATATGAAAGTGGCCTTTATATTTTATGGCACAGTTATTCCCATGTKEVKEALKVIGKKLFTAAATCCTTTAATCTATAGCTTGAGAAATAAGGAAGTAAAAGAAGCTTTAAAAGTGATAGGGAAAAAGTTATTTTAAATCAGCCCCAGM2557p19A115/116TGCATTGTGTAAAACATGGGGGATGTGAATCAGTCGGTGGCCTCAGACTTCATTCTGMGDVNQSVASDFILVGLFSHSGTGGGCCTCTTCAGTCACTCAGGATCACGCCAGCTCCTCTTCTCCCTGGTGGCTGTCGSRQLLFSLVAVMFVIGLLGNATGTTTGTCATAGGCCTTCTGGGCAACACCGTTCTTCTCTTCTTGATCCGTGTGGACTVLLFLIRVDSRLHTPMYFLLTCCCGGCTCCATACACCCATGTACTTCCTGCTCAGCCAGCTCTCCCTGTTTGACATTSQLSLFDIGCPMVTIPKMASDGGCTGTCCCATGGTCACCATCCCCAAGATGGCATCAGACTTTCTGCGGGGAGAAGGTFLRGEGATSYGGGAAQIFFLTGCCACCTCCTATGGAGGTGGTGCAGCTCAAATATTCTTCCTCACACTGATGGGTGTGLMGVAEGVLLVLMSYDRYVAVGCTGAGGGCGTCCTGTTGGTCCTCATGTCTTATGACCGTTATGTTGCTGTGTGCCAGCQPLQYPVLMRRQVCLLMMGSCCCCTGCAGTATCCTGTACTTATGAGACGCCAGGTATGTCTGCTGATGATGGGCTCCSWVVGVLNASIQTSITLHFPYTCCTGGGTGGTAGGTGTGCTCAACGCCTCCATCCAGACCTCCATCACCCTGCATTTTCASRIVDHFFCEVPALLKLSCCCCTACTGTGCCTCCCGTATTGTGGATCACTTCTTCTGTGAGGTGCCAGCCCTACTGADTCAYEMALSTSGVLILMLPAAGCTCTCCTGTGCAGATACCTGTGCCTACGAGATGGCGCTGTCCACCTCAGGGGTGLSLIATSYGHVLQAVLSMRSECTGATCCTAATGCTCCCTCTTTCCCTCATCGCCACCTCCTACGGCCACGTGTTGCAGEARHKAVTTCSSHITVVGLFYGCTGTTCTAAGCATGCGCTCAGAGGAGGCCAGACACAAGGCTGTCACCACCTGCTCCGAAVFMYMVPCAYHSPQQDNVTCGCACATCACGGTAGTGGGGCTCTTTTATGGTGCCGCCGTGTTCATGTACATGGTGVSLFYSLVTPTLNPLIYSLRNCCTTGCGCCTACCACAGTCCACAGCAGGATAACGTGGTTTCCCTCTTCTATAGCCTTPEVWMALVKVLSRAGLRQMCGTCACCCCTACACTCAACCCCCTTATCTACAGTCTGAGGAATCCGGAGGTGTGGATGGCTTTGGTCAAAGTGCTTAGCAGAGCTGGACTCAGGCAAATGTGCTGACTACATAGAAACTGCTGGM656022_A117/118ATGGAAATAGCCAATGTGAGTTCTCCAGAAGTCTTTGTCCTCCTGGGCTTCTCCACAMEIANVSSPEVFVLLGFSTRPCGACCCTCACTAGAAACTGTCCTCTTCATAGTTGTCTTGAGTTTTTACATGGTATCGSLETVLFIVVLSFYMVSILGNATCTTGGGCAATGGCATCATCATTCTGGTCTCCCATACAGATGTGCACCTCCACACAGIIILVSHTDVHLHTPMYFFLCCTATGTACTTCTTTCTTGCCAACCTCCCCTTCCTGGACATGAGCTTCACCACGAGCANLPFLDMSFTTSIVPQLLANATTGTCCCACAGCTCCTGGCTAACCTCTGGGGACCACAGAAAACCATAAGCTATGGALWGPQKTISYGGCVVQFYISHGGGTGTGTGGTCCAGTTCTATATCTCCCATTGGCTGGGGGCAACCGAGTGTGTCCTGWLGATECVLLATMSYDRYAAICTGGCCACCATGTCCTATGACCGCTACGCTGCCATCTGCAGGCCACTCCATTACACTCRPLHYTVIMHPQLCLGLALAGTCATTATGCATCCACAGCTTTGCCTTGGGCTAGCTTTGGCCTCCTGGCTGGGGGGTSWLGGLTTSMVGSTLTMLLPLCTGACCACCAGCATGGTGGGCTCCACGCTCACCATGCTCCTACCGCTGTGTGGGAACCGNNCIDHFFCEMPLIMQLACAATTGCATCGACCACTTCTTTTGCGAGATGCCCCTCATTATGCAACTGGCTTGTGTGVDTSLNEMEMYLASFVFVVLPGATACCAGCCTCAATGAGATGGAGATGTACCTGGCCAGCTTTGTCTTTGTTGTCCTGLGLILVSYGHIARAVLKIRSACCTCTGGGGCTCATCCTGGTCTCTTACGGCCACATTGCCCGGGCCGTGTTGAAGATCEGRRKAFNTCSSHVAVVSLFYAGGTCAGCAGAAGGGCGGAGAAAGGCATTCAACACCTGTTCTTCCCACGTGGCTGTGGSIIFMYLQPAKSTSHEQGKFGTGTCTCTGTTTTACGGGAGCATCATCTTCATGTATCTCCAGCCAGCCAAGAGCACCIALFYTVVTPALNPLIYTLRNTCCCATGAGCAGGGCAAGTTCATAGCTCTGTTCTACACCGTAGTCACTCCTGCGCTGTEVKSALRHMVAACCCACTTATTTACACCCTGAGGAACACGGAGGTGAAGAGCGCCCTCCGGCACATGGTATAGGMbA144L1_A119/120TGCTCTCCATGGAGCAAGTCAATAAGACTGTGGTGAGAGAGTTCGTCGTCCTCGGCTMEQVNKTVVREFVVLGFSSLATCTCATCCCTGGCCAGGCTGCAGCAGCTGCTCTTTGTTATCTTCCTGCTCCTCTACCRLQQLLFVIFLLLYLFTLGTNTGTTCACTCTGGGCACCAATGCAATCATCATTTCCACCATTGTGCTGGACAGAGCCCAIIISTIVLDRALHTPMYFFLTTCATACTCCCATGTACTTCTTCCTTGCCATCCTTTCTTGCTCTGAGATTTGCTATAAILSCSEICYTFVIVPKMLVDCCTTTGTCATTGTACCCAAGATGCTGGTTGACCTGCTGTCCCAGAAGAAGACCATTTLLSQKKTISFLGCAIQMFSFLCTTTCCTGGGCTGTGCCATCCAAATGTTTTCCTTCCTCTTCTTTGGCTCCTCTCACTFFGSSHSFLLAAVGYDRYMAICCTTCCTGCTGGCAGCCATGGGCTATGATCGCTATATGGCCATCTGTAACCCACTGCCNPLRYSVLMGHGVCMGLMAAGCTACTCAGTGCTCATGGGACATGGGGTGTGTATGGGACTAATGGCTGCTGCCTGTGACACGFTVSLVTTSLVFHLPFCCTGTGGCTTCACTGTCTCCCTGGTCACCACCTCCCTAGTATTTCATCTGCCCTTCCHSSNQLHHFFCDISPVLKLASACTCCTCCAACCAGCTCCATCACTTCTTCTGTGACATCTCCCCTGTCCTTAAACTGGQHSGFSQLVIFMLGVFALVIPCATCTCAGCACTCCGGCTTCAGTCAGCTGGTCATATTCATGCTTGGTGTATTTGCCTLLLILVSYIRIISAILKIPSSTGGTCATTCCTCTGCTACTTATCCTAGTCTCCTACATCCGCATCATCTCTGCCATTCVGRYKTFSTCASHLIVVTVHYTAAAAATCCCTTCCTCCGTTGGAAGATACAAGACCTTCTCCACCTGTGCCTCCCATCSCASFIYLRPKTNYTSSQDTLTCATTGTGGTAACTGTTCACTACAGTTGTGCCTCTTTCATCTACTTAAGGCCCAAGAISVSYTILTPLFNPMIYSLRNCTAATTACACTTCAAGCCAAGACACCCTAATATCTGTGTCATACACCATCCTTACCCKEFKSALRRTIGQTFYPLSCATTGTTCAATCCAATGATTTATAGTCTGAGAAATAAGGAATTCAAATCAGCCCTACGAAGAACAATCGGCCAAACTTTCTATCCTCTTAGTTAAAGGMAP000435_A121/122TATCTTGTGATTCTAAAATAACATCCATGGAGAATAATACAGAGGTGAGTGAATTCAMENNTEVSEFILLGLTNAPELTCCTGCTTGGTCTAACCAATGCCCCAGAACTACAGGTTCCCCTCTTTATCATGTTTAQVPLFIMFTLIYLITLTGNLGCCCTCATCTACCTCATCACTCTGACTGGGAACCTGGGGATGATCATATTAATCCTGCMIILILLDSHLHTPMYFFLSNTGGACTCTCATCTCCACACTCCCATGTACTTTTTTCTCAGTAACCTGTCTCTTGCAGLSLAGIGYSSAVTPKVLTGLLGCATTGGTTACTCCTCAGCTGTCACTCCAAAGGTTTTAACTGGGTTGCTTATAGAAGIEDKAISYSACAAQMFFCAVFACAAAGCCATCTCCTACAGTGCCTGTGCTGCTCAGATGTTCTTTTGTGCAGTCTTTGATVENYLLSSMAYDRYAAVCNCCACTGTGGAAAATTACCTCTTGTCCTCAATGGCCTATGACCGCTACGCAGCAGTGTPLHYTTTMTTRVCACLAIGCYGTAACCCCCTACATTATACCACCACCATGACAACACGTGTGTGTGCTTGTCTGGCTAVIGFLNASIQIGDTFRLSFCMTAGGCTGTTATGTCATTGGTTTTCTGAATGCTTCTATCCAAATTGGAGATACATTTCSNVIHHFFCDKPAVITLTCSEGCCTCTCTTTCTGCATGTCCAATGTGATTCATCACTTTTTCTGTGACAAACCAGCAGKHISELILVLISSFNVFFALLTCATTACTCTGACCTGCTCTGAGAAACACATTAGTGAGTTGATTCTTGTTCTTATATVTLISYLFILITILKRHTGKGCAAGTTTTAATGTCTTTTTTGCACTTCTTGTTACCTTGATTTCCTATCTGTTCATATYQKPLSTCGSHLIAIFLFYITTGATCACCATTCTTAAGAGGCACACAGGTAAGGGATACCAGAAGCCTTTATCTACCTVIIMYIRPSSSHSMDTDKIASGTGGTTCTCACCTCATTGCCATTTTCTTATTTTATATAACTGTCATCATCATGTACAVFYTMIIPMLSPIVYTLRNKDTACGACCAAGTTCCAGTCATTCCATGGACACAGACAAAATTGCATCTGTGTTCTACAVKNAFMKVVEKAKYSLDSVFCTATGATCATCCCCATGCTCAGTCCTATAGTCTATACCCTGAGGAACAAAGACGTGAAGAATGCATTCATGAAGGTTGTTGAGAAGGCAAAATATTCTCTAGATTCAGTCTTTTAATGATGCAAAATCATCACAATGMAC025942_B123/124ATGGAAGAGGAAAATGCAACATTGCTGACAGAGTTTGTTCTCACAGGATTTTTACATMEEENATLLTEFVLTGFLHQPCAACCTGACTGTAAAATACCGCTCTTCCTGGCATTCTTGGTAATATATCTCATCACCDCKIPLFLAFLVIYLITIMGNATCATGGGGAATCTTGGTCTAATTGTTCTCATCTGGAAAGACCCTCACCTTCATATCLGLIVLIWKDPHLHIPMYLFLCCAATGTACTTATTCCTTGGGAGTTTAGCCTTTGTGGATGCTTCGTTATCATCCACAGSLAFVDASLSSTVTPKMLINGTGACTCCGAAGATGCTGATCAACTTCTTAGCTAAGAGTAAGATGATATCTCTCTCTFLAKSKMISLSECMVQFFSLVGAATGCATGGTACAATTTTTTTCCCTTGTAACCACTGTAACCACAGAATGTTTTCTCTTVTTECFLLATMAYDRYVAITTGGCAACAATGGCATATGATCGCTATGTAGCCATTTGCAAAGCTTTACTTTATCCACKALLYPVIMTNELCIQLLVLGTCATTATGACCAATGAACTATGCATTCAGCTATTAGTCTTGTCATTTATAGGTGGCSFIGGLLHALIHEAFSFRLTFCTTCTTCATGCTTTAATCCATGAAGCTTTTTCATTCAGATTAACCTTCTGTAATTCCCNSNIIQHFYCDIIPLLKISCAACATAATACAACACTTTTACTGTGACATTATCCCATTGTTAAAGATTTCCTGTACTTDSSINFLMVFIFAGSVQVFTGATTCCTCTATTAACTTTCTAATGGTTTTTATTTTCGCAGGTTCTGTTCAAGTTTTTIGTILISYTIILFTILEKKSIACCATTGGAACTATTCTTATATCTTATACAATTATCCTCTTTACAATCTTAGAAAAGKGIRKAVSTCGAHLLSVSLYYAAGTCTATCAAAGGGATACGAAAAGCTGTCTCCACCTGTGGGGCTCATCTCTTATCTGPLTFKYLGSASPQADDQDMMGTATCTTTATACTATGGCCCCCTCACCTTCAAATATCTGGGCTCTGCATCTCCGCAAESLFYTVIVPLLNPMIYSLRNGCAGATGACCAAGATATGATGGAGTCTCTATTTTACACTGTCATAGTTCCTTTATTAKQVIASFTKMFKSNVAATCCCATGATCTACAGCCTGAGAAACAAGCAAGTAATAGCTTCATTCACAAAAATGTTCAAAAGCAATGTTTAGATCTCATACAATCTCTCTTCTCTATTTACTAAAATTGMAC055861_A125/126TCTGAGGCAATGAATGGAATGAATCACTCTGTGGTATCAGAATTTGTATTCATGGGAMNGMNHSVVSEFVFMGLTNSRCTCACCAACTCACGGGAGATTCAGCTTCTACTTTTTGTTTTCTCTTTGTTGTTCTACEIQLLLFVFSLLFYFASMMGNTTTGCGAGCATGATGGGAAACCTTGTCATTGTATTCACTGTAACCATGGATGCTCATLVIVFTVTMDAHLHSPMYFLLCTGCACTCCCCCATGTATTTCCTCCTGGCTAACCTCTCAATCATTGATATGGCATTTANLSIIDMAFCSITAPKMICDTGCTCAATTACAGCCCCTAAGATGATTTGTGATATTTTCAAGAAGCACAAGGCCATCIFKKHKAISFRGCITQIFFSHTCCTTTCGGGGATGTATTACTCAGATCTTCTTTAGCCATGCTCTTGGGGGCACTGAGALGGTEMVLLIAMAFDRYMAIATGGTGCTGCTCATAGCCATGGCCTTTGACAGATACATGGCCATATGTAAACCTCTCCKPLHYLTIMSPRMCLYFLATCACTACCTGACCATCATGAGCCCAAGAATGTGTCTATACTTTTTAGCCACTTCCTCTSSIIGLIHSLVQLVFVVDLPFATCATTGGCCTTATCCACTCATTGGTCCAATTAGTTTTTGTGGTAGATTTACCTTTTCGPNIFDSFYCDLPRLLRLACTGTGGTCCTAATATCTTTGACAGTTTTTACTGTGATCTCCCTCGGCTCCTCAGACTTTNTQELEFMVTVNSGLISVGSGCCTGTACCAACACCCAAGAACTGGAGTTCATGGTCACTGTCAATAGTGGACTCATTFVLLVISYIFILFTVWKHSSGTCTGTGGGCTCCTTTGTCTTGCTGGTAATTTCCTACATCTTCATTCTGTTCACTGTTGLAKALSTLSAHVTVVILFFGTGGAAACATTCTTCTGGTGGTCTAGCCAAGGCCCTCTCTACCCTGTCAGCTCATGTCPLMFFYTWPSPTSHLDKYLAIACTGTGGTCATCTTGTTCTTTGGGCCACTGATGTTTTTCTACACATGGCCTTCTCCCFDAFITPFLNPVIYTFRNKDMACATCACACCTGGATAAATATCTTGCTATTTTTGATGCATTTATTACTCCTTTTCTGKVAMRRLCSRLAHFTKILAATCCAGTTATCTACACATTCAGGAACAAAGACATGAAAGTGGCAATGAGGAGACTGTGCAGTCGTCTTGCGCATTTTACAAAGATTTTGTAAATGGCTTGGCTGMAC044810_B127/128CTAATGGACTGTGGCTTTATTCTGTATATACTATGTCCATAAAATCAATGCACGACTMERQNQSCVVEFILLGFSNYPTCATTACTGAAAATGGAAAGACAAAATCAAAGCTGTGTGGTTGAATTCATCCTCTTGELQGQLFVAFLVIYLVTLIGNGGCTTTTCTAACTATCCTGAGCTCCAGGGGCAGCTCTTTGTGGCTTTCCTGGTTATTAIIIVIVSLDQSLHVPMYLFLTATCTGGTGACCCTGATAGGAAATGCCATTATTATAGTCATCGTCTCCCTAGACCAGLNLSVVDLSFSAVIMPEMLVVAGCCTCCACGTTCCCATGTACCTGTTTCTCCTGAACTTATCTGTGGTGGACCTGAGTLSTEKTTISFGGCFAQMYFILTTCAGTGCAGTTATTATGCCTGAAATGCTGGTGGTCCTCTCTACTGAAAAAACTACALFGGAECFLLGAMAYDRFAAIATTTCTTTTGGGGGCTGTTTTGCACAGATGTATTTCATCCTTCTTTTTGGTGGGGCTCHPLNYQMIMNKGVFMKLIIFGAATGTTTTCTTCTGGGAGCAATGGCTTATGACCGATTTGCTGCAATTTGCCATCCTSWALGFMLGTVQTSWVSSFPFCTCAACTACCAAATGATTATGAATAAAGGAGTTTTTATGAAATTAATTATATTTTCACGLNEINHISCETPAVLELACTGGGCCTTAGGTTTTATGTTAGGTACTGTTCAAACATCATGGGTATCTAGTTTTCCCADTFLFEIYAFTGTFLIILVPTTTTGTGGCCTTAATGAAATTAACCATATATCTTGTGAAACCCCAGCAGTGTTAGAAFLLILLSYIRVLFAILKMPSTCTTGCATGTGCAGACACGTTTTTGTTTGAAATCTATGCATTCACAGGCACCTTTTTGTGRQKAFSTCAAHLTSVTLFYATTATTTTGGTTCCTTTCTTGTTGATACTCTTGTCTTACATTCGAGTTCTGTTTGCCGTASMTYLQPKSGYSPETKKVATCCTGAAGATGCCATCAACCACTGGGAGACAAAAGGCCTTTTCCACCTGTGCCGCTMSLSYSLLTPLLNLLIYSLRNCACCTCACATCTGTGACCCTATTCTATGGCACAGCCAGTATGACTTATTTACAACCCMSLSYSLLTPLLNLLIYSLRNAAATCTGGCTACTCACCGGAAACCAAGAAAGTGATGTCATTGTCTTACTCACTTCTGACACCACTGCTGAATCTGCTTATCTACAGTTTGCGAAATAGTGAGATGAAGAGGGCTTTGATGAAATTATGGCGAAGGCGAGTGGTTTTACACACAATCTGACTGMbA144L1_C129/130AGTCATGACCACCATAATTCTGGAAGTAGATAATCATACAGTGACAACACGTTTCATMTTIILEVDNHTVTTRFILLGTCTTCTGGGGTTTCCAACACGACCAGCCTTCCAGCTTCTCTTTTTCTCCATTTTCCTFPTRPAFQLLFFSIFLATYLLGGCAACCTATCTGCTGACACTGCTGGAGAATCTTCTTATCATCTTAGCTATCCACAGTLLENLLIILAIHSDGQLHKPTGATGGGCAGCTGCATAAGCCCATGTACTTCTTCTTGAGCCACCTCTCCTTCCTGGAMYFFLSHLSFLEMWYVYVISPGATGTGGTATGTCACAGTCATCAGCCCCAAGATGCTTGTTGACTTCCTCAGTCATGAKMLVDFLSHDKSISFNGCMTQCAAGAGTATTTCCTTCAATGGCTGCATGACTCAACTTTACTTTTTTGTGACCTTTGTLYFFVTFVCTEYILLAIMAFDCTGCACTGAGTACATCCTTCTTGCTATCATGGCCTTTGACCGCTATGTAGCCATTTGRYVAICNPLRYPVIMTNQLCGTAATCCACTACGCTACCCAGTCATCATGACCAACCAGCTCTGTGGCACACTGGCTGGTLAGGCWFCGLMTAMIKMVFIAGGATGCTGGTTCTGTGGACTCATGACTGCCATGATTAAGATGGTTTTTATAGCACAAQLHYCGMPQINHYFCDISPLACTTCACTACTGTGGCATGCCTCAGATCAATCACTACTTTTGTGATATCTCTCCACTLNVSCEDASQAEMVDFFLALMCCTTAACGTCTCCTGTGAGGATGCCTCACAGGCTGAGATGGTGGACTTCTTCTTGGCVIAIPLCVVVASYAAILATILCCTCATGGTCATTGCTATTCCTCTTTGTGTTGTGGTGGCATCCTACGCTGCTATCCTRIPSAQGRQKAFSTCASHLTVTGCCACCATCCTCAGGATCCCTTCTGCTCAGGGCCGCCAAAAGGCATTCTCCACCTGVILFYSMYLFTYARPKLMYAYTGCCTCCCACCTGACCGTCGTAATTCTCTTCTATTCCATGACACTTTTCACCTATGCNSNKVVSVLYTVIVPLLNPIICCGTCCCAAACTCATGTATGCCTACAATTCCAACAAAGTGGTATCTGTTCTCTACACYCLRNHEVKAALRKTIHCRGSTGTCATTGTTCCACTCCTCAACCCCATCATTTACTGTCTGAGGAACCATGAAGTAAAGPQGNGAFSSGGCAGCCCTCAGAAAGACCATACATTGCAGAGGAAGTGGGCCCCAGGGAAATGGGGCTTTCAGTAGTTAAAAAATGTATAGCG50271_01131/132TGACTGAGTTTGTTTTATTTGGCCTTTTTGAGAGCAGAGAGATGCAGCATACATGCTTEFVLFGLFESREMQHTCFVVTTGTGGTATTCTTCCTCTTTCATGTGCTCACTGTCCTGGGGAACCTTCTGGTCATCAFFLFHVLTVLGNLLVIITINATCACCATCAATGCTAGAAAGACCCTGAAGTCTCCCATGTATTTCTTCCTGAGCCAGTRKTLKSPMYFFLSQLSFADICTGTCTTTTGCTGACATATGTTATCCATCCACTACCATACCCAAGATGATTGCTGACAYPSTTIPKMIADTFVEHKIISCTTTTGTGGAGCATAAGATCATCTCCTTCAATGGCTGCATGACCCAGCTCTTTTCTGFNGCMTQLFSAHFFGGTEIFLCCCACTTCTTTGGTGGCACTGAGATCTTCCTCCTTACAGCCATGGCCTATGACCGCTLTAMAYDRYVAICRPLHYTAIATGTGGCCATCTGTAGGCCCCTGCACTACACAGCCATCATGGATTGCCGGAAGTGTGMDCRKCGLLAGASWLAGFLHSGCCTGCTAGCGGGGGCCTCCTGGTTAGCTGGCTTCCTGCATTCCATCCTGCAGACCCILQTLLTVQLPFCGPNEIDNFTCCTCACGGTTCAGCTGCCTTTTTGTGGGCCCAATGAGATAGACAACTTCTTCTGTGFCDVHPLLKLACADTYMVGLIATGTTCATCCCCTGCTCAAGTTGGCCTGTGCAGACACCTACATGGTAGGTCTCATCGVVANSGMISLAFFFILIISYVTGGTGGCCAACAGCGGTATGATTTCTTTAGCATTCTTTTTTATCCTTATCATTTCCTIILLNLRSQSSEDRRKAVSTCATGTTATCATCTTACTGAACCTAAGAAGCCAGTCATCTGAGGACCGGCGTAAGGCTGTCTCCACATGTGGCTCACACGTAATCACTGTCCTTTTGGTTCTCATGCCCCCCATGTSTTLAADKLIILFNIVMPPLLTCATGTACATTCGTCCCTCCACCACCCTGGCTGCTGACAAACTTATCATCCTCTTTANPLIYTLRNNDVKNAMRKLFRACATTGTGATGCCACCTTTGCTGAACCCTTTGATCTATACACTAAGGAACAATGATGVKRSLGEKTGAAAAATGCCATGAGGAAGCTGTTTAGGGTCAAGAGGAGCTTAGGGGAGAAGTGACATTCCAGAGAATCTCAATCCAGCTAGCG57497-01133/134ACCTATGAACAGGTCAGCAACACACATCGTGACAGAGTTTATTCTCCTGGGATTCCCMNRSATHIVTEFILLGFPGCWTGGTTGCTGGAAGATTCAGATTTTCCTCTTCTCATTGTTTTTGGTGATTTATGTCTTKIQIFLFSLFLVIYVLTLLGNGACCTTGCTGGGAAATGGAGCCATCATCTATGCAGTGAGATGCAACCCACTACTACAGAIIYAVRCNPLLHYPMYFLLCACCCCCATGTACTTTCTGCTGGGAAATTTTGCCTTCCTTGAGATCTGGTATGTGCCGNFAFLEIWYVPSTIPNMLVNCTCCACTATTCCTAACATGCTAGTCAACATTCTCTCCAAGACCAAGGCCATCTCATTILSKTKAISFSGCFLQFYFFFTTCTGGGTGCTTCCTCCAGTTCTATTTCTTCTTTTCACTGGGAACAACTGAATGTCTSLGTTECLFLAVMAYDRYLAICTTTCTGGCAGTAATGGCTTATGATCGATACCTGGCCATCTGCCACCCACTGCAGTACHPLQYPAIMTVRFCGKLVSFCCCTGCCATCATGACTGTAAGGTTCTGTGGTAAGCTGGTGTCTTTCTGTTGGCTTATCWLIGFLGYPIPIFYISQLPFTGGATTCCTTGGATACCCAATTCCCATTTTCTACATCTCCCAACTCCCCTTCTGTGGCGPNIIDHFLCDMDPLMALSCTCCTAATATCATTGATCACTTCCTGTGTGACATGGACCCATTGATGGCTCTATCCTGAPAPITECIFYTQSSLVLFFTTGCCCCAGCTCCCATAACTGAATGTATTTTCTATACTCAGAGCTCCCTTGTCCTCTTSMYILRSYILLLTAVFQVPSATTTCACTAGTATGTACATTCTTCGATCCTATATCCTGTTACTAACAGCTGTTTTTCAAGRRKAFSTCGSHLVVVSLFYGGTCCCTTCTGCAGCTGGTCGGAGAAAAGCCTTCTCTACCTGTGGTTCTCATTTGGTGTVMVMYVSPTYGIPTLLQKITGTGGTATCTCTTTTCTATGGGACAGTCATGGTAATGTATGTAAGTCCTACATATGGLTLVYSVTTPLFNPLIYTLRNGATCCCAACTTTATTGCAGAAGATCCTCACACTGGTATATTCAGTAACGACTCCTCTKDMKLALRNVLFGMRIRQNSTTTTAATCCTCTGATCTATACTCTTCGTAATAAGGACATGAAACTCGCTCTGAGAAATGTCCTGTTTGGAATGAGAATTCGTCAAAATTCGTGAGCCAAAGATGMAC024399_A135/136ACAATGGATGTGGGCAATAAGTCTACCATGTCTGAATTTGTTTTGCTGGGGCTCTCTMDVGNKSTMSEFVLLGLSNSWAATTCCTGGGAACTACAGATGTTTTTCTTTATGGTGTTTTCATTGCTTTATGTGGCAELQMFFFMVFSLLYVATMVGNACAATGGTGGGTAACAGCCTCATAGTCATCACAGTTATAGTGGACCCTCACCTACACSLIVITVIVDPHLHSPMYFLLTCTCCTATGTATTTCCTGCTTACCAATCTTTCAATCATTGATATGTCTCTTGCTTCTTNLSIIDMSLASFATPKMITDTTCGCCACCCCAAAGATGATTACAGATTACCTAACAGGTCACAAAACCATCTCTTTTYLTGHKTISFDGCLTQIFFLHGATGGCTGCCTTACCCAGATATTCTTTCTCCACCTTTTCACTGGAACTGAGATCATCLFTGTEIILLMAMSFDRYIAITTACTCATGGCCATGTCCTTTGATAGGTATATTGCAATATGCAAGCCCCTGCACTATCKPLHYASVISPQVCVALVVAGCTTCTGTCATTAGTCCCCAGGTGTGTGTTGCTCTCGTGGTGGCTTCCTGGATTATGSWIMGVMHSMSQVIFALTLPFGGAGTTATGCATTCAATGAGTCAGGTCATATTTGCCCTCACGTTACCATTCTGTGGTCGPYEVDSFFCDLPVVFQLACCCCTATGAGGTAGACAGCTTTTTCTGTGACCTTCCTGTGGTGTTCCAGTTGGCTTGTVDTYVLGLFMISTSGIIALSCGTGGATACTTATGTTCTGGGCCTCTTTATGATCTCAACAAGTGGCATAATTGCGTTGFIVLFNSYVIVLVTVKHHSSRTCCTGTTTTATTGTTTTATTTAATTCATATGTTATTGTCCTGGTTACTGTGAAGCATGSSKALSTCTAHFIVVFLFFGCATTCTTCCAGAGGATCATCTAAGGCCCTTTCTACTTGTACAGCTCATTTCATTGTTPCIFIYMWPLSSFLTDKILSVGTCTTCTTGTTCTTTGGGCCATGCATCTTCATCTACATGTGGCCACTAAGCAGCTTTFYTIFTPTLNPIIYTLRNQEVCTCACAGACAAGATTCTGTCTGTGTTTTATACCATCTTTACTCCCACTCTGAACCCAKIAMRKLKNRFLNFNKAMPSATAATCTATACTTTGAGGAATCAAGAAGTAAAGATAGCCATGAGGAAACTGAAAAATAGGTTTCTAAATTTTAATAAGGCAATGCCTTCATAGTTTTTSG122737711_A—137/138ATGCGAGGTTTCAACAAAACCACTGTGGTTACACAGTTCATCCTGGTGGGTTTCTCCMRGFNKTTVVTQFILVGFSSLAGCCTGGGGGAGCTCCAGCTGCTGCTTTTTGTCATCTTTCTTCTCCTATACTTGACAGELQLLLFVIFLLLYLTILVAATCCTGGTGGCCAATGTGACCATCATGGCCGTTATTCGCTTCAGCTGGACTCTCCACNVTIMAVIRFSWTLHTPMYGFACTCCCATGTATGGCTTTCTATTCATCCTTTCATTTTCTGAGTCCTGCTACACTTTTLFILSFSESCYTFVIIPQLLVGTCATCATCCCTCAGCTGCTGGTCCACCTGCTCTCAGACACCAAGACCATCTCCTTCHLLSDTKTISFMACATQLFFFATGGCCTGTGCCACCCAGCTGTTCTTTTTCCTTGGCTTTGCTTGCACCAACTGCCTCLGFACTNCLLIAVMGYDRYVACTCATTGCTGTGATGGGATATGATCGCTATGTAGCAATTTGTCACCCTCTGAGGTACICHPLRYTLIINKRLGLELISACACTCATCATAAACAAAAGGCTGGGGTTGGAGTTGATTTCTCTCTCAGGAGCCACALSGATGFFIALVATNLICDMRGGTTTCTTTATTGCTTTGGTGGCCACCAACCTCATTTGTGACATGCGTTTTTGTGGCFCGPNRVNHYFCDMAPVIKLACCCAACAGGGTTAACCACTATTTCTGTGACATGGCACCTGTTATCAAGTTAGCCTGCCTDTHVKELALFSLSILVIMVACTGACACCCATGTGAAAGAGCTGGCTTTATTTAGCCTCAGCATCCTGGTAATTATGPFLLILISYGFIVNTILKIPSGTGCCTTTTCTGTTAATTCTCATATCCTATGGCTTCATAGTTAACACCATCCTGAAGAEGKKAFVTCASHLTVVFVHYATCCCCTCAGCTGAGGGCAAGAAGGCCTTTGTCACCTGTGCCTCACATCTCACTGTGGCASIIYLRPKSKSASDKDQLGTCTTTGTCCACTATGGCTGTGCCTCTATCATCTATCTGCGGCCCAAGTCCAAGTCTVAVTYTVVTPLLNPLVYSLRNGCCTCAGACAAGGATCAGTTGGTGGCAGTGACCTACACAGTGGTTACTCCCTTACTTKEVKTALKRVLGMPVATKMSAATCCTCTTGTCTACAGTCTGAGGAACAAAGAGGTAAAAACTGCATTGAAAAGAGTTCTTGGAATGCCTGTGGCAACCAAGATGAGCTAACAAAAAATAATAATAAAATTAACTAGCG50149-02139/140CTTGGCTTCTATGACATCCCTGAACTGCATTTCTTGTTTTTTATTGTATTCACTGCTLGFYDIPELHFLFFIVFTAVYGTCTATGTCTTCATCATCATAGGGAATATGCTGATTATTGTAGCAGTGGTTAGCTCCVFIIIGNMLIIVAVVSSQRLHCAGAGGCTCCACAAACCCATGTATATTTTCTTGGCGAATCTGTCCTTCCTGGATATTKPMYIFLANLSFLDILYTSAVCTCTACACCTCCGCAGTGATGCCAAAAATGCTGGAGGGCTTCCTGCAAGAAGCAACTMPKMLEGFLQEATISVAGCLLATCTCTGTGGCTGGTTGCTTGCTCCAGTTCTTTATCTTCGGCTCTCTAGCCACAGCTQFFIFGSLATAECLLLAVMAYGAATGCTTACTGCTGGCTGTCATGGCATATGACCGCTACCTGGCAATTTGCTACCCADRYLAICYPLHYPLLMGPRRYCTCCACTACCCACTCCTGATGGGGCCCAGACGGTACATGGGGCTGGTGGTCACAACCMGLVVTTWLSGFVVDGLVVALTGGCTCTCTGGATTTGTGGTAGATGGACTGGTTGTGGCCCTGGTGGCCCAGCTGAGGVAQLRFCGPNHIDQFYCDFMLTTCTGTGGCCCCAACCACATTGACCAGTTTTACTGTGACTTTATGCTTTTCGTGGGCFVGLACSDPRVAQVTTLILSVCTGGCTTGCTCGGATCCCAGAGTGGCTCAGGTGACAACTCTCATTCTGTCTGTGTTCFCLTIPFGLILTSYARIVVAVTGCCTCACTATTCCTTTTGGACTGATTCTGACATCTTATGCCAGAATTGTGGTGGCALRVPAGASRRRAFSTCSSHLAGTGCTGAGAGTTCCTGCTGGGGCAAGCAGGAGAAGGGCTTTCTCCACATGCTCCTCCVVTTFYGTLMIFYVAPSAVHSCACCTAGCTGTAGTGACCACATTCTATGGAACGCTCATGATCTTTTATGTTGCACCCQLLSKVFSLLYTVVTPLFNPVTCTGCTGTCCATTCCCAGCTCCTCTCCAAGGTCTTCTCCCTGCTCTACACTGTGGTCIYTMRNKEVHQALRKILCIKQACCCCTCTCTTCAATCCTGTGATCTATACCATGAGGAACAAGGAGGTGCATCAGGCATETLDCTTCGGAAGATTCTCTGTATCAAACAAACTGAAACACTTGATTGAAGGAGAGGMAL365336_A141/142GTGACGGGGATGATCAACAGCAGTGTCAGTAGTGACTTCATTCTGGTGGGTTTCTCAMINSSVSSDFILVGFSDQPQLGATCAGCCTCAGTTGGAAAGGAGACTCTTCATTGTAGTTTTAATTTCCTATCTTCTCERRLFIVVLISYLLTLVGNTIACTCTGGTGGGAAATACAATCATTATTTTGATTTCTTCAATAGATTCTAAACTCAAAIILISSIDSKLKTPMYFFLTHACCCCTATGTACTTTTTTCTCACTCACCTCTCCTTTGTTGACATCTGTTTCACCACCLSFVDICFTTSIVPQLLWNLKAGTATTGTCCCCCAACTGCTATGGAACCTGAAAGGACCAGCCAAGACTATCACAGCTGPAKTITAVGCAVQLYVSLTLGTGGGCTGTGCAGTGCAGCTTTATGTCTCTCTGACTCTGGGCTCTACTGAATGTATTGSTECILLAVMAFDRYAAVCKCTCCTGGCAGTAATGGCTTTCGATCGCTATGCTGCTGTCTGCAAACCTCTCCACTATPLHYVAVMNPQLCRALAGISWGTAGCAGTGATGAATCCACAGCTCTGCCGGGCTCTAGCAGGAATCTCATGGCTCAGTLSGIGNALIQGTITLWLPRCGGGAATAGGAAATGCTCTCATCCAAGGAACAATCACTCTTTGGCTTCCACGTTGTGGCHLWLHHFFCEVPSMIKLACVDCACCTGTGGCTCCACCACTTCTTCTGTGAAGTCCCCTCCATGATCAAGCTTGCCTGTIHANEVQLFVASLVLLLLPLAGTGGACATTCATGCCAATGAGGTCCAACTCTTTGTAGCCTCATTGGTCTTGCTGCTCLILTSYGHIAKAVIRIKSSQACTACCCTTAGCCCTGATACTGACGTCCTATGGACATATAGCTAAGGCAGTTATAAGAWRRALGTCGSHLMVVSLFYGSATCAAGTCATCCCAGGCTTGGCGTAGAGCCCTGGGCACATGTGGATCCCACTTGATGITAIYIQPNSSYAHTHGKFISGTTGTGTCTCTCTTTTATGGGAGCATCACAGCCATCTACATCCAGCCGAACAGTTCALFYTVMTPTLNPLIYTLRNKETATGCCCACACCCATGGGAAGTTCATCTCTCTCTTTTACACTGTTATGACCCCGACCVKHALGRLFNRASGVCTTAATCCCCTCATCTACACACTGAGGAATAAGGAGGTGAAAGGGGCTCTCGGACGGCTCTTCAATAGAGCCTCTGGAGTGTGACAGGACTTTACGATTAGAGGATCCTGAACATACTGMAL359352_A143/144GTGACGGGGATGATCAACAGCAGTGTCAGTAGTGACTTCATTCTGGTGGGTTTCTCAMINSSVSSDFILVGFSDQPQLGATCAGCCTCAGTTGGAAAGGAGACTCTTCATTGTAGTTTTAATTTCCTATCTTCTCERRLFIVVLISYLLTLVGNTIACTCTGGTGGGAAATACAATCATTATTTTGATTTCTTCAATAGATTCTAAACTCAAAIILISSIDSKLKTPMYFFLTHACCCCTATGTACTTTTTTCTCACTCACCTCTCCTTTGTTGACATCTGTTTCACCACCLSFVDICFTTSIVPQLLWNLKAGTATTGTCCCCCAACTGCTATGGAACCTGAAAGGACCAGCCAAGACTATCACAGCTGPAKTITAVGCAVQLYVSLTLGTGGGCTGTGCAGTGCAGCTTTATGTCTCTCTGACTCTGGGCTCTACTGAATGTATTGSTECILLAVMAFDRYAAVCKCTCCTGGCAGTAATGGCTTTCGATCGCTATGCTGCTGTCTGCAAACCTCTCCACTATPLHYVAVMNPQLCRALAGISWGTAGCAGTGATGAATCCACAGCTCTGCCGGGCTCTAGCAGGAATCTCATGGCTCAGTLSGIGNALIQGTITLWLPRCGGGAATAGGAAATGCTCTCATCCAAGGAACAATCACTCTTTGGCTTCCACGTTGTGGCHLWLHHFFCEVPSMIKLACVDCACCTGTGGCTCCACCACTTCTTCTGTGAAGTCCCCTCCATGATCAAGCTTGCCTGTIHANEVQLFVASLVLLLLPLAGTGGACATTCATGCCAATGAGGTCCAACTCTTTGTAGCCTCATTGGTCTTGCTGCTCLILTSYGHIAKAVIRIKSSQACTACCCTTAGCCCTGATACTGACGTCCTATGGACATATAGCTAAGGCAGTTATAAGAWRRALGTCGSHLMVVSLFYGSATCAAGTCATCCCAGGCTTGGCGTAGAGCCCTGGGCACATGTGGATCCCACTTGATGITAIYIQPNSSYAHTHGKFISGTTGTGTCTCTCTTTTATGGGAGCATCACAGCCATCTACATCCAGCCGAACAGTTCALFYTVMTPTLNPLIYTLRNKETATGCCCACACCCATGGGAAGTTCATCTCTCTCTTTTACACTGTTATGACCCCGACCVKGALGRLFNRASGVCTTAATCCCCTCATCTACACACTGAGGAATAAGGAGGTGAAAGGGGCTCTCGGACGGCTCTTCAATAGAGCCTCTGGAGTGTGACAGGACTTTACGATTAGAGGATCCTGAACATACTCG56115-02145/146AGGACTAACCAATGACTCAGAACTGCAGGTTCCCCTCTTTATAACGTTCCCCTTCATGLTNDSELQVPLFITFPFIYICTATATTATCACTCTGGTTGGAAACCTGGGAATTATTGTATTGATATTCTGGGATTCITLVGNLGIIVLIFWDSCLHNCTGTCTCCACAATCCCATGTACTTTTTTCTCAGTAACTTGTCTCTAGTGGACTTTTGPMYFFLSNLSLVDFCYSSAVTCTACTCTTCAGCTGTCACTCCCATCGTCATGGCTGGATTCCTTATAGAAGACAAGGTPIVMAGFLIEDKVISYNACAACATCTCTTACAATGCATGTGCTGCTCAAATGTATATCTTTGTAGCTTTTGCCACTGTQMYIFVAFATVENYLLASMAYGGAAAATTACCTCTTGGCCTCAATGGCCTATGACCGCTATGCAGCAGTGTGCAAACCDRYAAVCKPLHYTTTMTTTVCCCTACATTACACCACAACCATGACAACAACTGTGTGTGCTCGTCTGGCCATAGGCTCARLAIGSYLCGFLNASIHTGDCTACCTCTGTGGTTTCCTGAATGCCTCCATCCACACTGGGGACACATTTAGTCTCTCTFSLSFCKSNEVHHFFCDIPATTTCTGTAAGTCCAATGAAGTCCATCACTTTTTCTGTGATATTCCAGCAGTCATGGTVMVLSCSDRHISELVLIYVVSTCTCTCTTGCTCTGATAGACATATTAGCGAGCTTGTTCTTATTTATGTTGTGAGCTTFNIFIALLVILISYTFIFITICAATATCTTTATAGCTCTCCTGGTTATCTTGATATCCTACACATTCATTTTTATCACLKMHSASVYQKPLSTCASHFICATCCTAAAGATGCACTCAGCTTCAGTATACCAGAAGCCTTTGTCCACCTGTGCCTCAVGIFYGTIIFMYLQPSSSHSTCATTTCATTGCAGTCGGCATCTTCTATGGGACTATTATCTTCATGTACTTACAACCMDTDKMAPVFYTMVIPMLNPLCAGCTCCAGTCACTCCATGGACACAGACAAAATGGCACCTGTATTCTATACAATGGTVYSLRNDEVDSAFKKVVEKAKCATCCCCATGCTGAACCCTCTGGTCTATAGTCTGAGGAACAAGGAAGTGAAGAGTGCLSVGWSVATTCAAGAAAGTTGTTGAGAAGGCAAAATTGTCTGTAGGATGGTCAGTTTAACATTCG55993-02147/148CCCCTTGTCTCCTCACACAATGACCCTGGGATCCCTGGGAAACAGCAGCAGCAGCGTMTLGSLGNSSSSVSATFLLSGTTCTGCTACCTTCCTGCTGAGTGGCATCCCTGGGCTGGAGCGCATGCACATCTGGATIPGLERMHIWISIPLCFMYLVCTCCATCCCACTGTGCTTCATGTATCTGGTCTCCATCCCGGGCAACTGCACAATTCTSIPGNCTILFIIKTERSLHEPTTTTATCATTAAAACAGAGCGCTCACTTCATGAACCTATGTATCTCTTCCTGTCCATMYLFLSMLALIDLGLSLCTLPGCTGGCTCTGATTGACCTGGGTCTCTCCCTTTGCACTCTCCCTACAGTCCTGGGCATTVLGIFWVGEREISHDACFAQCTTTTGGGTTGGAGAACGAGAAATTAGCCATGATGCCTGCTTTGCTCAGCTCTTTTTLFFIHCFSFLESSVLLSMAFDCATTCACTGCTTCTCCTTCCTCGAGTCCTCTGTGCTACTGTCTATGGCCTTTGACCGRFVAICHPLHYVSILTNTVIGCTTTGTGGCTATCTGCCACCCCTTGCACTATGTTTCCATTCTCACCAACACAGTCATRIGLVSLGRSVALIFPLPFMLTGGCAGGATTGGCCTGGTCTCTCTGGGTCGTAGTGTAGCACTCATTTTTCCATTACCKRFPYCGSPVLSHSYCLHQEVTTTTATGCTCAAAAGATTCCCCTATTGTGGCTCCCCAGTTCTCTCACATTCTTATTGMKLACADMKANSIYGMFVIVSTCTCCACCAAGAAGTGATGAAATTGGCCTGTGCCGACATGAAGGCCAACAGCATCTATVGIDSLLILFSYALILRTVLCGGCATGTTTGTCATCGTCTCTACAGTGGGTATAGACTCACTGCTCATCCTCTTCTCSIASRAERFKALNTCVSHICATTATGCTCTGATCCTGCGCACCGTGCTGTCCATCGCCTCCAGGGCTGAGAGATTCAAVLLFYTPMIGLSVIHRFGKQAGGCCCTTAACACCTGTGTTTCCCACATCTGTGCTGTGCTGCTCTTCTACACTCCCATPHLVQVVMGFMYLLFPPVMNPGATTGGCCTCTCTGTCATCCATCGCTTTGGAAAGCAGGCACCCCACCTGGTCCAGGTIVYSVKTKQIRDRVTHAFCYGGTCATGGGTTTCATGTATCTTCTCTTTCCTCCTGTGATGAATCCCATTGTCTACAGTGTGAAGACCAAACAGATCCGGGATCGAGTGACGCATGCCTTTTGTTACTAACTGM445n18_A149/150AAATGGAAAACCAATGATGGGAGAAGCAAGGAACAGGACAGTAGTCCAGGAATTTATMMGEARNRTVVQEFILEGFPACCTGGAGGGATTTCCTGCTGTCCAGCATCTGGGGAATGTCCTTTTCCTGGTGCACCTVQHLGNVLFLVHLLAYLASIMGCTGGCATACCTGGCCTCCATCATGGCAAACATGCTCATAATCACCATCACCTGGGCANMLIITITWADHHLQTPMYFTGACCATCACCTCCAGACACCTATGTATTTCTTCCTCAACAGTTTTTCCTTCTGTGAFLNSFSFCECCFITTVIPKLLATGCTGTTTTATCACCACAGTTATTCCTAAACTTCTGGTCATCTTTCTTTCAGGCAGVIFLSGRQIIPFTTCLMQSFSGCAAATAATCCCCTTTACTACTTGTCTCATGCAGTCCTTTTCATTTTTATTTCTTGGFLFLGSTVFFLMAVMSLDRYLGTCAACAGTTTTCTTCCTTATGGCTGTGATGTCCTTGGATAGATACCTGGCCATTTGAICKPLHYSTIMSLRTSFHLVCAAGCCTCTGCATTACTCCACCATCATGAGCCTGAGGACTAGCTTCCACCTGGTCACTVCFVVGFTLITGLMVKVSQLTGTCTGCTTTGTCGTGGGCTTCACTCTCATCACTGGTCTCATGGTGAAGGTTTCCCASFCGPHVIPHFFRDLGPLIQLGTTATCTTTCTGTGGACCCCATGTCATCCCTCACTTCTTCCGTGACCTCGGCCCTCTSCSDTRSTETLAFVLVSFVLFGATCCAACTCTCCTGTTCTGACACCAGATCTACTGAAACGTTGGCCTTTGTCCTTGTTSLIITIIAYGNIVVTIVRLPTTCATTCGTTCTTTTTACATCCCTCATTATAACCATCATTGCATATGGCAACATAGTSAKERQKAFSTCSSHLIVLSLAGTCACAATTGTACGACTCCCATCAGCCAAGGAGCGGCAGAAAGCTTTCTCCACCTGVYGSCVFIYVKPKQMDRLDSNCTCCTCTCACCTCATTGTCCTCTCTCTGGTGTATGGCAGCTGTGTCTTCATATATGTRMAALVNTVVTPLLNPIIYTLGAAGCCGAAGCAAATGGACAGGCTGGACTCCAACAGAATGGCTGCTCTTGTGAACACRNKQVHQALRDAQSRMKLAGTGGTGACCCCACTGCTGAACCCGATCATTTACACTCTGCGGAACAAGCAGGTCCACCAGGCTCTGAGGGATGCTCAGTCCAGAATGAAATTGTAAAAACAGAATCACAACCTCCCACG90341_01151/152CGGTTGCCCCTGCTGAATTCGTCCTCCTGGGCATCACAAATCGCTGGGACCTGCGTGVAPAEFVLLGITNRWDLRVALTGGCCCTCCTCCTGACCTGCCTGCCTGTCTACCTGGTGAGCCTGCTGGGAAACATGGLLTCLPVYLVSLLGNMGMALLGCATGGCGCTGCTGATCCGCATGGATGCCCGGCTCCACACACCTATGTACTTCTTCCIRMDARLHTPMYFFLANLSLLTGGCCAACCTCTCCCTGCTGGATGCCTGCTATTCCTCCGCCATCGGCCCCAAGATGCDACYSSAIGPKMLVDLLLPRATAGTGGACCTGCTGCTGCCCCGAGCCACCATCCCTTACACAGCCTGTGCCCTCCAGATIPYTACALQMFVFAGLADTETGTTTGTCTTTGCAGGTCTGGCTGATACTGAGTGTTGCTTGCTGGCAGCCATGGCCTCCLLAAMAYDRYVAIRNPLLYATGACCGCTACGTGGCCATCAGAAACCCACTTCTCTATACAACAGCTATGTCGCAGCTTAMSQRLCLALLGASGLGGAGTCTATGCCTGGCCTTGCTGGGAGCATCAGGCCTGGGTGGGGCAGTGAGTGCCTTTGVSAFVHTTLTFRLSFCRSRKITTCACACAACCCTCACCTTCCGTCTGAGCTTCTGCCGCTCCCGGAAGATCAATAGCTNSFFCDIPPLLAISCSDTSLNTCTTCTGCGATATCCCTCCACTGCTGGCCATCTCGTGCAGTGACACCAGTCTCAATGELLLFAICGFIQTATVLAITVAACTCCTTCTCTTCGCCATCTGTGGCTTCATCCAGACAGCCACGGTGTTAGCTATCASYGFIAGAVIHMRSVEGSRRACAGTGTCTTATGGCTTCATCGCTGGGGCTGTGATCCACATGCGCTCGGTCGAGGGCAASTGGSHLTAVAMMYGTLIFMGTCGGCGAGCAGCCTCCACCGGTGGTTCCCACCTCACAGCCGTGGCCATGATGTACGYLRPSSSYALDTDKMASVFYTGGACACTCATTTTCATGTACCTGCGCCCCAGCTCCAGCTATGCCCTGGACACTGACALVIPSLNPLIYSLRNKEVKEAAGATGGCCTCTGTGTTCTATACCCTGGTCATCCCGTCTCTCAACCCACTCATCTACALRQTWSRFHCPGQGSQGCCTCCGCAATAAGGAGGTCAAGGAGGCCCTCAGGCAGACCTGGAGCCGATTCCACTGTCCAGGGCAGGGGTCCCAGTGATTGGTCCAGGGAGGCTGGGTAGGTCTGACTATGAGGGGATGAGGAAG

[0037] GPCRX Nucleic Acids and Polypeptides

[0038] One aspect of the invention pertains to isolated nucleic acid molecules that encode GPCRX polypeptides or biologically active portions thereof. Also included in the invention are nucleic acid fragments sufficient for use as hybridization probes to identify GPCRX-encoding nucleic acids (e.g., GPCRX mRNAs) and fragments for use as PCR primers for the amplification and/or mutation of GPCRX nucleic acid molecules. As used herein, the term “nucleic acid molecule” is intended to include DNA molecules (e.g., cDNA or genomic DNA), RNA molecules (e.g., mRNA), analogs of the DNA or RNA generated using nucleotide analogs, and derivatives, fragments and homologs thereof. The nucleic acid molecule may be single-stranded or double-stranded, but preferably is comprised double-stranded DNA.

[0039] An GPCRX nucleic acid can encode a mature GPCRX polypeptide. As used herein, a “mature” form of a polypeptide or protein disclosed in the present invention is the product of a naturally occurring polypeptide or precursor form or proprotein. The naturally occurring polypeptide, precursor or proprotein includes, by way of nonlimiting example, the full-length gene product, encoded by the corresponding gene. Alternatively, it may be defined as the polypeptide, precursor or proprotein encoded by an ORF described herein. The product “mature” form arises, again by way of nonlimiting example, as a result of one or more naturally occurring processing steps as they may take place within the cell, or host cell, in which the gene product arises. Examples of such processing steps leading to a “mature” form of a polypeptide or protein include the cleavage of the N-terminal methionine residue encoded by the initiation codon of an ORF, or the proteolytic cleavage of a signal peptide or leader sequence. Thus a mature form arising from a precursor polypeptide or protein that has residues 1 to N, where residue 1 is the N-terminal methionine, would have residues 2 through N remaining after removal of the N-terminal methionine. Alternatively, a mature form arising from a precursor polypeptide or protein having residues 1 to N, in which an N-terminal signal sequence from residue 1 to residue M is cleaved, would have the residues from residue M+1 to residue N remaining. Further as used herein, a “mature” form of a polypeptide or protein may arise from a step of post-translational modification other than a proteolytic cleavage event. Such additional processes include, by way of non-limiting example, glycosylation, myristoylation or phosphorylation. In general, a mature polypeptide or protein may result from the operation of only one of these processes, or a combination of any of them.

[0040] The term “probes”, as utilized herein, refers to nucleic acid sequences of variable length, preferably between at least about 10 nucleotides (nt), 100 nt, or as many as approximately, e.g., 6,000 nt, depending upon the specific use. Probes are used in the detection of identical, similar, or complementary nucleic acid sequences. Longer length probes are generally obtained from a natural or recombinant source, are highly specific, and much slower to hybridize than shorter-length oligomer probes. Probes may be single- or double-stranded and designed to have specificity in PCR, membrane-based hybridization technologies, or ELISA-like technologies.

[0041] The term “isolated” nucleic acid molecule, as utilized herein, is one, which is separated from other nucleic acid molecules which are present in the natural source of the nucleic acid. Preferably, an “isolated” nucleic acid is free of sequences which naturally flank the nucleic acid (i.e., sequences located at the 5′- and 3′-termini of the nucleic acid) in the genomic DNA of the organism from which the nucleic acid is derived. For example, in various embodiments, the isolated GPCRX nucleic acid molecules can contain less than about 5 kb, 4 kb, 3 kb, 2 kb, 1 kb, 0.5 kb or 0.1 kb of nucleotide sequences which naturally flank the nucleic acid molecule in genomic DNA of the cell/tissue from which the nucleic acid is derived (e.g. brain, heart, liver, spleen, etc.). Moreover, an “isolated” nucleic acid molecule, such as a cDNA molecule, can be substantially free of other cellular material or culture medium when produced by recombinant techniques, or of chemical precursors or other chemicals when chemically synthesized.

[0042] A nucleic acid molecule of the invention, e.g., a nucleic acid molecule having the nucleotide sequence of SEQ ID NOS:1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, 75, 77, 79, 81, 83, 85, 87, 89, 91, 93, 95, 97, 99, 101, 103, 105, 107, 109, 111, 113, 115, 117, 119, 121, 123, 125, 127, 129, 131, 133, 135, 137, 139, 141, 143, 145, 147, 149 and 151, or a complement of this aforementioned nucleotide sequence, can be isolated using standard molecular biology techniques and the sequence information provided herein. Using all or a portion of the nucleic acid sequence of SEQ ID NOS:1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, 75, 77, 79, 81, 83, 85, 87, 89, 91, 93, 95, 97, 99, 101, 103, 105, 107, 109, 111, 113, 115, 117, 119, 121, 123, 125, 127, 129, 131, 133, 135, 137, 139, 141, 143, 145, 147, 149 and 151 as a hybridization probe, GPCRX molecules can be isolated using standard hybridization and cloning techniques (e.g., as described in Sambrook, et al., (eds.), MOLECULAR CLONING: A LABORATORY MANUAL 2nd Ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., 1989; and Ausubel, et al., (eds.), CURRENT PROTOCOLS IN MOLECULAR BIOLOGY, John Wiley & Sons, New York, N.Y., 1993.)

[0043] A nucleic acid of the invention can be amplified using cDNA, mRNA or alternatively, genomic DNA, as a template and appropriate oligonucleotide primers according to standard PCR amplification techniques. The nucleic acid so amplified can be cloned into an appropriate vector and characterized by DNA sequence analysis. Furthermore, oligonucleotides corresponding to GPCRX nucleotide sequences can be prepared by standard synthetic techniques, e.g., using an automated DNA synthesizer.

[0044] As used herein, the term “oligonucleotide” refers to a series of linked nucleotide residues, which oligonucleotide has a sufficient number of nucleotide bases to be used in a PCR reaction. A short oligonucleotide sequence may be based on, or designed from, a genomic or cDNA sequence and is used to amplify, confirm, or reveal the presence of an identical, similar or complementary DNA or RNA in a particular cell or tissue. Oligonucleotides comprise portions of a nucleic acid sequence having about 10 nt, 50 nt, or 100 nt in length, preferably about 15 nt to 30 nt in length. In one embodiment of the invention, an oligonucleotide comprising a nucleic acid molecule less than 100 nt in length would further comprise at least 6 contiguous nucleotides of SEQ ID NOS:1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, 75, 77, 79, 81, 83, 85, 87, 89, 91, 93, 95, 97, 99, 101, 103, 105, 107, 109, 111, 113, 115, 117, 119, 121, 123, 125, 127, 129, 131, 133, 135, 137, 139, 141, 143, 145, 147, 149 and 151, or a complement thereof. Oligonucleotides may be chemically synthesized and may also be used as probes.

[0045] In another embodiment, an isolated nucleic acid molecule of the invention comprises a nucleic acid molecule that is a complement of the nucleotide sequence shown in SEQ ID NOS:1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, 75, 77, 79, 81, 83, 85, 87, 89, 91, 93, 95, 97, 99, 101, 103, 105, 107, 109, 111, 113, 115, 117, 119, 121, 123, 125, 127, 129, 131, 133, 135, 137, 139, 141, 143, 145, 147, 149 and 151, or a portion of this nucleotide sequence (e.g., a fragment that can be used as a probe or primer or a fragment encoding a biologically-active portion of an GPCRX polypeptide). A nucleic acid molecule that is complementary to the nucleotide sequence shown in SEQ ID NOS:1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, 75, 77, 79, 81, 83, 85, 87, 89, 91, 93, 95, 97, 99, 101, 103, 105, 107, 109, 111, 113, 115, 117, 119, 121, 123, 125, 127, 129, 131, 133, 135, 137, 139, 141, 143, 145, 147, 149 and 151 is one that is sufficiently complementary to the nucleotide sequence shown in SEQ ID NOS:1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, 75, 77, 79, 81, 83, 85, 87, 89, 91, 93, 95, 97, 99, 101, 103, 105, 107, 109, 111, 113, 115, 117, 119, 121, 123, 125, 127, 129, 131, 133, 135, 137, 139, 141, 143, 145, 147, 149 and 151 that it can hydrogen bond with little or no mismatches to the nucleotide sequence shown SEQ ID NOS:1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, 75, 77, 79, 81, 83, 85, 87, 89, 91, 93, 95, 97, 99, 101, 103, 105, 107, 109, 111, 113, 115, 117, 119, 121, 123, 125, 127, 129, 131, 133, 135, 137, 139, 141, 143, 145, 147, 149 and 151, thereby forming a stable duplex.

[0046] As used herein, the term “complementary” refers to Watson-Crick or Hoogsteen base pairing between nucleotides units of a nucleic acid molecule, and the term “binding” means the physical or chemical interaction between two polypeptides or compounds or associated polypeptides or compounds or combinations thereof. Binding includes ionic, non-ionic, van der Waals, hydrophobic interactions, and the like. A physical interaction can be either direct or indirect. Indirect interactions may be through or due to the effects of another polypeptide or compound. Direct binding refers to interactions that do not take place through, or due to, the effect of another polypeptide or compound, but instead are without other substantial chemical -intermediates.

[0047] Fragments provided herein are defined as sequences of at least 6 (contiguous) nucleic acids or at least 4 (contiguous) amino acids, a length sufficient to allow for specific hybridization in the case of nucleic acids or for specific recognition of an epitope in the case of amino acids, respectively, and are at most some portion less than a full length sequence. Fragments may be derived from any contiguous portion of a nucleic acid or amino acid sequence of choice. Derivatives are nucleic acid sequences or amino acid sequences formed from the native compounds either directly or by modification or partial substitution. Analogs are nucleic acid sequences or amino acid sequences that have a structure similar to, but not identical to, the native compound but differs from it in respect to certain components or side chains. Analogs may be synthetic or from a different evolutionary origin and may have a similar or opposite metabolic activity compared to wild type. Homologs are nucleic acid sequences or amino acid sequences of a particular gene that are derived from different species.

[0048] Derivatives and analogs may be full length or other than full length, if the derivative or analog contains a modified nucleic acid or amino acid, as described below. Derivatives or analogs of the nucleic acids or proteins of the invention include, but are not limited to, molecules comprising regions that are substantially homologous to the nucleic acids or proteins of the invention, in various embodiments, by at least about 70%, 80%, or 95% identity (with a preferred identity of 80-95%) over a nucleic acid or amino acid sequence of identical size or when compared to an aligned sequence in which the alignment is done by a computer homology program known in the art, or whose encoding nucleic acid is capable of hybridizing to the complement of a sequence encoding the aforementioned proteins under stringent, moderately stringent, or low stringent conditions. See e.g. Ausubel, et al., CURRENT PROTOCOLS IN MOLECULAR BIOLOGY, John Wiley & Sons, New York, N.Y., 1993, and below.

[0049] A “homologous nucleic acid sequence” or “homologous amino acid sequence,” or variations thereof, refer to sequences characterized by a homology at the nucleotide level or amino acid level as discussed above. Homologous nucleotide sequences encode those sequences coding for isoforms of GPCRX polypeptides. Isoforms can be expressed in different tissues of the same organism as a result of, for example, alternative splicing of RNA. Alternatively, isoforms can be encoded by different genes. In the invention, homologous nucleotide sequences include nucleotide sequences encoding for an GPCRX polypeptide of species other than humans, including, but not limited to: vertebrates, and thus can include, e.g., frog, mouse, rat, rabbit, dog, cat cow, horse, and other organisms. Homologous nucleotide sequences also include, but are not limited to, naturally occurring allelic variations and mutations of the nucleotide sequences set forth herein. A homologous nucleotide sequence does not, however, include the exact nucleotide sequence encoding human GPCRX protein. Homologous nucleic acid sequences include those nucleic acid sequences that encode conservative amino acid substitutions (see below) in SEQ ID NOS:1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, 75, 77, 79, 81, 83, 85, 87, 89, 91, 93, 95, 97, 99, 101, 103, 105, 107, 109, 111, 113, 115, 117, 119, 121, 123, 125, 127, 129, 131, 133, 135, 137, 139, 141, 143, 145, 147, 149 and 151, as well as a polypeptide possessing GPCRX biological activity. Various biological activities of the GPCRX proteins are described below.

[0050] An GPCRX polypeptide is encoded by the open reading frame (“ORF”) of an GPCRX nucleic acid. An ORF corresponds to a nucleotide sequence that could potentially be translated into a polypeptide. A stretch of nucleic acids comprising an ORF is uninterrupted by a stop codon. An ORF that represents the coding sequence for a full protein begins with an ATG “start” codon and terminates with one of the three “stop” codons, namely, TAA, TAG, or TGA. For the purposes of this invention, an ORF may be any part of a coding sequence, with or without a start codon, a stop codon, or both. For an ORF to be considered as a good candidate for coding for a bonafide cellular protein, a minimum size requirement is often set, e.g., a stretch of DNA that would encode a protein of 50 amino acids or more.

[0051] The nucleotide sequences determined from the cloning of the human GPCRX genes allows for the generation of probes and primers designed for use in identifying and/or cloning GPCRX homologues in other cell types, e.g. from other tissues, as well as GPCRX homologues from other vertebrates. The probe/primer typically comprises substantially purified oligonucleotide. The oligonucleotide typically comprises a region of nucleotide sequence that hybridizes under stringent conditions to at least about 12, 25, 50, 100, 150, 200, 250, 300, 350 or 400 consecutive sense strand nucleotide sequence of SEQ ID NOS:1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, 75, 77, 79, 81, 83, 85, 87, 89, 91, 93, 95, 97, 99, 101, 103, 105, 107, 109, 111, 113, 115, 117, 119, 121, 123, 125, 127, 129, 131, 133, 135, 137, 139, 141, 143, 145, 147, 149 and 151; or an anti-sense strand nucleotide sequence of SEQ ID NOS: 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, 75, 77, 79, 81, 83, 85, 87, 89, 91, 93, 95, 97, 99, 101, 103, 105, 107, 109, 111, 113, 115, 117, 119, 121, 123, 125, 127, 129, 131, 133, 135, 137, 139, 141, 143, 145, 147, 149 and 151; or of a naturally occurring mutant of SEQ ID NOS:1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, 75, 77, 79, 81, 83, 85, 87, 89, 91, 93, 95, 97, 99, 101, 103, 105, 107, 109, 111, 113, 115, 117, 119, 121, 123, 125, 127, 129, 131, 133, 135, 137, 139, 141, 143, 145, 147, 149 and 151.

[0052] Probes based on the human GPCRX nucleotide sequences can be used to detect transcripts or genomic sequences encoding the same or homologous proteins. In various embodiments, the probe further comprises a label group attached thereto, e.g. the label group can be a radioisotope, a fluorescent compound, an enzyme, or an enzyme co-factor. Such probes can be used as a part of a diagnostic test kit for identifying cells or tissues which mis-express an GPCRX protein, such as by measuring a level of an GPCRX-encoding nucleic acid in a sample of cells from a subject e.g., detecting GPCRX mRNA levels or determining whether a genomic GPCRX gene has been mutated or deleted.

[0053] “A polypeptide having a biologically-active portion of an GPCRX polypeptide” refers to polypeptides exhibiting activity similar, but not necessarily identical to, an activity of a polypeptide of the invention, including mature forms, as measured in a particular biological assay, with or without dose dependency. A nucleic acid fragment encoding a “biologically-active portion of GPCRX” can be prepared by isolating a portion SEQ ID NOS:1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, 75, 77, 79, 81, 83, 85, 87, 89, 91, 93, 95, 97, 99, 101, 103, 105, 107, 109, 111, 113, 115, 117, 119, 121, 123, 125, 127, 129, 131, 133, 135, 137, 139, 141, 143, 145, 147, 149 and 151 that encodes a polypeptide having an GPCRX biological activity (the biological activities of the GPCRX proteins are described below), expressing the encoded portion of GPCRX protein (e.g., by recombinant expression in vitro) and assessing the activity of the encoded portion of GPCRX.

[0054] GPCRX Nucleic Acid and Polypeptide Variants

[0055] The invention further encompasses nucleic acid molecules that differ from the nucleotide sequences shown SEQ ID NOS:1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, 75, 77, 79, 81, 83, 85, 87, 89, 91, 93, 95, 97, 99, 101, 103, 105, 107, 109, 111, 113, 115, 117, 119, 121, 123, 125, 127, 129, 131, 133, 135, 137, 139, 141, 143, 145, 147, 149 and 151 due to degeneracy of the genetic code and thus encode the same GPCRX proteins as that encoded by the nucleotide sequences shown in SEQ ID NOS:1, 3, 5,7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, 75, 77, 79, 81, 83, 85, 87, 89, 91, 93, 95, 97, 99, 101, 103, 105, 107, 109, 111, 113, 115, 117, 119, 121, 123, 125, 127, 129, 131, 133, 135, 137, 139, 141, 143, 145, 147, 149 and 151. In another embodiment, an isolated nucleic acid molecule of the invention has a nucleotide sequence encoding a protein having an amino acid sequence shown in SEQ ID NOS: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, 78, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 100, 102, 104, 106, 108, 110, 112, 114, 116, 118, 120, 122, 124, 126, 128, 130, 132, 134, 136, 138, 140, 142, 144, 146, 148, 150 and 152.

[0056] In addition to the human GPCRX nucleotide sequences shown in SEQ ID NOS:1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, 75, 77, 79, 81, 83, 85, 87, 89, 91, 93, 95, 97, 99, 101, 103, 105, 107, 109, 111, 113, 115, 117, 119, 121, 123, 125, 127, 129, 131, 133, 135, 137, 139, 141, 143, 145, 147, 149 and 151 it will be appreciated by those skilled in the art that DNA sequence polymorphisms that lead to changes in the amino acid sequences of the GPCRX polypeptides may exist within a population (e.g., the human population). Such genetic polymorphism in the GPCRX genes may exist among individuals within a population due to natural allelic variation. As used herein, the terms “gene” and “recombinant gene” refer to nucleic acid molecules comprising an open reading frame (ORF) encoding an GPCRX protein, preferably a vertebrate GPCRX protein. Such natural allelic variations can typically result in 1-5% variance in the nucleotide sequence of the GPCRX genes. Any and all such nucleotide variations and resulting amino acid polymorphisms in the GPCRX polypeptides, which are the result of natural allelic variation and that do not alter the functional activity of the GPCRX polypeptides, are intended to be within the scope of the invention.

[0057] Moreover, nucleic acid molecules encoding GPCRX proteins from other species, and thus that have a nucleotide sequence that differs from the human sequence SEQ ID NOS:1, 3, 5, 7,9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, 75, 77, 79, 81, 83, 85, 87, 89, 91, 93, 95, 97, 99, 101, 103, 105, 107, 109, 111, 113, 115, 117, 119, 121, 123, 125, 127, 129, 131, 133, 135, 137, 139, 141, 143, 145, 147, 149 and 151 are intended to be within the scope of the invention. Nucleic acid molecules corresponding to natural allelic variants and homologues of the GPCRX cDNAs of the invention can be isolated based on their homology to the human GPCRX nucleic acids disclosed herein using the human cDNAs, or a portion thereof, as a hybridization probe according to standard hybridization techniques under stringent hybridization conditions.

[0058] Accordingly, in another embodiment, an isolated nucleic acid molecule of the invention is at least 6 nucleotides in length and hybridizes under stringent conditions to the nucleic acid molecule comprising the nucleotide sequence of SEQ ID NOS:1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, 75, 77, 79, 81, 83, 85, 87, 89, 91, 93, 95, 97, 99, 101, 103, 105, 107, 109, 111, 113, 115, 117, 119, 121, 123, 125, 127, 129, 131, 133, 135, 137, 139, 141, 143, 145, 147, 149 and 151. In another embodiment, the nucleic acid is at least 10, 25, 50, 100, 250, 500, 750, 1000, 1500, or 2000 or more nucleotides in length. In yet another embodiment, an isolated nucleic acid molecule of the invention hybridizes to the coding region. As used herein, the term “hybridizes under stringent conditions” is intended to describe conditions for hybridization and washing under which nucleotide sequences at least 60% homologous to each other typically remain hybridized to each other.

[0059] Homologs (i.e., nucleic acids encoding GPCRX proteins derived from species other than human) or other related sequences (e.g., paralogs) can be obtained by low, moderate or high stringency hybridization with all or a portion of the particular human sequence as a probe using methods well known in the art for nucleic acid hybridization and cloning.

[0060] As used herein, the phrase “stringent hybridization conditions” refers to conditions under which a probe, primer or oligonucleotide will hybridize to its target sequence, but to no other sequences. Stringent conditions are sequence-dependent and will be different in different circumstances. Longer sequences hybridize specifically at higher temperatures than shorter sequences. Generally, stringent conditions are selected to be about 5° C. lower than the thermal melting point (Tm) for the specific sequence at a defined ionic strength and pH. The Tm is the temperature (under defined ionic strength, pH and nucleic acid concentration) at which 50% of the probes complementary to the target sequence hybridize to the target sequence at equilibrium. Since the target sequences are generally present at excess, at Tm, 50% of the probes are occupied at equilibrium. Typically, stringent conditions will be those in which the salt concentration is less than about 1.0 M sodium ion, typically about 0.01 to 1.0 M sodium ion (or other salts) at pH 7.0 to 8.3 and the temperature is at least about 30° C. for short probes, primers or oligonucleotides (e.g., 10 nt to 50 nt) and at least about 60° C. for longer probes, primers and oligonucleotides. Stringent conditions may also be achieved with the addition of destabilizing agents, such as formamide.

[0061] Stringent conditions are known to those skilled in the art and can be found in Ausubel, et al., (eds.), CURRENT PROTOCOLS IN MOLECULAR BIOLOGY, John Wiley & Sons, N.Y. (1989), 6.3.1-6.3.6. Preferably, the conditions are such that sequences at least about 65%, 70%, 75%, 85%, 90%, 95%, 98%, or 99% homologous to each other typically remain hybridized to each other. A non-limiting example of stringent hybridization conditions are hybridization in a high salt buffer comprising 6×SSC, 50 mM Tris-HCl (pH 7.5), 1 mM EDTA, 0.02% PVP, 0.02% Ficoll, 0.02% BSA, and 500 mg/ml denatured salmon sperm DNA at 65° C., followed by one or more washes in 0.2×SSC, 0.01% BSA at 50° C. An isolated nucleic acid molecule of the invention that hybridizes under stringent conditions to the sequences of SEQ ID NOS:1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, 75, 77, 79, 81, 83, 85, 87, 89, 91, 93, 95, 97, 99, 101, 103, 105, 107, 109, 111, 113, 115, 117, 119, 121, 123, 125, 127, 129, 131, 133, 135, 137, 139, 141, 143, 145, 147, 149 and 151 corresponds to a naturally-occurring nucleic acid molecule. As used herein, a “naturally-occurring” nucleic acid molecule refers to an RNA or DNA molecule having a nucleotide sequence that occurs in nature (e.g., encodes a natural protein).

[0062] In a second embodiment, a nucleic acid sequence that is hybridizable to the nucleic acid molecule comprising the nucleotide sequence of SEQ ID NOS:1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, 75, 77, 79, 81, 83, 85, 87, 89, 91, 93, 95, 97, 99, 101, 103, 105, 107, 109, 111, 113, 115, 117, 119, 121, 123, 125, 127, 129, 131, 133, 135, 137, 139, 141, 143, 145, 147, 149 and 151 or fragments, analogs or derivatives thereof, under conditions of moderate stringency is provided. A non-limiting example of moderate stringency hybridization conditions are hybridization in 6×SSC, 5× Denhardfs solution, 0.5% SDS and 100 mg/ml denatured salmon sperm DNA at 55° C., followed by one or more washes in 1×SSC, 0.1% SDS at 37° C. Other conditions of moderate stringency that may be used are well-known within the art. See, e.g., Ausubel, et al. (eds.), 1993, CURRENT PROTOCOLS IN MOLECULAR BIOLOGY, John Wiley & Sons, NY, and Kriegler, 1990; GENE TRANSFER AND EXPRESSION, A LABORATORY MANUAL, Stockton Press, NY.

[0063] In a third embodiment, a nucleic acid that is hybridizable to the nucleic acid molecule comprising the nucleotide sequences of SEQ ID NOS:1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, 75, 77, 79, 81, 83, 85, 87, 89, 91, 93, 95, 97, 99, 101, 103, 105, 107, 109, 111, 113, 115, 117, 119, 121, 123, 125, 127, 129, 131, 133, 135, 137, 139, 141, 143, 145, 147, 149 and 151 or fragments, analogs or derivatives thereof, under conditions of low stringency, is provided. A non-limiting example of low stringency hybridization conditions are hybridization in 35% formamide, 5×SSC, 50 mM Tris-HCl (pH 7.5), 5 mM EDTA, 0.02% PVP, 0.02% Ficoll, 0.2% BSA, 100 mg/ml denatured salmon sperm DNA, 10% (wt/vol) dextran sulfate at 40° C., followed by one or more washes in 2×SSC, 25 mM Tris-HCl (pH 7.4), 5 mM EDTA, and 0.1% SDS at 50° C. Other conditions of low stringency that may be used are well known in the art (e.g., as employed for cross-species hybridizations). See, e.g., Ausubel, et al. (eds.), 1993, CURRENT PROTOCOLS IN MOLECULAR BIOLOGY, John Wiley & Sons, NY, and Kriegler, 1990, GENE TRANSFER AND EXPRESSION, A LABORATORY MANUAL, Stockton Press, NY; Shilo and Weinberg, 1981. Proc Natl Acad Sci USA 78: 6789-6792.

[0064] Conservative Mutations

[0065] In addition to naturally-occurring allelic variants of GPCRX sequences that may exist in the population, the skilled artisan will further appreciate that changes can be introduced by mutation into the nucleotide sequences of SEQ ID NOS:1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, 75, 77, 79, 81, 83, 85, 87, 89, 91, 93, 95, 97, 99, 101, 103, 105, 107, 109, 111, 113, 115, 117, 119, 121, 123, 125, 127, 129, 131, 133, 135, 137, 139, 141, 143, 145, 147, 149 and 151 thereby leading to changes in the amino acid sequences of the encoded GPCRX proteins, without altering the functional ability of said GPCRX proteins. For example, nucleotide substitutions leading to amino acid substitutions at “non-essential” amino acid residues can be made in the sequence of SEQ ID NOS: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, 78, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 100, 102, 104, 106, 108, 110, 112, 114, 116, 118, 120, 122, 124, 126, 128, 130, 132, 134, 136, 138, 140, 142, 144, 146, 148, 150 and 152. A “non-essential” amino acid residue is a residue that can be altered from the wild-type sequences of the GPCRX proteins without altering their biological activity, whereas an “essential” amino acid residue is required for such biological activity. For example, amino acid residues that are conserved among the GPCRX proteins of the invention are predicted to be particularly non-amenable to alteration. Amino acids for which conservative substitutions can be made are well-known within the art.

[0066] Another aspect of the invention pertains to nucleic acid molecules encoding GPCRX proteins that contain changes in amino acid residues that are not essential for activity. Such GPCRX proteins differ in amino acid sequence from SEQ ID NOS: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, 78, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 100, 102, 104, 106, 108, 110, 112, 114, 116, 118, 120, 122, 124, 126, 128, 130, 132, 134, 136, 138, 140, 142, 144, 146, 148, 150 and 152 yet retain biological activity. In one embodiment, the isolated nucleic acid molecule comprises a nucleotide sequence encoding a protein, wherein the protein comprises an amino acid sequence at least about 45% homologous to the amino acid sequences of SEQ ID NOS: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, 78, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 100, 102, 104, 106, 108, 110, 112, 114, 116, 118, 120, 122, 124, 126, 128, 130, 132, 134, 136, 138, 140, 142, 144, 146, 148, 150 and 152. Preferably, the protein encoded by the nucleic acid molecule is at least about 60% homologous to SEQ ID NOS: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, 78, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 100, 102, 104, 106, 108, 110, 112, 114, 116, 118, 120, 122, 124, 126, 128, 130, 132, 134, 136, 138, 140, 142, 144, 146, 148, 150 and 152; more preferably at least about 70% homologous to SEQ ID NOS: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, 78, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 100, 102, 104, 106, 108, 110, 112, 114, 116, 118, 120, 122, 124, 126, 128, 130, 132, 134, 136, 138, 140, 142, 144, 146, 148, 150 and 152; still more preferably at least about 80% homologous to SEQ ID NOS: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, 78, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 100, 102, 104, 106, 108, 110, 112, 114, 116, 118, 120, 122, 124, 126, 128, 130, 132, 134, 136, 138, 140, 142, 144, 146, 148, 150 and 152; even more preferably at least about 90% homologous to SEQ ID NOS: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, 78, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 100, 102, 104, 106, 108, 110, 112, 114, 116, 118, 120, 122, 124, 126, 128, 130, 132, 134, 136, 138, 140, 142, 144, 146, 148, 150 and 152; and most preferably at least about 95% homologous to SEQ ID NOS: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, 78, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 100, 102, 104, 106, 108, 110, 112, 114, 116, 118, 120, 122, 124, 126, 128, 130, 132, 134, 136, 138, 140, 142, 144, 146, 148, 150 and 152.

[0067] An isolated nucleic acid molecule encoding an GPCRX protein homologous to the protein of SEQ ID NOS: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, 78, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 100, 102, 104, 106, 108, 110, 112, 114, 116, 118, 120, 122, 124, 126, 128, 130, 132, 134, 136, 138, 140, 142, 144, 146, 148, 150 and 152 can be created by introducing one or more nucleotide substitutions, additions or deletions into the nucleotide sequence of SEQ ID NOS:1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, 75, 77, 79, 81, 83, 85, 87, 89, 91, 93, 95, 97, 99, 101, 103, 105, 107, 109, 111, 113, 115, 117, 119, 121, 123, 125, 127, 129, 131, 133, 135, 137, 139, 141, 143, 145, 147, 149 and 151 such that one or more amino acid substitutions, additions or deletions are introduced into the encoded protein.

[0068] Mutations can be introduced into SEQ ID NOS: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, 78, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 100, 102, 104, 106, 108, 110, 112, 114, 116, 118, 120, 122, 124, 126, 128, 130, 132, 134, 136, 138, 140, 142, 144, 146, 148, 150 and 152 by standard techniques, such as site-directed mutagenesis and PCR-mediated mutagenesis. Preferably, conservative amino acid substitutions are made at one or more predicted, non-essential amino acid residues. A “conservative amino acid substitution” is one in which the amino acid residue is replaced with an amino acid residue having a similar side chain. Families of amino acid residues having similar side chains have been defined within the art. These families include amino acids with basic side chains (e.g., lysine, arginine, histidine), acidic side chains (e.g., aspartic acid, glutamic acid), uncharged polar side chains (e.g., glycine, asparagine, glutamine, serine, threonine, tyrosine, cysteine), nonpolar side chains (e.g., alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine, tryptophan), beta-branched side chains (e.g., threonine, valine, isoleucine) and aromatic side chains (e.g., tyrosine, phenylalanine, tryptophan, histidine). Thus, a predicted non-essential amino acid residue in the GPCRX protein is replaced with another amino acid residue from the same side chain family. Alternatively, in another embodiment, mutations can be introduced randomly along all or part of an GPCRX coding sequence, such as by saturation mutagenesis, and the resultant mutants can be screened for GPCRX biological activity to identify mutants that retain activity. Following mutagenesis of SEQ ID NOS:1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, 75, 77, 79, 81, 83, 85, 87, 89, 91, 93, 95, 97, 99, 101, 103, 105, 107, 109, 111, 113, 115, 117, 119, 121, 123, 125, 127, 129, 131, 133, 135, 137, 139, 141, 143, 145, 147, 149 and 151, the encoded protein can be expressed by any recombinant technology known in the art and the activity of the protein can be determined.

[0069] The relatedness of amino acid families may also be determined based on side chain interactions. Substituted amino acids may be fully conserved “strong” residues or fully conserved “weak” residues. The “strong” group of conserved amino acid residues may be any one of the following groups: STA, NEQK, NHQK, NDEQ, QHRK, MILV, MILF, HY, FYW, wherein the single letter amino acid codes are grouped by those amino acids that may be substituted for each other. Likewise, the “weak” group of conserved residues may be any one of the following: CSA, ATV, SAG, STNK, STPA, SGND, SNDEQK, NDEQHK, NEQHRK, VLIM, HFY, wherein the letters within each group represent the single letter amino acid code.

[0070] In one embodiment, a mutant GPCRX protein can be assayed for (i) the ability to form protein:protein interactions with other GPCRX proteins, other cell-surface proteins, or biologically-active portions thereof, (ii) complex formation between a mutant GPCRX protein and an GPCRX ligand; or (iii) the ability of a mutant GPCRX protein to bind to an intracellular target protein or biologically-active portion thereof; (e.g. avidin proteins).

[0071] In yet another embodiment, a mutant GPCRX protein can be assayed for the ability to regulate a specific biological function (e.g., regulation of insulin release). Antisense Nucleic Acids Another aspect of the invention pertains to isolated antisense nucleic acid molecules that are hybridizable to or complementary to the nucleic acid molecule comprising the nucleotide sequence of SEQ ID NOS:1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, 75, 77, 79, 81, 83, 85, 87, 89, 91, 93, 95, 97, 99, 101, 103, 105, 107, 109, 111, 113, 115, 117, 119, 121, 123, 125, 127, 129, 131, 133, 135, 137, 139, 141, 143, 145, 147, 149 and 151, or fragments, analogs or derivatives thereof. An “antisense” nucleic acid comprises a nucleotide sequence that is complementary to a “sense” nucleic acid encoding a protein (e.g., complementary to the coding strand of a double-stranded cDNA molecule or complementary to an mRNA sequence). In specific aspects, antisense nucleic acid molecules are provided that comprise a sequence complementary to at least about 10, 25, 50, 100, 250 or 500 nucleotides or an entire GPCRX coding strand, or to only a portion thereof. Nucleic acid molecules encoding fragments, homologs, derivatives and analogs of an GPCRX protein of SEQ ID NOS: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, 78, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 100, 102, 104, 106, 108, 110, 112, 114, 116, 118, 120, 122, 124, 126, 128, 130, 132, 134, 136, 138, 140, 142, 144, 146, 148, 150 and 152, or antisense nucleic acids complementary to an GPCRX nucleic acid sequence of SEQ ID NOS:1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, 75, 77, 79, 81, 83, 85, 87, 89, 91, 93, 95, 97, 99, 101, 103, 105, 107, 109, 111, 113, 115, 117, 119, 121, 123, 125, 127, 129, 131, 133, 135, 137, 139, 141, 143, 145, 147, 149 and 151, are additionally provided.

[0072] In one embodiment, an antisense nucleic acid molecule is antisense to a “coding region” of the coding strand of a nucleotide sequence encoding an GPCRX protein. The term “coding region” refers to the region of the nucleotide sequence comprising codons which are translated into amino acid residues. In another embodiment, the antisense nucleic acid molecule is antisense to a “noncoding region” of the coding strand of a nucleotide sequence encoding the GPCRX protein. The term “noncoding region” refers to 5′ and 3′ sequences which flank the coding region that are not translated into amino acids (i.e., also referred to as 5′ and 3′ untranslated regions).

[0073] Given the coding strand sequences encoding the GPCRX protein disclosed herein, antisense nucleic acids of the invention can be designed according to the rules of Watson and Crick or Hoogsteen base pairing. The antisense nucleic acid molecule can be complementary to the entire coding region of GPCRX mRNA, but more preferably is an oligonucleotide that is antisense to only a portion of the coding or noncoding region of GPCRX mRNA. For example, the antisense oligonucleotide can be complementary to the region surrounding the translation start site of GPCRX mRNA. An antisense oligonucleotide can be, for example, about 5, 10, 15, 20, 25, 30, 35, 40, 45 or 50 nucleotides in length. An antisense nucleic acid of the invention can be constructed using chemical synthesis or enzymatic ligation reactions using procedures known in the art. For example, an antisense nucleic acid (e.g., an antisense oligonucleotide) can be chemically synthesized using naturally-occurring nucleotides or variously modified nucleotides designed to increase the biological stability of the molecules or to increase the physical stability of the duplex formed between the antisense and sense nucleic acids (e.g., phosphorothioate derivatives and acridine substituted nucleotides can be used).

[0074] Examples of modified nucleotides that can be used to generate the antisense nucleic acid include: 5-fluorouracil, 5-bromouracil, 5-chlorouracil, 5-iodouracil, hypoxanthine, xanthine, 4-acetylcytosine, 5-(carboxyhydroxylmethyl) uracil, 5-carboxymethylaminomethyl-2-thiouridine, 5-carboxymethylaminomethyluracil, dihydrouracil, beta-D-galactosylqueosine, inosine, N6-isopentenyladenine, 1-methylguanine, 1-methylinosine, 2,2-dimethylguanine, 2-methyladenine, 2-methylguanine, 3-methylcytosine, 5-methylcytosine, N6-adenine, 7-methylguanine, 5-methylaminomethyluracil, 5-methoxyaminomethyl-2-thiouracil, beta-D-mannosylqueosine, 5′-methoxycarboxymethyluracil, 5-methoxyuracil, 2-methylthio-N-6-isopentenyladenine, uracil-5-oxyacetic acid (v), wybutoxosine, pseudouracil, queosine, 2-thiocytosine, 5-methyl-2-thiouracil, 2-thiouracil, 4-thiouracil, 5-methyluracil, uracil-5-oxyacetic acid methylester, uracil-5-oxyacetic acid (v), 5-methyl-2-thiouracil, 3-(3-amino-3-N-2-carboxypropyl) uracil, (acp3)w, and 2,6-diaminopurine. Alternatively, the antisense nucleic acid can be produced biologically using an expression vector into which a nucleic acid has been subcloned in an antisense orientation (i.e., RNA transcribed from the inserted nucleic acid will be of an antisense orientation to a target nucleic acid of interest, described further in the following subsection).

[0075] The antisense nucleic acid molecules of the invention are typically administered to a subject or generated in situ such that they hybridize with or bind to cellular mRNA and/or genomic DNA encoding an GPCRX protein to thereby inhibit expression of the protein (e.g., by inhibiting transcription and/or translation). The hybridization can be by conventional nucleotide complementarity to form a stable duplex, or, for example, in the case of an antisense nucleic acid molecule that binds to DNA duplexes, through specific interactions in the major groove of the double helix. An example of a route of administration of antisense nucleic acid molecules of the invention includes direct injection at a tissue site. Alternatively, antisense nucleic acid molecules can be modified to target selected cells and then administered systemically. For example, for systemic administration, antisense molecules can be modified such that they specifically bind to receptors or antigens expressed on a selected cell surface (e.g., by linking the antisense nucleic acid molecules to peptides or antibodies that bind to cell surface receptors or antigens). The antisense nucleic acid molecules can also be delivered to cells using the vectors described herein. To achieve sufficient nucleic acid molecules, vector constructs in which the antisense nucleic acid molecule is placed under the control of a strong pol II or pol III promoter are preferred.

[0076] In yet another embodiment, the antisense nucleic acid molecule of the invention is an α-anomeric nucleic acid molecule. An α-anomeric nucleic acid molecule forms specific double-stranded hybrids with complementary RNA in which, contrary to the usual β-units, the strands run parallel to each other. See, e.g., Gaultier, et al., 1987. Nucl. Acids Res. 15: 6625-6641. The antisense nucleic acid molecule can also comprise a 2′-o-methylribonucleotide (see, e.g., Inoue, et al. 1987. Nucl. Acids Res. 15: 6131-6148) or a chimeric RNA-DNA analogue (see, e.g., Inoue, et al., 1987. FEBS Lett. 215: 327-330.

[0077] Ribozymes and PNA Moieties

[0078] Nucleic acid modifications include, by way of non-limiting example, modified bases, and nucleic acids whose sugar phosphate backbones are modified or derivatized. These modifications are carried out at least in part to enhance the chemical stability of the modified nucleic acid, such that they may be used, for example, as antisense binding nucleic acids in therapeutic applications in a subject.

[0079] In one embodiment, an antisense nucleic acid of the invention is a ribozyme. -Ribozymes are catalytic RNA molecules with ribonuclease activity that are capable of cleaving a single-stranded nucleic acid, such as an mRNA, to which they have a complementary region. Thus, ribozymes (e.g., hammerhead ribozymes as described in Haselhoff and Gerlach 1988. Nature 334: 585-591) can be used to catalytically cleave GPCRX mRNA transcripts to thereby inhibit translation of GPCRX mRNA. A ribozyme having specificity for an GPCRX-encoding nucleic acid can be designed based upon the nucleotide sequence of an GPCRX cDNA disclosed herein (i.e., SEQ ID NOS:1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, 75, 77, 79, 81, 83, 85, 87, 89, 91, 93, 95, 97, 99, 101, 103, 105, 107, 109, 111, 113, 115, 117, 119, 121, 123, 125, 127, 129, 131, 133, 135, 137, 139, 141, 143, 145, 147, 149 and 151). For example, a derivative of a Tetrahymena L-19 IVS RNA can be constructed in which the nucleotide sequence of the active site is complementary to the nucleotide sequence to be cleaved in an GPCRX-encoding mRNA. See, e.g., U.S. Pat. No. 4,987,071 to Cech, et al. and U.S. Pat. No. 5,116,742 to Cech, et al. GPCRX mRNA can also be used to select a catalytic RNA having a specific ribonuclease activity from a pool of RNA molecules. See, e.g., Bartel et al., (1993) Science 261:1411-1418.

[0080] Alternatively, GPCRX gene expression can be inhibited by targeting nucleotide sequences complementary to the regulatory region of the GPCRX nucleic acid (e.g. the GPCRX promoter and/or enhancers) to form triple helical structures that prevent transcription of the GPCRX gene in target cells. See, e.g., Helene, 1991. Anticancer Drug Des. 6: 569-84; Helene, et al. 1992. Ann. N.Y. Acad. Sci. 660: 27-36; Maher, 1992. Bioassays 14: 807-15.

[0081] In various embodiments, the GPCRX nucleic acids can be modified at the base moiety, sugar moiety or phosphate backbone to improve, e.g., the stability, hybridization, or solubility of the molecule. For example, the deoxyribose phosphate backbone of the nucleic acids can be modified to generate peptide nucleic acids. See, e.g., Hyrup, et al., 1996. Bioorg Med Chem 4: 5-23. As used herein, the terms “peptide nucleic acids” or “PNAs” refer to nucleic acid mimics (e.g., DNA mimics) in which the deoxyribose phosphate backbone is replaced by a pseudopeptide backbone and only the four natural nucleobases are retained. The neutral backbone of PNAs has been shown to allow for specific hybridization to DNA and RNA under conditions of low ionic strength. The synthesis of PNA oligomers can be performed using standard solid phase peptide synthesis protocols as described in Hyrup, et al., 1996. supra; Perry-O'Keefe, et al., 1996. Proc. Natl. Acad. Sci. USA 93: 14670-14675.

[0082] PNAs of GPCRX can be used in therapeutic and diagnostic applications. For example, PNAs can be used as antisense or antigene agents for sequence-specific modulation of gene expression by, e.g., inducing transcription or translation arrest or inhibiting replication. PNAs of GPCRX can also be used, for example, in the analysis of single base pair mutations in a gene (e.g., PNA directed PCR clamping; as artificial restriction enzymes when used in combination with other enzymes, e.g., S1 nucleases (see, Hyrup, et al., 1996.supra); or as probes or primers for DNA sequence and hybridization (see, Hyrup, et al., 1996, supra; Perry-O'Keefe, et al., 1996. supra).

[0083] In another embodiment, PNAs of GPCRX can be modified, e.g., to enhance their stability or cellular uptake, by attaching lipophilic or other helper groups to PNA, by the formation of PNA-DNA chimeras, or by the use of liposomes or other techniques of drug delivery known in the art. For example, PNA-DNA chimeras of GPCRX can be generated that may combine the advantageous properties of PNA and DNA. Such chimeras allow DNA recognition enzymes (e.g., RNase H and DNA polymerases) to interact with the DNA portion while the PNA portion would provide high binding affinity and specificity. PNA-DNA chimeras can be linked using linkers of appropriate lengths selected in terms of base stacking, number of bonds between the nucleobases, and orientation (see, Hyrup, et al., 1996. supra). The synthesis of PNA-DNA chimeras can be performed as described in Hyrup, et al., 1996. supra and Finn, et al., 1996. Nucl Acids Res 24: 3357-3363. For example, a DNA chain can be synthesized on a solid support using standard phosphoramidite coupling chemistry, and modified nucleoside analogs, e.g., 5′-(4-methoxytrityl)amino-5′-deoxy-thymidine phosphoramidite, can be used between the PNA and the 5′ end of DNA. See, e.g., Mag, et al., 1989. Nucl Acid Res 17: 5973-5988. PNA monomers are then coupled in a stepwise manner to produce a chimeric molecule with a 5′ PNA segment and a 3′ DNA segment. See, e.g., Finn, et al., 1996. supra. Alternatively, chimeric molecules can be synthesized with a 5′ DNA segment and a 3′ PNA segment. See, e.g., Petersen, et al., 1975. Bioorg. Med. Chem. Lett. 5: 1119-11124.

[0084] In other embodiments, the oligonucleotide may include other appended groups such as -peptides (e.g., for targeting host cell receptors in vivo), or agents facilitating transport across the cell membrane (see, e.g., Letsinger, et al., 1989. Proc. Natl. Acad. Sci. U.S.A. 86: 6553-6556; Lemaitre, et al., 1987. Proc. Natl. Acad. Sci. 84: 648-652; PCT Publication No. WO88/09810) or the blood-brain barrier (see, e.g., PCT Publication No. WO 89/10134). In addition, oligonucleotides can be modified with hybridization triggered cleavage agents (see, e.g., Krol, et al., 1988. BioTechniques 6:958-976) or intercalating agents (see, e.g. Zon, 1988. Pharm. Res. 5: 539-549). To this end, the oligonucleotide may be conjugated to another molecule, e.g., a peptide, a hybridization triggered cross-linking agent, a transport agent, a hybridization-triggered cleavage agent, and the like.

[0085] GPCRX Polypeptides

[0086] A polypeptide according to the invention includes a polypeptide including the amino acid sequence of GPCRX polypeptides whose sequences are provided in SEQ ID NOS: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, 78, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 100, 102, 104, 106, 108, 110, 112, 114, 116, 118, 120, 122, 124, 126, 128, 130, 132, 134, 136, 138, 140, 142, 144, 146, 148, 150 and 152. The invention also includes a mutant or variant protein any of whose residues may be changed from the corresponding residues shown in SEQ ID NOS: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, 78, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 100, 102, 104, 106, 108, 110, 112, 114, 116, 118, 120, 122, 124, 126, 128, 130, 132, 134, 136, 138, 140, 142, 144, 146, 148, 150 and 152 while still encoding a protein that maintains its GPCRX activities and physiological functions, or a functional fragment thereof.

[0087] In general, an GPCRX variant that preserves GPCRX-like function includes any variant in which residues at a particular position in the sequence have been substituted by other amino acids, and further include the possibility of inserting an additional residue or residues between two residues of the parent protein as well as the possibility of deleting one or more residues from the parent sequence. Any amino acid substitution, insertion, or deletion is encompassed by the invention. In favorable circumstances, the substitution is a conservative substitution as defined above.

[0088] One aspect of the invention pertains to isolated GPCRX proteins, and biologically-active portions thereof, or derivatives, fragments, analogs or homologs thereof. Also provided are polypeptide fragments suitable for use as immunogens to raise anti-GPCRX antibodies. In one embodiment, native GPCRX proteins can be isolated from cells or tissue sources by an appropriate purification scheme using standard protein purification techniques. In another embodiment, GPCRX proteins are produced by recombinant DNA techniques. Alternative to recombinant expression, an GPCRX protein or polypeptide can be synthesized chemically using standard peptide synthesis techniques.

[0089] An “isolated” or “purified” polypeptide or protein or biologically-active portion thereof is substantially free of cellular material or other contaminating proteins from the cell or tissue source from which the GPCRX protein is derived, or substantially free from chemical precursors or other chemicals when chemically synthesized. The language “substantially free of cellular material” includes preparations of GPCRX proteins in which the protein is separated from cellular components of the cells from which it is isolated or recombinantly-produced. In one embodiment, the language “substantially free of cellular material” includes preparations of GPCRX proteins having less than about 30% (by dry weight) of non-GPCRX proteins (also referred to herein as a “contaminating protein”), more preferably less than about 20% of non-GPCRX proteins, still more preferably less than about 10% of non-GPCRX proteins, and most preferably less than about 5% of non-GPCRX proteins. When the GPCRX protein or biologically-active portion thereof is recombinantly-produced, it is also preferably substantially free of culture medium, i.e., culture medium represents less than about 20%, more preferably less than about 10%, and most preferably less than about 5% of the volume of the GPCRX protein preparation.

[0090] The language “substantially free of chemical precursors or other chemicals” includes preparations of GPCRX proteins in which the protein is separated from chemical precursors or other chemicals that are involved in the synthesis of the protein. In one embodiment, the language “substantially free of chemical precursors or other chemicals” includes preparations of GPCRX proteins having less than about 30% (by dry weight) of chemical precursors or non-GPCRX chemicals, more preferably less than about 20% chemical precursors or non-GPCRX chemicals, still more preferably less than about 10% chemical precursors or non-GPCRX chemicals, and most preferably less than about 5% chemical precursors or non-GPCRX chemicals.

[0091] Biologically-active portions of GPCRX proteins include peptides comprising amino acid sequences sufficiently homologous to or derived from the amino acid sequences of the GPCRX proteins (e.g., the amino acid sequence shown in SEQ ID NOS: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, 78, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 100, 102, 104, 106, 108, 110, 112, 114, 116, 118, 120, 122, 124, 126, 128, 130, 132, 134, 136, 138, 140, 142, 144, 146, 148, 150 and 152) that include fewer amino acids than the full-length GPCRX proteins, and exhibit at least one activity of an GPCRX protein. Typically, biologically-active portions comprise a domain or motif with at least one activity of the GPCRX protein. A biologically-active portion of an GPCRX protein can be a polypeptide which is, for example, 10, 25, 50, 100 or more amino acid residues in length.

[0092] Moreover, other biologically-active portions, in which other regions of the protein are deleted, can be prepared by recombinant techniques and evaluated for one or more of the functional activities of a native GPCRX protein.

[0093] In an embodiment, the GPCRX protein has an amino acid sequence shown in SEQ ID NOS: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, 78, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 100, 102, 104, 106, 108, 110, 112, 114, 116, 118, 120, 122, 124, 126, 128, 130, 132, 134, 136, 138, 140, 142, 144, 146, 148, 150 and 152. In other embodiments, the GPCRX protein is substantially homologous to SEQ ID NOS: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, 78, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 100, 102, 104, 106, 108, 110, 112, 114, 116, 118, 120, 122, 124, 126, 128, 130, 132, 134, 136, 138, 140, 142, 144, 146, 148, 150 and 152, and retains the functional activity of the protein of SEQ ID NOS: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, 78, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 100, 102, 104, 106, 108, 110, 112, 114, 116, 118, 120, 122, 124, 126, 128, 130, 132, 134, 136, 138, 140, 142, 144, 146, 148, 150 and 152, yet differs in amino acid sequence due to natural allelic variation or mutagenesis, as described in detail, below. Accordingly, in another embodiment, the GPCRX protein is a protein that comprises an amino acid sequence at least about 45% homologous to the amino acid sequence SEQ ID NOS: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, 78, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 100, 102, 104, 106, 108, 110, 112, 114, 116, 118, 120, 122, 124, 126, 128, 130, 132, 134, 136, 138, 140, 142, 144, 146, 148, 150 and 152, and retains the functional activity of the GPCRX proteins of SEQ ID NOS: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, 78, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 100, 102, 104, 106, 108, 110, 112, 114, 116, 118, 120, 122, 124, 126, 128, 130, 132, 134, 136, 138, 140, 142, 144, 146, 148, 150 and 152.

[0094] Determining Homology Between Two or More Sequences

[0095] To determine the percent homology of two amino acid sequences or of two nucleic acids, the sequences are aligned for optimal comparison purposes (e.g., gaps can be introduced in the sequence of a first amino acid or nucleic acid sequence for optimal alignment with a second amino or nucleic acid sequence). The amino acid residues or nucleotides at corresponding amino acid positions or nucleotide positions are then compared. When a position in the first sequence is occupied by the same amino acid residue or nucleotide as the corresponding position in the second sequence, then the molecules are homologous at that—position (i.e., as used herein amino acid or nucleic acid “homology” is equivalent to amino acid or nucleic acid “identity”).

[0096] The nucleic acid sequence homology may be determined as the degree of identity between two sequences. The homology may be determined using computer programs known in the art, such as GAP software provided in the GCG program package. See, Needleman and Wunsch, 1970. J Mol Biol 48: 443-453. Using GCG GAP software with the following settings for nucleic acid sequence comparison: GAP creation penalty of 5.0 and GAP extension penalty of 0.3, the coding region of the analogous nucleic acid sequences referred to above exhibits a degree of identity preferably of at least 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99%, with the CDS (encoding) part of the DNA sequence shown in SEQ ID NOS:1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, 75, 77, 79, 81, 83, 85, 87, 89, 91, 93, 95, 97, 99, 101, 103, 105, 107, 109, 111, 113, 115, 117, 119, 121, 123, 125, 127, 129, 131, 133, 135, 137, 139, 141, 143, 145, 147, 149 and 151.

[0097] The term “sequence identity” refers to the degree to which two polynucleotide or polypeptide sequences are identical on a residue-by-residue basis over a particular region of comparison. The term “percentage of sequence identity” is calculated by comparing two optimally aligned sequences over that region of comparison, determining the number of positions at which the identical nucleic acid base (e.g., A, T, C, G, U, or I, in the case of nucleic acids) occurs in both sequences to yield the number of matched positions, dividing the number of matched positions by the total number of positions in the region of comparison (i.e., the window size), and multiplying the result by 100 to yield the percentage of sequence identity. The term “substantial identity” as used herein denotes a characteristic of a polynucleotide sequence, wherein the polynucleotide comprises a sequence that has at least 80 percent sequence identity, preferably at least 85 percent identity and often 90 to 95 percent sequence identity, more usually at least 99 percent sequence identity as compared to a reference sequence over a comparison region.

[0098] Chimeric and Fusion Proteins

[0099] The invention also provides GPCRX chimeric or fusion proteins. As used herein, an GPCRX “chimeric protein” or “fusion protein” comprises an GPCRX polypeptide operatively-linked to a non-GPCRX polypeptide. An “GPCRX polypeptide” refers to a polypeptide having an amino acid sequence corresponding to an GPCRX protein (SEQ ID NOS: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, 78, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 100, 102, 104, 106, 108, 110, 112, 114, 116, 118, 120, 122, 124, 126, 128, 130, 132, 134, 136, 138, 140, 142, 144, 146, 148, 150 and 152), whereas a “non-GPCRX polypeptide” refers to a polypeptide having an amino acid sequence corresponding to a protein that is not substantially homologous to the GPCRX protein, e.g., a protein that is different from the GPCRX protein and that is derived from the same or a different organism. Within an GPCRX fusion protein the GPCRX polypeptide can correspond to all or a portion of an GPCRX protein. In one embodiment, an GPCRX fusion protein comprises at least one biologically-active portion of an GPCRX protein. In another embodiment, an GPCRX fusion protein comprises at least two biologically-active portions of an GPCRX protein. In yet another embodiment, an GPCRX fusion protein comprises at least three biologically-active portions of an GPCRX protein. Within the fusion protein, the term “operatively-linked” is intended to indicate that the GPCRX polypeptide and the non-GPCRX polypeptide are fused in-frame with one another. The non-GPCRX polypeptide can be fused to the N-terminus or C-terminus of the GPCRX polypeptide.

[0100] In one embodiment, the fusion protein is a GST-GPCRX fusion protein in which the GPCRX sequences are fused to the C-terminus of the GST (glutathione S-transferase) sequences. Such fusion proteins can facilitate the purification of recombinant GPCRX polypeptides.

[0101] In another embodiment, the fusion protein is an GPCRX protein containing a heterologous signal sequence at its N-terminus. In certain host cells (e.g., mammalian host cells), expression and/or secretion of GPCRX can be increased through use of a heterologous signal sequence.

[0102] In yet another embodiment, the fusion protein is an GPCRX-immunoglobulin fusion protein in which the GPCRX sequences are fused to sequences derived from a member of the immunoglobulin protein family. The GPCRX-immunoglobulin fusion proteins of the invention can be incorporated into pharmaceutical compositions and administered to a subject to inhibit an interaction between an GPCRX ligand and an GPCRX protein on the surface of a cell, to thereby suppress GPCRX-mediated signal transduction in vivo. The GPCRX-immunoglobulin fusion proteins can be used to affect the bioavailability of an GPCRX cognate ligand. Inhibition of the GPCRX ligand/GPCRX interaction may be useful therapeutically for both the treatment of proliferative and differentiative disorders, as well as modulating (e.g. promoting or inhibiting) cell survival. Moreover, the GPCRX-immunoglobulin fusion proteins of the invention can be used as immunogens to produce anti-GPCRX antibodies in a subject, to purify GPCRX ligands, and in screening assays to identify molecules that inhibit the interaction of GPCRX with an GPCRX ligand.

[0103] An GPCRX chimeric or fusion protein of the invention can be produced by standard recombinant DNA techniques. For example, DNA fragments coding for the different polypeptide sequences are ligated together in-frame in accordance with conventional techniques, e.g., by employing blunt-ended or stagger-ended termini for ligation, restriction enzyme digestion to provide for appropriate termini, filling-in of cohesive ends as appropriate, alkaline phosphatase treatment to avoid undesirable joining, and enzymatic ligation. In another embodiment, the fusion gene can be synthesized by conventional techniques including automated DNA synthesizers. Alternatively, PCR amplification of gene fragments can be carried out using anchor primers that give rise to complementary overhangs between two consecutive gene fragments that can subsequently be annealed and reamplified to generate a chimeric gene sequence (see, e.g., Ausubel, et al. (eds.) CURRENT PROTOCOLS IN MOLECULAR BIOLOGY, John Wiley & Sons, 1992). Moreover, many expression vectors are commercially available that already encode a fusion moiety (e.g., a GST polypeptide). An GPCRX-encoding nucleic acid can be cloned into such an expression vector such that the fusion moiety is linked in-frame to the GPCRX protein.

[0104] GPCRX Agonists and Antagonists

[0105] The invention also pertains to variants of the GPCRX proteins that function as either GPCRX agonists (i.e., mimetics) or as GPCRX antagonists. Variants of the GPCRX protein can be generated by mutagenesis (e.g., discrete point mutation or truncation of the GPCRX protein). An agonist of the GPCRX protein can retain substantially the same, or a subset of, the biological activities of the naturally occurring form of the GPCRX protein. An antagonist of the GPCRX protein can inhibit one or more of the activities of the naturally occurring form of the GPCRX protein by, for example, competitively binding to a downstream or upstream member of a cellular signaling cascade which includes the GPCRX protein. Thus, specific biological effects can be elicited by treatment with a variant of limited function. In one embodiment, treatment of a subject with a variant having a subset of the biological activities of the naturally occurring form of the protein has fewer side effects in a subject relative to treatment with the naturally occurring form of the GPCRX proteins.

[0106] Variants of the GPCRX proteins that function as either GPCRX agonists (i.e., mimetics) or as GPCRX antagonists can be identified by screening combinatorial libraries of mutants (e.g., truncation mutants) of the GPCRX proteins for GPCRX protein agonist or antagonist activity. In one embodiment, a variegated library of GPCRX variants is generated by combinatorial mutagenesis at the nucleic acid level and is encoded by a variegated gene library. A variegated library of GPCRX variants can be produced by, for example, enzymatically ligating a mixture of synthetic oligonucleotides into gene sequences such that a degenerate set of potential GPCRX sequences is expressible as individual polypeptides, or alternatively, as a set of larger fusion proteins (e.g., for phage display) containing the set of GPCRX sequences therein. There are a variety of methods which can be used to produce libraries of potential GPCRX variants from a degenerate oligonucleotide sequence. Chemical synthesis of a degenerate gene sequence can be performed in an automatic DNA synthesizer, and the synthetic gene then ligated into an appropriate expression vector. Use of a degenerate set of genes allows for the provision, in one mixture, of all of the sequences encoding the desired set of potential GPCRX sequences. Methods for synthesizing degenerate oligonucleotides are well-known within the art. See, e.g., Narang, 1983. Tetrahedron 39: 3; Itakura, et al., 1984. Annu. Rev. Biochem. 53: 323; Itakura, et al., 1984. Science 198: 1056; Ike, et al., 1983. Nucl. Acids Res. 11: 477.

[0107] Polypeptide Libraries

[0108] In addition, libraries of fragments of the GPCRX protein coding sequences can be used to generate a variegated population of GPCRX fragments for screening and subsequent selection of variants of an GPCRX protein. In one embodiment, a library of coding sequence fragments can be generated by treating a double stranded PCR fragment of an GPCRX coding sequence with a nuclease under conditions wherein nicking occurs only about once per molecule, denaturing the double stranded DNA, renaturing the DNA to form double-stranded DNA that can include sense/antisense pairs from different nicked products, removing single stranded portions from reformed duplexes by treatment with S1 nuclease, and ligating the resulting fragment library into an expression vector. By this method, expression libraries can be derived which encodes N-terminal and internal fragments of various sizes of the GPCRX proteins.

[0109] Various techniques are known in the art for screening gene products of combinatorial libraries made by point mutations or truncation, and for screening cDNA libraries for gene products having a selected property. Such techniques are adaptable for rapid screening of the gene libraries generated by the combinatorial mutagenesis of GPCRX proteins. The most widely used techniques, which are amenable to high throughput analysis, for screening large gene libraries typically include cloning the gene library into replicable expression vectors, transforming appropriate cells with the resulting library of vectors, and expressing the combinatorial genes under conditions in which detection of a desired activity facilitates isolation of the vector encoding the gene whose product was detected. Recursive ensemble mutagenesis (REM), a new technique that enhances the frequency of functional mutants in the libraries, can be used in combination with the screening assays to identify GPCRX variants. See, e.g., Arkin and Yourvan, 1992. Proc. Natl. Acad. Sci. USA 89: 7811-7815; Delgrave, et al., 1993. Protein Engineering 6:327-331.

[0110] Anti-GPCRX Antibodies

[0111] Also included in the invention are antibodies to GPCRX proteins, or fragments of GPCRX proteins. The term “antibody” as used herein refers to immunoglobulin molecules and immunologically active portions of immunoglobulin (Ig) molecules, i.e., molecules that contain an antigen binding site that specifically binds (immunoreacts with) an antigen. Such antibodies include, but are not limited to, polyclonal, monoclonal, chimeric, single chain, Fab, Fab′ and F(ab′)2 fragments, and an Fab expression library. In general, an antibody molecule obtained from humans relates to any of the classes IgG, IgM, IgA, IgE and IgD, which differ from one another by the nature of the heavy chain present in the molecule. Certain classes have subclasses as well, such as IgG1, IgG2, and others. Furthermore, in humans, the light chain may be a kappa chain or a lambda chain. Reference herein to antibodies includes a reference to all such classes, subclasses and types of human antibody species.

[0112] An isolated GPCRX-related protein of the invention may be intended to serve as an antigen, or a portion or fragment thereof, and additionally can be used as an immunogen to generate antibodies that immunospecifically bind the antigen, using standard techniques for polyclonal and monoclonal antibody preparation. The full-length protein can be used or, alternatively, the invention provides antigenic peptide fragments of the antigen for use as immunogens. An antigenic peptide fragment comprises at least 6 amino acid residues of the amino acid sequence of the full length protein and encompasses an epitope thereof such that an antibody raised against the peptide forms a specific immune complex with the full length protein or with any fragment that contains the epitope. Preferably, the antigenic peptide comprises at least 10 amino acid residues, or at least 15 amino acid residues, or at least 20 amino acid residues, or at least 30 amino acid residues. Preferred epitopes encompassed by the antigenic peptide are regions of the protein that are located on its surface; commonly these are hydrophilic regions.

[0113] In certain embodiments of the invention, at least one epitope encompassed by the antigenic peptide is a region of GPCRX-related protein that is located on the surface of the protein, e.g., a hydrophilic region. A hydrophobicity analysis of the human GPCRX-related protein sequence will indicate which regions of a GPCRX-related protein are particularly hydrophilic and, therefore, are likely to encode surface residues useful for targeting antibody production. As a means for targeting antibody production, hydropathy plots showing regions of hydrophilicity and hydrophobicity may be generated by any method well known in the art, including, for example, the Kyte Doolittle or the Hopp Woods methods, either with or without Fourier transformation. See, e.g., Hopp and Woods, 1981, Proc. Nat. Acad. Sci. USA 78: 3824-3828; Kyte and Doolittle 1982, J. Mol. Biol. 157: 105-142, each of which is incorporated herein by reference in its entirety. Antibodies that are specific for one or more domains within an antigenic protein, or derivatives, fragments, analogs or homologs thereof, are also provided herein.

[0114] A protein of the invention, or a derivative, fragment, analog, homolog or ortholog thereof, may be utilized as an immunogen in the generation of antibodies that immunospecifically bind these protein components.

[0115] Various procedures known within the art may be used for the production of polyclonal or monoclonal antibodies directed against a protein of the invention, or against derivatives, fragments, analogs homologs or orthologs thereof (see, for example, Antibodies: A Laboratory Manual, Harlow and Lane, 1988, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., incorporated herein by reference). Some of these antibodies are discussed below.

[0116] Polyclonal Antibodies

[0117] For the production of polyclonal antibodies, various suitable host animals (e.g., rabbit, goat, mouse or other mammal) may be immunized by one or more injections with the native protein, a synthetic variant thereof, or a derivative of the foregoing. An appropriate immunogenic preparation can contain, for example, the naturally occurring immunogenic protein, a chemically synthesized polypeptide representing the immunogenic protein, or a recombinantly expressed immunogenic protein. Furthermore, the protein may be conjugated to a second protein known to be immunogenic in the mammal being immunized. Examples of such immunogenic proteins include but are not limited to keyhole limpet hemocyanin, serum albumin, bovine thyroglobulin, and soybean trypsin inhibitor. The preparation can further include an adjuvant. Various adjuvants used to increase the immunological response include, but are not limited to, Freund's (complete and incomplete), mineral gels (e.g., aluminum hydroxide), surface active substances (e.g., lysolecithin, pluronic polyols, polyanions, peptides, oil emulsions, dinitrophenol, etc.), adjuvants usable in humans such as Bacille Calmette-Guerin and Corynebacterium parvum, or similar immunostimulatory agents. Additional examples of adjuvants which can be employed include MPL-TDM adjuvant (monophosphoryl Lipid A, synthetic trehalose dicorynomycolate).

[0118] The polyclonal antibody molecules directed against the immunogenic protein can be isolated from the mammal (e.g., from the blood) and further purified by well known techniques, such as affinity chromatography using protein A or protein G, which provide primarily the IgG fraction of immune serum. Subsequently, or alternatively, the specific antigen which is the target of the immunoglobulin sought, or an epitope thereof, may be immobilized on a column to purify the immune specific antibody by immunoaffinity chromatography. Purification of immunoglobulins is discussed, for example, by D. Wilkinson (The Scientist, published by The Scientist, Inc., Philadelphia Pa., Vol. 14, No. 8 (Apr. 17, 2000), pp. 25-28).

[0119] Monoclonal Antibodies

[0120] The term “monoclonal antibody” (MAb) or “monoclonal antibody composition”, as used herein, refers to a population of antibody molecules that contain only one molecular species of antibody molecule consisting of a unique light chain gene product and a unique heavy chain gene product. In particular, the complementarity determining regions (CDRs) of the monoclonal antibody are identical in all the molecules of the population. MAbs thus contain an antigen binding site capable of immunoreacting with a particular epitope of the antigen characterized by a unique binding affinity for it.

[0121] Monoclonal antibodies can be prepared using hybridoma methods, such as those described by Kohler and Milstein, Nature, 256:495 (1975). In a hybridoma method, a mouse, hamster, or other appropriate host animal, is typically immunized with an immunizing agent to elicit lymphocytes that produce or are capable of producing antibodies that will specifically bind to the immunizing agent. Alternatively, the lymphocytes can be immunized in vitro.

[0122] The immunizing agent will typically include the protein antigen, a fragment thereof or a fusion protein thereof. Generally, either peripheral blood lymphocytes are used if cells of human origin are desired, or spleen cells or lymph node cells are used if non-human mammalian sources are desired. The lymphocytes are then fused with an immortalized cell line using a suitable fusing agent, such as polyethylene glycol, to form a hybridoma cell (Goding, MONOCLONAL ANTIBODIES: PRINCIPLES AND PRACTICE, Academic Press, (1986) pp. 59-103). Immortalized cell lines are usually transformed mammalian cells, particularly myeloma cells of rodent, bovine and human origin. Usually, rat or mouse myeloma cell lines are employed. The hybridoma cells can be cultured in a suitable culture medium that preferably contains one or more substances that inhibit the growth or survival of the unfused, immortalized cells. For example, if the parental cells lack the enzyme hypoxanthine guanine phosphoribosyl transferase (HGPRT or HPRT), the culture medium for the hybridomas typically will include hypoxanthine, aminopterin, and thymidine (“HAT medium”), which substances prevent the growth of HGPRT-deficient cells.

[0123] Preferred immortalized cell lines are those that fuse efficiently, support stable high level expression of antibody by the selected antibody-producing cells, and are sensitive to a medium such as HAT medium. More preferred immortalized cell lines are murine myeloma lines, which can be obtained, for instance, from the Salk Institute Cell Distribution Center, San Diego, Calif. and the American Type Culture Collection, Manassas, Va. Human myeloma and mouse-human heteromyeloma cell lines also have been described for the production of human monoclonal antibodies (Kozbor, J. Immunol., 133:3001 (1984); Brodeur et al., MONOCLONAL ANTIBODY PRODUCTION TECHNIQUES AND APPLICATIONS, Marcel Dekker, Inc., New York, (1987) pp. 51-63).

[0124] The culture medium in which the hybridoma cells are cultured can then be assayed for the presence of monoclonal antibodies directed against the antigen. Preferably, the binding specificity of monoclonal antibodies produced by the hybridoma cells is determined by immunoprecipitation or by an in vitro binding assay, such as radioimmunoassay (RIA) or enzyme-linked immunoabsorbent assay (ELISA). Such techniques and assays are known in the art. The binding affinity of the monoclonal antibody can, for example, be determined by the Scatchard analysis of Munson and Pollard, Anal. Biochem., 107:220 (1980). Preferably, antibodies having a high degree of specificity and a high binding affinity for the target antigen are isolated.

[0125] After the desired hybridoma cells are identified, the clones can be subcloned by limiting dilution procedures and grown by standard methods. Suitable culture media for this purpose include, for example, Dulbecco's Modified Eagle's Medium and RPMI-1640 medium. Alternatively, the hybridoma cells can be grown in vivo as ascites in a mammal.

[0126] The monoclonal antibodies secreted by the subclones can be isolated or purified from the culture medium or ascites fluid by conventional immunoglobulin purification procedures such as, for example, protein A-Sepharose, hydroxylapatite chromatography, gel electrophoresis, dialysis, or affinity chromatography.

[0127] The monoclonal antibodies can also be made by recombinant DNA methods, such as those described in U.S. Pat. No. 4,816,567. DNA encoding the monoclonal antibodies of the invention can be readily isolated and sequenced using conventional procedures (e.g., by using oligonucleotide probes that are capable of binding specifically to genes encoding the heavy and light chains of murine antibodies). The hybridoma cells of the invention serve as a preferred source of such DNA. Once isolated, the DNA can be placed into expression vectors, which are then transfected into host cells such as simian COS cells, Chinese hamster ovary (CHO) cells, or myeloma cells that do not otherwise produce immunoglobulin protein, to obtain the synthesis of monoclonal antibodies in the recombinant host cells. The DNA also can be modified, for example, by substituting the coding sequence for human heavy and light chain constant domains in place of the homologous murine sequences (U.S. Pat. No. 4,816,567; Morrison, Nature 368, 812-13 (1994)) or by covalently joining to the immunoglobulin coding sequence all or part of the coding sequence for a non-immunoglobulin polypeptide. Such a non-immunoglobulin polypeptide can be substituted for the constant domains of an antibody of the invention, or can be substituted for the variable domains of one antigen-combining site of an antibody of the invention to create a chimeric bivalent antibody.

[0128] Humanized Antibodies

[0129] The antibodies directed against the protein antigens of the invention can further comprise humanized antibodies or human antibodies. These antibodies are suitable for administration to humans without engendering an immune response by the human against the administered immunoglobulin. Humanized forms of antibodies are chimeric immunoglobulins, immunoglobulin chains or fragments thereof (such as Fv, Fab, Fab′, F(ab′)2 or other antigen-binding subsequences of antibodies) that are principally comprised of the sequence of a human -immunoglobulin, and contain minimal sequence derived from a non-human immunoglobulin. Humanization can be performed following the method of Winter and co-workers (Jones et al., Nature, 321:522-525 (1986); Riechmann et al., Nature, 332:323-327 (1988); Verhoeyen et al., Science, 239:1534-1536 (1988)), by substituting rodent CDRs or CDR sequences for the corresponding sequences of a human antibody. (See also U.S. Pat. No. 5,225,539.) In some instances, Fv framework residues of the human immunoglobulin are replaced by corresponding non-human residues. Humanized antibodies can also comprise residues which are found neither in the recipient antibody nor in the imported CDR or framework sequences. In general, the humanized antibody will comprise substantially all of at least one, and typically two, variable domains, in which all or substantially all of the CDR regions correspond to those of a non-human immunoglobulin and all or substantially all of the framework regions are those of a human immunoglobulin consensus sequence. The humanized antibody optimally also will comprise at least a portion of an immunoglobulin constant region (Fc), typically that of a human immunoglobulin (Jones et al., 1986; Riechmann et al., 1988; and Presta, Curr. Op. Struct. Biol., 2:593-596 (1992)).

[0130] Human Antibodies

[0131] Fully human antibodies relate to antibody molecules in which essentially the entire sequences of both the light chain and the heavy chain, including the CDRs, arise from human genes. Such antibodies are termed “human antibodies”, or “fully human antibodies” herein. Human monoclonal antibodies can be prepared by the trioma technique; the human B-cell hybridoma technique (see Kozbor, et al., 1983 Immunol Today 4: 72) and the EBV hybridoma technique to produce human monoclonal antibodies (see Cole, et al., 1985 In: MONOCLONAL ANTIBODIES AND CANCER THERAPY, Alan R. Liss, Inc., pp. 77-96). Human monoclonal antibodies may be utilized in the practice of the present invention and may be produced by using human hybridomas (see Cote, et al., 1983. Proc Natl Acad Sci USA 80: 2026-2030) or by transforming human B-cells with Epstein Barr Virus in vitro (see Cole, et al., 1985 In: MONOCLONAL ANTIBODIES AND CANCER THERAPY, Alan R. Liss, Inc., pp. 77-96).

[0132] In addition, human antibodies can also be produced using additional techniques, including phage display libraries (Hoogenboom and Winter, J. Mol. Biol., 227:381 (1991); Marks et al., J. Mol. Biol., 222:581 (1991)). Similarly, human antibodies can be made by introducing human immunoglobulin loci into transgenic animals, e.g., mice in which the endogenous immunoglobulin genes have been partially or completely inactivated. Upon challenge, human antibody production is observed, which closely resembles that seen in humans in all respects, including gene rearrangement, assembly, and antibody repertoire. This approach is described, for example, in U.S. Pat. Nos. 5,545,807; 5,545,806; 5,569,825; 5,625,126; 5,633,425; 5,661,016, and in Marks et al. (Bio/Technology 10, 779-783 (1992)); Lonberg et al. (Nature 368 856-859 (1994)); Morrison (Nature 368, 812-13 (1994)); Fishwild et al, (Nature Biotechnology 14, 845-51 (1996)); Neuberger (Nature Biotechnology 14, 826 (1996)); and Lonberg and Huszar (Intern. Rev. Immunol. 13 65-93 (1995)).

[0133] Human antibodies may additionally be produced using transgenic nonhuman animals which are modified so as to produce fully human antibodies rather than the animal's endogenous antibodies in response to challenge by an antigen. (See PCT publication WO94/02602). The endogenous genes encoding the heavy and light immunoglobulin chains in the nonhuman host have been incapacitated, and active loci encoding human heavy and light chain immunoglobulins are inserted into the host's genome. The human genes are incorporated, for example, using yeast artificial chromosomes containing the requisite human DNA segments. An animal which provides all the desired modifications is then obtained as progeny by crossbreeding intermediate transgenic animals containing fewer than the full complement of the modifications. The preferred embodiment of such a nonhuman animal is a mouse, and is termed the Xenomouse™ as disclosed in PCT publications WO 96/33735 and WO 96/34096. This animal produces B cells which secrete fully human immunoglobulins. The antibodies can be obtained directly from the animal after immunization with an immunogen of interest, as, for example, a preparation of a polyclonal antibody, or alternatively from immortalized B cells derived from the animal, such as hybridomas producing monoclonal antibodies. Additionally, the genes encoding the immunoglobulins with human variable regions can be recovered and expressed to obtain the antibodies directly, or can be further modified to obtain analogs of antibodies such as, for example, single chain Fv molecules.

[0134] An example of a method of producing a nonhuman host, exemplified as a mouse, lacking expression of an endogenous immunoglobulin heavy chain is disclosed in U.S. Pat. No. 5,939,598. It can be obtained by a method including deleting the J segment genes from at least one endogenous heavy chain locus in an embryonic stem cell to prevent rearrangement of the locus and to prevent formation of a transcript of a rearranged immunoglobulin heavy chain locus, the deletion being effected by a targeting vector containing a gene encoding a selectable marker; and producing from the embryonic stem cell a transgenic mouse whose somatic and germ cells contain the gene encoding the selectable marker.

[0135] A method for producing an antibody of interest, such as a human antibody, is disclosed in U.S. Pat. No. 5,916,771. It includes introducing an expression vector that contains a nucleotide sequence encoding a heavy chain into one mammalian host cell in culture, introducing an expression vector containing a nucleotide sequence encoding a light chain into another mammalian host cell, and fusing the two cells to form a hybrid cell. The hybrid cell expresses an antibody containing the heavy chain and the light chain.

[0136] In a further improvement on this procedure, a method for identifying a clinically relevant epitope on an immunogen, and a correlative method for selecting an antibody that binds immunospecifically to the relevant epitope with high affinity, are disclosed in PCT publication WO 99/53049.

[0137] Fab Fragments and Single Chain Antibodies

[0138] According to the invention, techniques can be adapted for the production of single-chain antibodies specific to an antigenic protein of the invention (see e.g., U.S. Pat. No. 4,946,778). In addition, methods can be adapted for the construction of Fab expression libraries (see e.g., Huse, et al., 1989 Science 246: 1275-1281) to allow rapid and effective identification of monoclonal Fab fragments with the desired specificity for a protein or derivatives, fragments, analogs or homologs thereof. Antibody fragments that contain the idiotypes to a protein antigen may be produced by techniques known in the art including, but not limited to: (i) an F(ab′)2 fragment produced by pepsin digestion of an antibody molecule; (ii) an Fab fragment generated by reducing the disulfide bridges of an F(ab′)2 fragment; (iii) an Fab fragment generated by the treatment of the antibody molecule with papain and a reducing agent and (iv) Fv fragments.

[0139] Bispecific Antibodies

[0140] Bispecific antibodies are monoclonal, preferably human or humanized, antibodies that have binding specificities for at least two different antigens. In the present case, one of the binding specificities is for an antigenic protein of the invention. The second binding target is any other antigen, and advantageously is a cell-surface protein or receptor or receptor subunit.

[0141] Methods for making bispecific antibodies are known in the art. Traditionally, the recombinant production of bispecific antibodies is based on the co-expression of two immunoglobulin heavy-chain/light-chain pairs, where the two heavy chains have different specificities (Milstein and Cuello, Nature, 305:537-539 (1983)). Because of the random assortment of immunoglobulin heavy and light chains, these hybridomas (quadromas) produce a potential mixture of ten different antibody molecules, of which only one has the correct bispecific structure. The purification of the correct molecule is usually accomplished by affinity chromatography steps. Similar procedures are disclosed in WO 93/08829, published May 13, 1993, and in Traunecker et al., 1991 EMBO J., 10:3655-3659.

[0142] Antibody variable domains with the desired binding specificities (antibody-antigen combining sites) can be fused to immunoglobulin constant domain sequences. The fusion preferably is with an immunoglobulin heavy-chain constant domain, comprising at least part of the hinge, CH2, and CH3 regions. It is preferred to have the first heavy-chain constant region (CH1) containing the site necessary for light-chain binding present in at least one of the fusions. DNAs encoding the immunoglobulin heavy-chain fusions and, if desired, the immunoglobulin light chain, are inserted into separate expression vectors, and are co-transfected into a suitable host organism. For further details of generating bispecific antibodies see, for example, Suresh et al., Methods in Enzymology, 121:210 (1986).

[0143] According to another approach described in WO 96/27011, the interface between a pair of antibody molecules can be engineered to maximize the percentage of heterodimers which are recovered from recombinant cell culture. The preferred interface comprises at least a part of the CH3 region of an antibody constant domain. In this method, one or more small amino acid side chains from the interface of the first antibody molecule are replaced with larger side chains (e.g. tyrosine or tryptophan). Compensatory “cavities” of identical or similar size to the large side chain(s) are created on the interface of the second antibody molecule by replacing large amino acid side chains with smaller ones (e.g. alanine or threonine). This provides a mechanism for increasing the yield of the heterodimer over other unwanted end-products such as homodimers.

[0144] Bispecific antibodies can be prepared as full length antibodies or antibody fragments (e.g. F(ab′)2 bispecific antibodies). Techniques for generating bispecific antibodies from antibody fragments have been described in the literature. For example, bispecific antibodies can be prepared using chemical linkage. Brennan et al., Science 229:81 (1985) describe a procedure wherein intact antibodies are proteolytically cleaved to generate F(ab′)2 fragments. These fragments are reduced in the presence of the dithiol complexing agent sodium arsenite to stabilize vicinal dithiols and prevent intermolecular disulfide formation. The Fab′ fragments generated are then converted to thionitrobenzoate (TNB) derivatives. One of the Fab′-TNB derivatives is then reconverted to the Fab′-thiol by reduction with mercaptoethylamine and is mixed with an equimolar amount of the other Fab′-TNB derivative to form the bispecific antibody. The bispecific antibodies produced can be used as agents for the selective immobilization of enzymes.

[0145] Additionally, Fab′ fragments can be directly recovered from E. coli and chemically coupled to form bispecific antibodies. Shalaby et al., J. Exp. Med. 175:217-225 (1992) describe the production of a fully humanized bispecific antibody F(ab′)2 molecule. Each Fab′ fragment was separately secreted from E. coli and subjected to directed chemical coupling in vitro to form the bispecific antibody. The bispecific antibody thus formed was able to bind to cells overexpressing the ErbB2 receptor and normal human T cells, as well as trigger the lytic activity of human cytotoxic lymphocytes against human breast tumor targets.

[0146] Various techniques for making and isolating bispecific antibody fragments directly from recombinant cell culture have also been described. For example, bispecific antibodies have been produced using leucine zippers. Kostelny et al., J. Immunol. 148(5):1547-1553 (1992). The leucine zipper peptides from the Fos and Jun proteins were linked to the Fab′ portions of two different antibodies by gene fusion. The antibody homodimers were reduced at the hinge region to form monomers and then re-oxidized to form the antibody heterodimers. This method can also be utilized for the production of antibody homodimers. The “diabody” technology described by Hollinger et al., Proc. Natl. Acad. Sci. USA 90:6444-6448 (1993) has provided an alternative mechanism for making bispecific antibody fragments. The fragments comprise a heavy-chain variable domain (VH) connected to a light-chain variable domain (VL) by a linker which is too short to allow pairing between the two domains on the same chain. Accordingly, the VH and VL domains of one fragment are forced to pair with the complementary VL and VH domains of another fragment, thereby forming two antigen-binding sites. Another strategy for making bispecific antibody fragments by the use of single-chain Fv (sFv) dimers has also been reported. See, Gruber et al., J. Immunol. 152:5368 (1994).

[0147] Antibodies with more than two valencies are contemplated. For example, trispecific antibodies can be prepared. Tutt et al., J. Immunol. 147:60 (1991).

[0148] Exemplary bispecific antibodies can bind to two different epitopes, at least one of which originates in the protein antigen of the invention. Alternatively, an anti-antigenic arm of an immunoglobulin molecule can be combined with an arm which binds to a triggering molecule on a leukocyte such as a T-cell receptor molecule (e.g. CD2, CD3, CD28, or B7), or Fc receptors for IgG (FcγR), such as FcγRI (CD64), FcγRII (CD32) and FcγRIII (CD16) so as to focus cellular defense mechanisms to the cell expressing the particular antigen. Bispecific antibodies can also be used to direct cytotoxic agents to cells which express a particular antigen. These antibodies possess an antigen-binding arm and an arm which binds a cytotoxic agent or a radionuclide chelator, such as EOTUBE, DPTA, DOTA, or TETA. Another bispecific antibody of interest binds the protein antigen described herein and further binds tissue factor (TF).

[0149] Heteroconjugate Antibodies

[0150] Heteroconjugate antibodies are also within the scope of the present invention. Heteroconjugate antibodies are composed of two covalently joined antibodies. Such antibodies have, for example, been proposed to target immune system cells to unwanted cells (U.S. Pat. No. 4,676,980), and for treatment of HIV infection (WO 91/00360; WO 92/200373; EP 03089). It is contemplated that the antibodies can be prepared in vitro using known methods in synthetic protein chemistry, including those involving crosslinking agents. For example, immunotoxins can be constructed using a disulfide exchange reaction or by forming a thioether bond. Examples of suitable reagents for this purpose include iminothiolate and methyl-4-mercaptobutyrimidate and those disclosed, for example, in U.S. Pat. No. 4,676,980.

[0151] Effector Function Engineering

[0152] It can be desirable to modify the antibody of the invention with respect to effector function, so as to enhance, e.g., the effectiveness of the antibody in treating cancer. For example, cysteine residue(s) can be introduced into the Fc region, thereby allowing interchain disulfide bond formation in this region. The homodimeric antibody thus generated can have improved internalization capability and/or increased complement-mediated cell killing and antibody-dependent cellular cytotoxicity (ADCC). See Caron et al., J. Exp Med., 176: 1191-1195 (1992) and Shopes, J. Immunol., 148: 2918-2922 (1992). Homodimeric antibodies with enhanced anti-tumor activity can also be prepared using heterobifunctional cross-linkers as described in Wolff et al. Cancer Research, 53: 2560-2565 (1993). Alternatively, an antibody can be engineered that has dual Fc regions and can thereby have enhanced complement lysis and ADCC capabilities. See Stevenson et al., Anti-Cancer Drug Design, 3: 219-230 (1989).

[0153] Immunoconjugates

[0154] The invention also pertains to immunoconjugates comprising an antibody conjugated to a cytotoxic agent such as a chemotherapeutic agent, toxin (e.g., an enzymatically active toxin of bacterial, fungal, plant, or animal origin, or fragments thereof), or a radioactive isotope (i.e., a radioconjugate).

[0155] Chemotherapeutic agents useful in the generation of such immunoconjugates have been described above. Enzymatically active toxins and fragments thereof that can be used include diphtheria A chain, nonbinding active fragments of diphtheria toxin, exotoxin A chain (from Pseudomonas aeruginosa), ricin A chain, abrin A chain, modeccin A chain, alpha-sarcin, Aleurites fordii proteins, dianthin proteins, Phytolaca americana proteins (PAPI, PAPII, and PAP-S), momordica charantia inhibitor, curcin, crotin, sapaonaria officinalis inhibitor, gelonin, mitogellin, restrictocin, phenomycin, enomycin, and the tricothecenes. A variety of radionuclides are available for the production of radioconjugated antibodies. Examples include 212Bi, 131I, 131In, 90Y, and 186Re.

[0156] Conjugates of the antibody and cytotoxic agent are made using a variety of bifunctional protein-coupling agents such as N-succinimidyl-3-(2-pyridyldithiol) propionate (SPDP), iminothiolane (IT), bifunctional derivatives of imidoesters (such as dimethyl adipimidate HCL), active esters (such as disuccinimidyl suberate), aldehydes (such as glutareldehyde), bis-azido compounds (such as bis (p-azidobenzoyl) hexanediamine), bis-diazonium derivatives (such as bis-(p-diazoniumbenzoyl)-ethylenediamine), diisocyanates (such as tolyene 2,6-diisocyanate), and bis-active fluorine compounds (such as 1,5-difluoro-2,4-dinitrobenzene). For example, a ricin immunotoxin can be prepared as described in Vitetta et al., Science, 238: 1098 (1987). Carbon-14-labeled 1-isothiocyanatobenzyl-3-methyldiethylene triaminepentaacetic acid (MX-DTPA) is an exemplary chelating agent for conjugation of radionucleotide to the antibody. See WO94/11026.

[0157] In another embodiment, the antibody can be conjugated to a “receptor” (such streptavidin) for utilization in tumor pretargeting wherein the antibody-receptor conjugate is administered to the patient, followed by removal of unbound conjugate from the circulation using a clearing agent and then administration of a “ligand” (e.g., avidin) that is in turn conjugated to a cytotoxic agent.

[0158] In one embodiment, methods for the screening of antibodies that possess the desired specificity include, but are not limited to, enzyme-linked immunosorbent assay (ELISA) and other immunologically-mediated techniques known within the art. In a specific embodiment, selection of antibodies that are specific to a particular domain of an GPCRX protein is facilitated by generation of hybridomas that bind to the fragment of an GPCRX protein possessing such a domain. Thus, antibodies that are specific for a desired domain within an GPCRX protein, or derivatives, fragments, analogs or homologs thereof, are also provided herein.

[0159] Anti-GPCRX antibodies may be used in methods known within the art relating to the localization and/or quantitation of an GPCRX protein (e.g., for use in measuring levels of the GPCRX protein within appropriate physiological samples, for use in diagnostic methods, for use in imaging the protein, and the like). In a given embodiment, antibodies for GPCRX proteins, or derivatives, fragments, analogs or homologs thereof, that contain the antibody derived binding domain, are utilized as pharmacologically-active compounds (hereinafter “Therapeutics”).

[0160] An anti-GPCRX antibody (e.g., monoclonal antibody) can be used to isolate an GPCRX polypeptide by standard techniques, such as affinity chromatography or immunoprecipitation. An anti-GPCRX antibody can facilitate the purification of natural GPCRX polypeptide from cells and of recombinantly-produced GPCRX polypeptide expressed in host cells. Moreover, an anti-GPCRX antibody can be used to detect GPCRX protein (e.g., in a cellular lysate or cell supernatant) in order to evaluate the abundance and pattern of expression of the GPCRX protein. Anti-GPCRX antibodies can be used diagnostically to monitor protein levels in tissue as part of a clinical testing procedure, e.g. to, for example, determine the efficacy of a given treatment regimen. Detection can be facilitated by coupling (i.e., physically linking) the antibody to a detectable substance. Examples of detectable substances include various enzymes, prosthetic groups, fluorescent materials, luminescent materials, bioluminescent materials, and radioactive materials. Examples of suitable enzymes include horseradish peroxidase, alkaline phosphatase, β-galactosidase, or acetylcholinesterase; examples of suitable prosthetic group complexes include streptavidin/biotin and avidin/biotin; examples of suitable fluorescent materials include umbelliferone, fluorescein, fluorescein isothiocyanate, rhodamine, dichlorotriazinylamine fluorescein, dansyl chloride or phycoerythrin; an example of a luminescent material includes luminol; examples of bioluminescent materials include luciferase, luciferin, and aequorin, and examples of suitable radioactive material include 125I, 131I, 35S or 3H.

[0161] GPCRX Recombinant Expression Vectors and Host Cells

[0162] Another aspect of the invention pertains to vectors, preferably expression vectors, containing a nucleic acid encoding an GPCRX protein, or derivatives, fragments, analogs or homologs thereof. As used herein, the term “vector” refers to a nucleic acid molecule capable of transporting another nucleic acid to which it has been linked. One type of vector is a “plasmid”, which refers to a circular double stranded DNA loop into which additional DNA segments can be ligated. Another type of vector is a viral vector, wherein additional DNA segments can be ligated into the viral genome. Certain vectors are capable of autonomous replication in a host cell into which they are introduced (e.g., bacterial vectors having a bacterial origin of replication and episomal mammalian vectors). Other vectors (e.g., non-episomal mammalian vectors) are integrated into the genome of a host cell upon introduction into the host cell, and thereby are replicated along with the host genome. Moreover, certain vectors are capable of directing the expression of genes to which they are operatively-linked. Such vectors are referred to herein as “expression vectors”. In general, expression vectors of utility in recombinant DNA techniques are often in the form of plasmids. In the present specification, “plasmid” and “vector” can be used interchangeably as the plasmid is the most commonly used form of vector. However, the invention is intended to include such other forms of expression vectors, such as viral vectors (e.g., replication defective retroviruses, adenoviruses and adeno-associated viruses), which serve equivalent functions.

[0163] The recombinant expression vectors of the invention comprise a nucleic acid of the invention in a form suitable for expression of the nucleic acid in a host cell, which means that the recombinant expression vectors include one or more regulatory sequences, selected on the basis of the host cells to be used for expression, that is operatively-linked to the nucleic acid sequence to be expressed. Within a recombinant expression vector, “operably-linked” is intended to mean that the nucleotide sequence of interest is linked to the regulatory sequence(s) in a manner that allows for expression of the nucleotide sequence (e.g., in an in vitro transcription/translation system or in a host cell when the vector is introduced into the host cell).

[0164] The term “regulatory sequence” is intended to includes promoters, enhancers and other expression control elements (e.g., polyadenylation signals). Such regulatory sequences are described, for example, in Goeddel, GENE EXPRESSION TECHNOLOGY: METHODS IN ENZYMOLOGY 185, Academic Press, San Diego, Calif. (1990). Regulatory sequences include those that direct constitutive expression of a nucleotide sequence in many types of host cell and those that direct expression of the nucleotide sequence only in certain host cells (e.g., tissue-specific regulatory sequences). It will be appreciated by those skilled in the art that the design of the expression vector can depend on such factors as the choice of the host cell to be transformed, the level of expression of protein desired, etc. The expression vectors of the invention can be introduced into host cells to thereby produce proteins or peptides, including fusion proteins or peptides, encoded by nucleic acids as described herein (e.g., GPCRX proteins, mutant forms of GPCRX proteins, fusion proteins, etc.).

[0165] The recombinant expression vectors of the invention can be designed for expression of GPCRX proteins in prokaryotic or eukaryotic cells. For example, GPCRX proteins can be expressed in bacterial cells such as Escherichia coli, insect cells (using baculovirus expression vectors) yeast cells or mammalian cells. Suitable host cells are discussed further in Goeddel, GENE EXPRESSION TECHNOLOGY: METHODS IN ENZYMOLOGY 185, Academic Press, San Diego, Calif. (1990). Alternatively, the recombinant expression vector can be transcribed and translated in vitro, for example using T7 promoter regulatory sequences and T7 polymerase.

[0166] Expression of proteins in prokaryotes is most often carried out in Escherichia coli with vectors containing constitutive or inducible promoters directing the expression of either fusion or non-fusion proteins. Fusion vectors add a number of amino acids to a protein encoded therein, usually to the amino terminus of the recombinant protein. Such fusion vectors typically serve three purposes: (i) to increase expression of recombinant protein; (ii) to increase the solubility of the recombinant protein; and (iii) to aid in the purification of the recombinant protein by acting as a ligand in affinity purification. Often, in fusion expression vectors, a proteolytic cleavage site is introduced at the junction of the fusion moiety and the recombinant protein to enable separation of the recombinant protein from the fusion moiety subsequent to purification of the fusion protein. Such enzymes, and their cognate recognition sequences, include Factor Xa, thrombin and enterokinase. Typical fusion expression vectors include pGEX (Pharmacia Biotech Inc; Smith and Johnson, 1988. Gene 67: 31-40), pMAL (New England Biolabs, Beverly, Mass.) and pRIT5 (Pharmacia, Piscataway, N.J.) that fuse glutathione S-transferase (GST), maltose E binding protein, or protein A, respectively, to the target recombinant protein.

[0167] Examples of suitable inducible non-fusion E. coli expression vectors include pTrc (Amrann et al., (1988) Gene 69:301-315) and pET 11d (Studier et al., GENE EXPRESSION TECHNOLOGY: METHODS IN ENZYMOLOGY 185, Academic Press, San Diego, Calif. (1990) 60-89).

[0168] One strategy to maximize recombinant protein expression in E. coli is to express the protein in a host bacteria with an impaired capacity to proteolytically cleave the recombinant protein. See, e.g., Gottesman, GENE EXPRESSION TECHNOLOGY: METHODS IN ENZYMOLOGY 185, Academic Press, San Diego, Calif. (1990) 119-128. Another strategy is to alter the nucleic acid sequence of the nucleic acid to be inserted into an expression vector so that the individual codons for each amino acid are those preferentially utilized in E. coli (see, e.g., Wada, et al., 1992. Nucl. Acids Res. 20: 2111-2118). Such alteration of nucleic acid sequences of the invention can be carried out by standard DNA synthesis techniques.

[0169] In another embodiment, the GPCRX expression vector is a yeast expression vector. Examples of vectors for expression in yeast Saccharomyces cerivisae include pYepSec1 (Baldari, et al., 1987. EMBO J. 6: 229-234), pMFa (Kudjan and Herskowitz, 1982. Cell 30: 933-943), pJRY88 (Schultz et al., 1987. Gene 54: 113-123), pYES2 (Invitrogen Corporation, San Diego, Calif.), and picZ (InVitrogen Corp, San Diego, Calif.).

[0170] Alternatively, GPCRX can be expressed in insect cells using baculovirus expression vectors. Baculovirus vectors available for expression of proteins in cultured insect cells (e.g., SF9 cells) include the pAc series (Smith, et al., 1983. Mol. Cell. Biol. 3: 2156-2165) and the pVL series (Lucklow and Summers, 1989. Virology 170: 31-39).

[0171] In yet another embodiment, a nucleic acid of the invention is expressed in mammalian cells using a mammalian expression vector. Examples of mammalian expression vectors include pCDM8 (Seed, 1987. Nature 329: 840) and pMT2PC (Kaufman, et al., 1987. EMBO J. 6: 187-195). When used in mammalian cells, the expression vector's control functions are often provided by viral regulatory elements. For example, commonly used promoters are derived from polyoma, adenovirus 2, cytomegalovirus, and simian virus 40. For other suitable expression systems for both prokaryotic and eukaryotic cells see, e.g., Chapters 16 and 17 of Sambrook, et al., MOLECULAR CLONING: A LABORATORY MANUAL. 2nd ed., Cold Spring Harbor Laboratory, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., 1989.

[0172] In another embodiment, the recombinant mammalian expression vector is capable of directing expression of the nucleic acid preferentially in a particular cell type (e.g., tissue-specific regulatory elements are used to express the nucleic acid). Tissue-specific regulatory elements are known in the art. Non-limiting examples of suitable tissue-specific promoters include the albumin promoter (liver-specific; Pinkert, et al., 1987. Genes Dev. 1: 268-277), lymphoid-specific promoters (Calame and Eaton, 1988. Adv. Immunol. 43: 235-275), in particular promoters of T cell receptors (Winoto and Baltimore, 1989. EMBO J. 8: 729-733) and immunoglobulins (Baneiji, et al., 1983. Cell 33: 729-740; Queen and Baltimore, 1983. Cell 33: 741-748), neuron-specific promoters (e.g. the neurofilament promoter; Byrne and Ruddle, 1989. Proc. Natl. Acad. Sci. USA 86: 5473-5477), pancreas-specific promoters (Edlund, et al., 1985. Science 230: 912-916), and mammary gland-specific promoters (e.g., milk whey promoter; U.S. Pat. No. 4,873,316 and European Application Publication No. 264,166). Developmentally-regulated promoters are also encompassed, e.g., the murine hox promoters (Kessel and Gruss, 1990. Science 249: 374-379) and the α-fetoprotein promoter (Campes and Tilghman, 1989. Genes Dev. 3: 537-546).

[0173] The invention further provides a recombinant expression vector comprising a DNA molecule of the invention cloned into the expression vector in an antisense orientation. That is, the DNA molecule is operatively-linked to a regulatory sequence in a manner that allows for expression (by transcription of the DNA molecule) of an RNA molecule that is antisense to GPCRX mRNA. Regulatory sequences operatively linked to a nucleic acid cloned in the antisense orientation can be chosen that direct the continuous expression of the antisense RNA molecule in a variety of cell types, for instance viral promoters and/or enhancers, or regulatory sequences can be chosen that direct constitutive, tissue specific or cell type specific expression of antisense RNA. The antisense expression vector can be in the form of a recombinant plasmid, phagemid or attenuated virus in which antisense nucleic acids are produced under the control of a high efficiency regulatory region, the activity of which can be determined by the cell type into which the vector is introduced. For a discussion of the regulation of gene expression using antisense genes see, e.g., Weintraub, et al., “Antisense RNA as a molecular tool for genetic analysis,” Reviews-Trends in Genetics, Vol. 1(1) 1986.

[0174] Another aspect of the invention pertains to host cells into which a recombinant expression vector of the invention has been introduced. The terms “host cell” and “recombinant host cell” are used interchangeably herein. It is understood that such terms refer not only to the particular subject cell but also to the progeny or potential progeny of such a cell. Because certain modifications may occur in succeeding generations due to either mutation or environmental influences, such progeny may not, in fact, be identical to the parent cell, but are still included within the scope of the term as used herein.

[0175] A host cell can be any prokaryotic or eukaryotic cell. For example, GPCRX protein can be expressed in bacterial cells such as E. coli, insect cells, yeast or mammalian cells (such as Chinese hamster ovary cells (CHO) or COS cells). Other suitable host cells are known to those skilled in the art.

[0176] Vector DNA can be introduced into prokaryotic or eukaryotic cells via conventional transformation or transfection techniques. As used herein, the terms “transformation” and “transfection” are intended to refer to a variety of art-recognized techniques for introducing foreign nucleic acid (e.g., DNA) into a host cell, including calcium phosphate or calcium chloride co-precipitation, DEAE-dextran-mediated transfection, lipofection, or electroporation. Suitable methods for transforming or transfecting host cells can be found in Sambrook, et al. (MOLECULAR CLONING: A LABORATORY MANUAL. 2nd ed., Cold Spring Harbor Laboratory, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., 1989), and other laboratory manuals.

[0177] For stable transfection of mammalian cells, it is known that, depending upon the expression vector and transfection technique used, only a small fraction of cells may integrate the foreign DNA into their genome. In order to identify and select these integrants, a gene that encodes a selectable marker (e.g., resistance to antibiotics) is generally introduced into the host cells along with the gene of interest. Various selectable markers include those that confer resistance to drugs, such as G418, hygromycin and methotrexate. Nucleic acid encoding a selectable marker can be introduced into a host cell on the same vector as that encoding GPCRX or can be introduced on a separate vector. Cells stably transfected with the introduced nucleic acid can be identified by drug selection (e.g., cells that have incorporated the selectable marker gene will survive, while the other cells die).

[0178] A host cell of the invention, such as a prokaryotic or eukaryotic host cell in culture, can be used to produce (i.e., express) GPCRX protein. Accordingly, the invention further provides methods for producing GPCRX protein using the host cells of the invention. In one embodiment, the method comprises culturing the host cell of invention (into which a recombinant expression vector encoding GPCRX protein has been introduced) in a suitable medium such that GPCRX protein is produced. In another embodiment, the method further comprises isolating GPCRX protein from the medium or the host cell.

[0179] Transgenic GPCRX Animals

[0180] The host cells of the invention can also be used to produce non-human transgenic animals. For example, in one embodiment, a host cell of the invention is a fertilized oocyte or an embryonic stem cell into which GPCRX protein-coding sequences have been introduced. Such host cells can then be used to create non-human transgenic animals in which exogenous GPCRX sequences have been introduced into their genome or homologous recombinant animals in which endogenous GPCRX sequences have been altered. Such animals are useful for studying the function and/or activity of GPCRX protein and for identifying and/or evaluating modulators of GPCRX protein activity. As used herein, a “transgenic animal” is a non-human animal, preferably a mammal, more preferably a rodent such as a rat or mouse, in which one or more of the cells of the animal includes a transgene. Other examples of transgenic animals include non-human primates, sheep, dogs, cows, goats, chickens, amphibians, etc. A transgene is exogenous DNA that is integrated into the genome of a cell from which a transgenic animal develops and that remains in the genome of the mature animal, thereby directing the expression of an encoded gene product in one or more cell types or tissues of the transgenic animal. As used herein, a “homologous recombinant animal” is a non-human animal, preferably a mammal, more preferably a mouse, in which an endogenous GPCRX gene has been altered by homologous recombination between the endogenous gene and an exogenous DNA molecule introduced into a cell of the animal, e.g., an embryonic cell of the animal, prior to development of the animal.

[0181] A transgenic animal of the invention can be created by introducing GPCRX-encoding nucleic acid into the male pronuclei of a fertilized oocyte (e.g., by microinjection, retroviral infection) and allowing the oocyte to develop in a pseudopregnant female foster animal. The human GPCRX cDNA sequences of SEQ ID NOS:1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, 75, 77, 79, 81, 83, 85, 87, 89, 91, 93, 95, 97, 99, 101, 103, 105, 107, 109, 111, 113, 115, 117, 119, 121, 123, 125, 127, 129, 131, 133, 135, 137, 139, 141, 143, 145, 147, 149 and 151 can be introduced as a transgene into the genome of a non-human animal. Alternatively, a non-human homologue of the human GPCRX gene, such as a mouse GPCRX gene, can be isolated based on hybridization to the human GPCRX cDNA (described further supra) and used as a transgene. Intronic sequences and polyadenylation signals can also be included in the transgene to increase the efficiency of expression of the transgene. A tissue-specific regulatory sequence(s) can be operably-linked to the GPCRX transgene to direct expression of GPCRX protein to particular cells. Methods for generating transgenic animals via embryo manipulation and microinjection, particularly animals such as mice, have become conventional in the art and are described, for example, in U.S. Pat. Nos. 4,736,866; 4,870,009; and 4,873,191; and Hogan, 1986. In: MANIPULATING THE MOUSE EMBRYO, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y. Similar methods are used for production of other transgenic animals. A transgenic founder animal can be identified based upon the presence of the GPCRX transgene in its genome and/or expression of GPCRX mRNA in tissues or cells of the animals. A transgenic founder animal can then be used to breed additional animals carrying the transgene. Moreover, transgenic animals carrying a transgene-encoding GPCRX protein can further be bred to other transgenic animals carrying other transgenes.

[0182] To create a homologous recombinant animal, a vector is prepared which contains at least a portion of an GPCRX gene into which a deletion, addition or substitution has been introduced to thereby alter, e.g., functionally disrupt, the GPCRX gene. The GPCRX gene can be a human gene (e.g., the cDNA of SEQ ID NOS:1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, 75, 77, 79, 81, 83, 85, 87, 89, 91, 93, 95, 97, 99, 101, 103, 105, 107, 109, 111, 113, 115, 117, 119, 121, 123, 125, 127, 129, 131, 133, 135, 137, 139, 141, 143, 145, 147, 149 and 151), but more preferably, is a non-human homologue of a human GPCRX gene. For example, a mouse homologue of human GPCRX gene of SEQ ID NOS:1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, 75, 77, 79, 81, 83, 85, 87, 89, 91, 93, 95, 97, 99, 101, 103, 105, 107, 109, 111, 113, 115, 117, 119, 121, 123, 125, 127, 129, 131, 133, 135, 137, 139, 141, 143, 145, 147, 149 and 151 can be used to construct a homologous recombination vector suitable for altering an endogenous GPCRX gene in the mouse genome. In one embodiment, the vector is designed such that, upon homologous recombination, the endogenous GPCRX gene is functionally disrupted (i.e., no longer encodes a functional protein; also referred to as a “knock out” vector).

[0183] Alternatively, the vector can be designed such that, upon homologous recombination, the endogenous GPCRX gene is mutated or otherwise altered but still encodes functional protein (e.g., the upstream regulatory region can be altered to thereby alter the expression of the endogenous GPCRX protein). In the homologous recombination vector, the altered portion of the GPCRX gene is flanked at its 5′- and 3′-termini by additional nucleic acid of the GPCRX gene to allow for homologous recombination to occur between the exogenous GPCRX gene carried by the vector and an endogenous GPCRX gene in an embryonic stem cell. The additional flanking GPCRX nucleic acid is of sufficient length for successful homologous recombination with the endogenous gene. Typically, several kilobases of flanking DNA (both at the 5′- and 3′-termini) are included in the vector. See, e.g., Thomas, et al., 1987. Cell 51: 503 for a description of homologous recombination vectors. The vector is ten introduced into an embryonic stem cell line (e.g., by electroporation) and cells in which the introduced GPCRX gene has homologously-recombined with the endogenous GPCRX gene are selected. See, e.g., Li, et al., 1992. Cell 69: 915.

[0184] The selected cells are then injected into a blastocyst of an animal (e.g., a mouse) to form aggregation chimeras. See, e.g., Bradley, 1987. In: TERATOCARCINOMAS AND EMBRYONIC STEM CELLS: A PRACTICAL APPROACH, Robertson, ed. IRL, Oxford, pp. 113-152. A chimeric embryo can then be implanted into a suitable pseudopregnant female foster animal and the embryo brought to term. Progeny harboring the homologously-recombined DNA in their germ cells can be used to breed animals in which all cells of the animal contain the homologously-recombined DNA by germline transmission of the transgene. Methods for constructing homologous recombination vectors and homologous recombinant animals are described further in Bradley, 1991. Curr. Opin. Biotechnol. 2: 823-829; PCT International Publication Nos.: WO 90/11354; WO 91/01140; WO 92/0968; and WO 93/04169.

[0185] In another embodiment, transgenic non-humans animals can be produced that contain selected systems that allow for regulated expression of the transgene. One example of such a system is the cre/loxP recombinase system of bacteriophage P1. For a description of the cre/loxP recombinase system, See, e.g., Lakso, et al., 1992. Proc. Natl. Acad. Sci. USA 89: 6232-6236. Another example of a recombinase system is the FLP recombinase system of Saccharomyces cerevisiae. See, O'Gorman, et al., 1991. Science 251:1351-1355. If a cre/loxP recombinase system is used to regulate expression of the transgene, animals containing transgenes encoding both the Cre recombinase and a selected protein are required. Such animals can be provided through the construction of “double” transgenic animals, e.g., by mating two transgenic animals, one containing a transgene encoding a selected protein and the other containing a transgene encoding a recombinase.

[0186] Clones of the non-human transgenic animals described herein can also be produced according to the methods described in Wilmut, et al., 1997. Nature 385: 810-813. In brief, a cell (e.g., a somatic cell) from the transgenic animal can be isolated and induced to exit the growth cycle and enter G0 phase. The quiescent cell can then be fused, e.g., through the use of electrical pulses, to an enucleated oocyte from an animal of the same species from which the quiescent cell is isolated. The reconstructed oocyte is then cultured such that it develops to morula or blastocyte and then transferred to pseudopregnant female foster animal. The offspring borne of this female foster animal will be a clone of the animal from which the cell (e.g., the somatic cell) is isolated.

[0187] Pharmaceutical Compositions

[0188] The GPCRX nucleic acid molecules, GPCRX proteins, and anti-GPCRX antibodies (also referred to herein as “active compounds”) of the invention, and derivatives, fragments, analogs and homologs thereof, can be incorporated into pharmaceutical compositions suitable for administration. Such compositions typically comprise the nucleic acid molecule, protein, or antibody and a pharmaceutically acceptable carrier. As used herein, “pharmaceutically acceptable carrier” is intended to include any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like, compatible with pharmaceutical administration. Suitable carriers are described in the most recent edition of Remington's Pharmaceutical Sciences, a standard reference text in the field, which is incorporated herein by reference. Preferred examples of such carriers or diluents include, but are not limited to, water, saline, finger's solutions, dextrose solution, and 5% human serum albumin. Liposomes and non-aqueous vehicles such as fixed oils may also be used. The use of such media and agents for pharmaceutically active substances is well known in the art. Except insofar as any conventional media or agent is incompatible with the active compound, use thereof in the compositions is contemplated. Supplementary active compounds can also be incorporated into the compositions.

[0189] A pharmaceutical composition of the invention is formulated to be compatible with its intended route of administration. Examples of routes of administration include parenteral, e.g., intravenous, intradermal, subcutaneous, oral (e.g., inhalation), transdermal (i.e., topical), transmucosal, and rectal administration. Solutions or suspensions used for parenteral, intradermal, or subcutaneous application can include the following components: a sterile diluent such as water for injection, saline solution, fixed oils, polyethylene glycols, glycerine, propylene glycol or other synthetic solvents; antibacterial agents such as benzyl alcohol or methyl parabens; antioxidants such as ascorbic acid or sodium bisulfite; chelating agents such as ethylenediaminetetraacetic acid (EDTA); buffers such as acetates, citrates or phosphates, and agents for the adjustment of tonicity such as sodium chloride or dextrose. The pH can be adjusted with acids or bases, such as hydrochloric acid or sodium hydroxide. The parenteral preparation can be enclosed in ampoules, disposable syringes or multiple dose vials made of glass or plastic.

[0190] Pharmaceutical compositions suitable for injectable use include sterile aqueous solutions (where water soluble) or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersion. For intravenous administration, suitable carriers include physiological saline, bacteriostatic water, Cremophor EL™ (BASF, Parsippany, N.J.) or phosphate buffered saline (PBS). In all cases, the composition must be sterile and should be fluid to the extent that easy syringeability exists. It must be stable under the conditions of manufacture and storage and must be preserved against the contaminating action of microorganisms such as bacteria and fungi. The carrier can be a solvent or dispersion medium containing, for example, water, ethanol, polyol (for example, glycerol, propylene glycol, and liquid polyethylene glycol, and the like), and suitable mixtures thereof. The proper fluidity can be maintained, for example, by the use of a coating such as lecithin, by the maintenance of the required particle size in the case of dispersion and by the use of surfactants. Prevention of the action of microorganisms can be achieved by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, ascorbic acid, thimerosal, and the like. In many cases, it will be preferable to include isotonic agents, for example, sugars, polyalcohols such as manitol, sorbitol, sodium chloride in the composition. Prolonged absorption of the injectable compositions can be brought about by including in the composition an agent which delays absorption, for example, aluminum monostearate and gelatin.

[0191] Sterile injectable solutions can be prepared by incorporating the active compound (e.g., an GPCRX protein or anti-GPCRX antibody) in the required amount in an appropriate solvent with one or a combination of ingredients enumerated above, as required, followed by filtered sterilization. Generally, dispersions are prepared by incorporating the active compound into a sterile vehicle that contains a basic dispersion medium and the required other ingredients from those enumerated above. In the case of sterile powders for the preparation of sterile injectable solutions, methods of preparation are vacuum drying and freeze-drying that yields a powder of the active ingredient plus any additional desired ingredient from a previously sterile-filtered solution thereof.

[0192] Oral compositions generally include an inert diluent or an edible carrier. They can be enclosed in gelatin capsules or compressed into tablets. For the purpose of oral therapeutic administration, the active compound can be incorporated with excipients and used in the form of tablets, troches, or capsules. Oral compositions can also be prepared using a fluid carrier for use as a mouthwash, wherein the compound in the fluid carrier is applied orally and swished and expectorated or swallowed. Pharmaceutically compatible binding agents, and/or adjuvant materials can be included as part of the composition. The tablets, pills, capsules, troches and the like can contain any of the following ingredients, or compounds of a similar nature: a binder such as microcrystalline cellulose, gum tragacanth or gelatin; an excipient such as starch or lactose, a disintegrating agent such as alginic acid, Primogel, or corn starch; a lubricant such as magnesium stearate or Sterotes; a glidant such as colloidal silicon dioxide; a sweetening agent such as sucrose or saccharin; or a flavoring agent such as peppermint, methyl salicylate, or orange flavoring.

[0193] For administration by inhalation, the compounds are delivered in the form of an aerosol spray from pressured container or dispenser which contains a suitable propellant, e.g., a gas such as carbon dioxide, or a nebulizer.

[0194] Systemic administration can also be by transmucosal or transdermal means. For transmucosal or transdermal administration, penetrants appropriate to the barrier to be permeated are used in the formulation. Such penetrants are generally known in the art, and include, for example, for transmucosal administration, detergents, bile salts, and fusidic acid derivatives. Transmucosal administration can be accomplished through the use of nasal sprays or suppositories. For transdermal administration, the active compounds are formulated into ointments, salves, gels, or creams as generally known in the art.

[0195] The compounds can also be prepared in the form of suppositories (e.g., with conventional suppository bases such as cocoa butter and other glycerides) or retention enemas for rectal delivery.

[0196] In one embodiment, the active compounds are prepared with carriers that will protect the compound against rapid elimination from the body, such as a controlled release formulation, including implants and microencapsulated delivery systems. Biodegradable, biocompatible polymers can be used, such as ethylene vinyl acetate, polyanhydrides, polyglycolic acid, collagen, polyorthoesters, and polylactic acid. Methods for preparation of such formulations will be apparent to those skilled in the art. The materials can also be obtained commercially from Alza Corporation and Nova Pharmaceuticals, Inc. Liposomal suspensions (including liposomes targeted to infected cells with monoclonal antibodies to viral antigens) can also be used as pharmaceutically acceptable carriers. These can be prepared according to methods known to those skilled in the art, for example, as described in U.S. Pat. No. 4,522,811.

[0197] It is especially advantageous to formulate oral or parenteral compositions in dosage unit form for ease of administration and uniformity of dosage. Dosage unit form as used herein refers to physically discrete units suited as unitary dosages for the subject to be treated; each unit containing a predetermined quantity of active compound calculated to produce the desired therapeutic effect in association with the required pharmaceutical carrier. The specification for the dosage unit forms of the invention are dictated by and directly dependent on the unique characteristics of the active compound and the particular therapeutic effect to be achieved, and the limitations inherent in the art of compounding such an active compound for the treatment of individuals.

[0198] The nucleic acid molecules of the invention can be inserted into vectors and used as gene therapy vectors. Gene therapy vectors can be delivered to a subject by, for example, intravenous injection, local administration (see, e.g., U.S. Pat. No. 5,328,470) or by stereotactic injection (see, e.g., Chen, et al., 1994. Proc. Natl. Acad. Sci. USA 91: 3054-3057). The pharmaceutical preparation of the gene therapy vector can include the gene therapy vector in an acceptable diluent, or can comprise a slow release matrix in which the gene delivery vehicle is imbedded. Alternatively, where the complete gene delivery vector can be produced intact from recombinant cells, e.g., retroviral vectors, the pharmaceutical preparation can include one or more cells that produce the gene delivery system.

[0199] The pharmaceutical compositions can be included in a container, pack, or dispenser together with instructions for administration.

[0200] Screening and Detection Methods

[0201] The isolated nucleic acid molecules of the invention can be used to express GPCRX protein (e.g., via a recombinant expression vector in a host cell in gene therapy applications), to detect GPCRX mRNA (e.g., in a biological sample) or a genetic lesion in an GPCRX gene, and to modulate GPCRX activity, as described further, below. In addition, the GPCRX proteins can be used to screen drugs or compounds that modulate the GPCRX protein activity or expression as well as to treat disorders characterized by insufficient or excessive production of GPCRX protein or production of GPCRX protein forms that have decreased or aberrant activity compared to GPCRX wild-type protein (e.g.; diabetes (regulates insulin release); obesity (binds and transport lipids); metabolic disturbances associated with obesity, the metabolic syndrome X as well as anorexia and wasting disorders associated with chronic diseases and various cancers, and infectious disease(possesses anti-microbial activity) and the various dyslipidemias. In addition, the anti-GPCRX antibodies of the invention can be used to detect and isolate GPCRX proteins and modulate GPCRX activity. In yet a further aspect, the invention can be used in methods to influence appetite, absorption of nutrients and the disposition of metabolic substrates in both a positive and negative fashion.

[0202] The invention further pertains to novel agents identified by the screening assays described herein and uses thereof for treatments as described, supra.

[0203] Screening Assays

[0204] The invention provides a method (also referred to herein as a “screening assay”) for identifying modulators, i.e., candidate or test compounds or agents (e.g., peptides, peptidomimetics, small molecules or other drugs) that bind to GPCRX proteins or have a stimulatory or inhibitory effect on, e.g., GPCRX protein expression or GPCRX protein activity. The invention also includes compounds identified in the screening assays described herein.

[0205] In one embodiment, the invention provides assays for screening candidate or test compounds which bind to or modulate the activity of the membrane-bound form of an GPCRX protein or polypeptide or biologically-active portion thereof. The test compounds of the invention can be obtained using any of the numerous approaches in combinatorial library methods known in the art, including: biological libraries; spatially addressable parallel solid phase or solution phase libraries; synthetic library methods requiring deconvolution; the “one-bead one-compound” library method; and synthetic library methods using affinity chromatography selection. The biological library approach is limited to peptide libraries, while the other four approaches are applicable to peptide, non-peptide oligomer or small molecule libraries of compounds. See, e.g., Lam, 1997. Anticancer Drug Design 12: 145.

[0206] A “small molecule” as used herein, is meant to refer to a composition that has a molecular weight of less than about 5 kD and most preferably less than about 4 kD. Small molecules can be, e.g., nucleic acids, peptides, polypeptides, peptidomimetics, carbohydrates, lipids or other organic or inorganic molecules. Libraries of chemical and/or biological mixtures, such as fungal, bacterial, or algal extracts, are known in the art and can be screened with any of the assays of the invention.

[0207] Examples of methods for the synthesis of molecular libraries can be found in the art, for example in: DeWitt, et al., 1993. Proc. Natl. Acad. Sci. U.S.A. 90: 6909; Erb, et al., 1994. Proc. Natl. Acad. Sci. U.S.A. 91: 11422; Zuckermann, et al., 1994. J. Med. Chem. 37: 2678; Cho, et al., 1993. Science 261: 1303; Carrell, et al., 1994. Angew. Chem. Int. Ed. Engl. 33: 2059; Carell, et al., 1994. Angew. Chem. Int. Ed. Engl. 33: 2061; and Gallop, et al., 1994. J. Med. Chem. 37:1233.

[0208] Libraries of compounds may be presented in solution (e.g., Houghten, 1992. Biotechniques 13: 412-421), or on beads (Lam, 1991. Nature 354: 82-84), on chips (Fodor, 1993. Nature 364: 555-556), bacteria (Ladner, U.S. Pat. No. 5,223,409), spores (Ladner, U.S. Pat. No. 5,233,409), plasmids (Cull, et al., 1992. Proc. Natl. Acad. Sci. USA 89: 1865-1869) or on phage (Scott and Smith, 1990. Science 249: 386-390; Devlin, 1990. Science 249: 404406; Cwirla, et al., 1990. Proc. Natl. Acad. Sci. U.S.A. 87: 6378-6382; Felici, 1991. J. Mol. Biol. 222: 301-310; Ladner, U.S. Pat. No. 5,233,409.).

[0209] In one embodiment, an assay is a cell-based assay in which a cell which expresses a membrane-bound form of GPCRX protein, or a biologically-active portion thereof, on the cell surface is contacted with a test compound and the ability of the test compound to bind to an GPCRX protein determined. The cell, for example, can of mammalian origin or a yeast cell. Determining the ability of the test compound to bind to the GPCRX protein can be accomplished, for example, by coupling the test compound with a radioisotope or enzymatic label such that binding of the test compound to the GPCRX protein or biologically-active portion thereof can be determined by detecting the labeled compound in a complex. For example, test compounds can be labeled with 125I, 35S, 14C, or 3H, either directly or indirectly, and the radioisotope detected by direct counting of radioemission or by scintillation counting. Alternatively, test compounds can be enzymatically-labeled with, for example, horseradish peroxidase, alkaline phosphatase, or luciferase, and the enzymatic label detected by determination of conversion of an appropriate substrate to product. In one embodiment, the assay comprises contacting a cell which expresses a membrane-bound form of GPCRX protein, or a biologically-active portion thereof, on the cell surface with a known compound which binds GPCRX to form an assay mixture, contacting the assay mixture with a test compound, and determining the ability of the test compound to interact with an GPCRX protein, wherein determining the ability of the test compound to interact with an GPCRX protein comprises determining the ability of the test compound to preferentially bind to GPCRX protein or a biologically-active portion thereof as compared to the known compound.

[0210] In another embodiment, an assay is a cell-based assay comprising contacting a cell expressing a membrane-bound form of GPCRX protein, or a biologically-active portion thereof, on the cell surface with a test compound and determining the ability of the test compound to modulate (e.g., stimulate or inhibit) the activity of the GPCRX protein or biologically-active portion thereof. Determining the ability of the test compound to modulate the activity of GPCRX or a biologically-active portion thereof can be accomplished, for example, by determining the ability of the GPCRX protein to bind to or interact with an GPCRX target molecule. As used herein, a “target molecule” is a molecule with which an GPCRX protein binds or interacts in nature, for example, a molecule on the surface of a cell which expresses an GPCRX interacting protein, a molecule on the surface of a second cell, a molecule in the extracellular milieu, a molecule associated with the internal surface of a cell membrane or a cytoplasmic molecule. An GPCRX target molecule can be a non-GPCRX molecule or an GPCRX protein or polypeptide of the invention. In one embodiment, an GPCRX target molecule is a component of a signal transduction pathway that facilitates transduction of an extracellular signal (e.g. a signal generated by binding of a compound to a membrane-bound GPCRX molecule) through the cell membrane and into the cell. The target, for example, can be a second intercellular protein that has catalytic activity or a protein that facilitates the association of downstream signaling molecules with GPCRX.

[0211] Determining the ability of the GPCRX protein to bind to or interact with an GPCRX target molecule can be accomplished by one of the methods described above for determining direct binding. In one embodiment, determining the ability of the GPCRX protein to bind to or interact with an GPCRX target molecule can be accomplished by determining the activity of the target molecule. For example, the activity of the target molecule can be determined by detecting induction of a cellular second messenger of the target (i.e. intracellular Ca2+ diacylglycerol, IP3, etc.), detecting catalytic/enzymatic activity of the target an appropriate substrate, detecting the induction of a reporter gene (comprising an GPCRX-responsive regulatory element operatively linked to a nucleic acid encoding a detectable marker, e.g., luciferase), or detecting a cellular response, for example, cell survival, cellular differentiation, or cell proliferation.

[0212] In yet another embodiment, an assay of the invention is a cell-free assay comprising contacting an GPCRX protein or biologically-active portion thereof with a test compound and determining the ability of the test compound to bind to the GPCRX protein or biologically-active portion thereof. Binding of the test compound to the GPCRX protein can be determined either directly or indirectly as described above. In one such embodiment, the assay comprises contacting the GPCRX protein or biologically-active portion thereof with a known compound which binds GPCRX to form an assay mixture, contacting the assay mixture with a test compound, and determining the ability of the test compound to interact with an GPCRX protein, wherein determining the ability of the test compound to interact with an GPCRX protein comprises determining the ability of the test compound to preferentially bind to GPCRX or biologically-active portion thereof as compared to the known compound.

[0213] In still another embodiment, an assay is a cell-free assay comprising contacting GPCRX protein or biologically-active portion thereof with a test compound and determining the ability of the test compound to modulate (e.g. stimulate or inhibit) the activity of the GPCRX protein or biologically-active portion thereof. Determining the ability of the test compound to modulate the activity of GPCRX can be accomplished, for example, by determining the ability of the GPCRX protein to bind to an GPCRX target molecule by one of the methods described above for determining direct binding. In an alternative embodiment, determining the ability of the test compound to modulate the activity of GPCRX protein can be accomplished by determining the ability of the GPCRX protein further modulate an GPCRX target molecule. For example, the catalytic/enzymatic activity of the target molecule on an appropriate substrate can be determined as described, supra.

[0214] In yet another embodiment, the cell-free assay comprises contacting the GPCRX protein or biologically-active portion thereof with a known compound which binds GPCRX protein to form an assay mixture, contacting the assay mixture with a test compound, and determining the ability of the test compound to interact with an GPCRX protein, wherein determining the ability of the test compound to interact with an GPCRX protein comprises determining the ability of the GPCRX protein to preferentially bind to or modulate the activity of an GPCRX target molecule.

[0215] The cell-free assays of the invention are amenable to use of both the soluble form or the membrane-bound form of GPCRX protein. In the case of cell-free assays comprising the membrane-bound form of GPCRX protein, it may be desirable to utilize a solubilizing agent such that the membrane-bound form of GPCRX protein is maintained in solution. Examples of such solubilizing agents include non-ionic detergents such as n-octylglucoside, n-dodecylglucoside, n-dodecylmaltoside, octanoyl-N-methylglucamide, decanoyl-N-methylglucamide, Triton® X-100, Triton® X-114, Thesit®, Isotridecypoly(ethylene glycol ether)n, N-dodecyl-N,N-dimethyl-3-ammonio-1-propane sulfonate, 3-(3-cholamidopropyl) dimethylamminiol-1-propane sulfonate (CHAPS), or 3-(3-cholamidopropyl)dimethylamminiol-2-hydroxy-1-propane sulfonate (CHAPSO).

[0216] In more than one embodiment of the above assay methods of the invention, it may be desirable to immobilize either GPCRX protein or its target molecule to facilitate separation of complexed from uncomplexed forms of one or both of the proteins, as well as to accommodate automation of the assay. Binding of a test compound to GPCRX protein, or interaction of GPCRX protein with a target molecule in the presence and absence of a candidate compound, can be accomplished in any vessel suitable for containing the reactants. Examples of such vessels include microtiter plates, test tubes, and micro-centrifuge tubes. In one embodiment, a fusion protein can be provided that adds a domain that allows one or both of the proteins to be bound to a matrix. For example, GST-GPCRX fusion proteins or GST-target fusion proteins can be adsorbed onto glutathione sepharose beads (Sigma Chemical, St. Louis, Mo.) or glutathione derivatized microtiter plates, that are then combined with the test compound or the test compound and either the non-adsorbed target protein or GPCRX protein, and the mixture is incubated under conditions conducive to complex formation (e.g., at physiological conditions for salt and pH). Following incubation, the beads or microtiter plate wells are washed to remove any unbound components, the matrix immobilized in the case of beads, complex determined either directly or indirectly, for example, as described, supra. Alternatively, the complexes can be dissociated from the matrix, and the level of GPCRX protein binding or activity determined using standard techniques.

[0217] Other techniques for immobilizing proteins on matrices can also be used in the screening assays of the invention. For example, either the GPCRX protein or its target molecule can be immobilized utilizing conjugation of biotin and streptavidin. Biotinylated GPCRX protein or target molecules can be prepared from biotin-NHS (N-hydroxy-succinimide) using techniques well-known within the art (e.g., biotinylation kit, Pierce Chemicals, Rockford, Ill.), and immobilized in the wells of streptavidin-coated 96 well plates (Pierce Chemical). Alternatively, antibodies reactive with GPCRX protein or target molecules, but which do not interfere with binding of the GPCRX protein to its target molecule, can be derivatized to the wells of the plate, and unbound target or GPCRX protein trapped in the wells by antibody conjugation. Methods for detecting such complexes, in addition to those described above for the GST-immobilized complexes, include immunodetection of complexes using antibodies reactive with the GPCRX protein or target molecule, as well as enzyme-linked assays that rely on detecting an enzymatic activity associated with the GPCRX protein or target molecule.

[0218] In another embodiment, modulators of GPCRX protein expression are identified in a method wherein a cell is contacted with a candidate compound and the expression of GPCRX mRNA or protein in the cell is determined. The level of expression of GPCRX mRNA or protein in the presence of the candidate compound is compared to the level of expression of GPCRX mRNA or protein in the absence of the candidate compound. The candidate compound can then be identified as a modulator of GPCRX mRNA or protein expression based upon this comparison. For example, when expression of GPCRX mRNA or protein is greater (i.e., statistically significantly greater) in the presence of the candidate compound than in its absence, the candidate compound is identified as a stimulator of GPCRX mRNA or protein expression. Alternatively, when expression of GPCRX mRNA or protein is less (statistically significantly less) in the presence of the candidate compound than in its absence, the candidate compound is identified as an inhibitor of GPCRX mRNA or protein expression. The level of GPCRX mRNA or protein expression in the cells can be determined by methods described herein for detecting GPCRX mRNA or protein.

[0219] In yet another aspect of the invention, the GPCRX proteins can be used as “bait proteins” in a two-hybrid assay or three hybrid assay (see, e.g., U.S. Pat. No. 5,283,317; Zervos, et al., 1993. Cell 72: 223-232; Madura, et al., 1993. J. Biol. Chem. 268: 12046-12054; Bartel, et al., 1993. Biotechniques 14: 920-924; Iwabuchi, et al., 1993. Oncogene 8: 1693-1696; and Brent WO 94/10300), to identify other proteins that bind to or interact with GPCRX (“GPCRX-binding proteins” or “GPCRX-bp”) and modulate GPCRX activity. Such GPCRX-binding proteins are also likely to be involved in the propagation of signals by the GPCRX proteins as, for example, upstream or downstream elements of the GPCRX pathway.

[0220] The two-hybrid system is based on the modular nature of most transcription factors, which consist of separable DNA-binding and activation domains. Briefly, the assay utilizes two different DNA constructs. In one construct, the gene that codes for GPCRX is fused to a gene encoding the DNA binding domain of a known transcription factor (e.g. GALA). In the other construct, a DNA sequence, from a library of DNA sequences, that encodes an unidentified protein (“prey” or “sample”) is fused to a gene that codes for the activation domain of the known transcription factor. If the “bait” and the “prey” proteins are able to interact, in vivo, forming an GPCRX-dependent complex, the DNA-binding and activation domains of the transcription factor are brought into close proximity. This proximity allows transcription of a reporter gene (e.g., LacZ) that is operably linked to a transcriptional regulatory site responsive to the transcription factor. Expression of the reporter gene can be detected and cell colonies containing the functional transcription factor can be isolated and used to obtain the cloned gene that encodes the protein which interacts with GPCRX.

[0221] The invention further pertains to novel agents identified by the aforementioned screening assays and uses thereof for treatments as described herein.

[0222] Detection Assays

[0223] Portions or fragments of the cDNA sequences identified herein (and the corresponding complete gene sequences) can be used in numerous ways as polynucleotide reagents. By way of example, and not of limitation, these sequences can be used to: (i) map their respective genes on a chromosome; and, thus, locate gene regions associated with genetic disease; (ii) identify an individual from a minute biological sample (tissue typing); and (iii) aid in forensic identification of a biological sample. Some of these applications are described in the subsections, below.

[0224] Chromosome Mapping

[0225] Once the sequence (or a portion of the sequence) of a gene has been isolated, this sequence can be used to map the location of the gene on a chromosome. This process is called chromosome mapping. Accordingly, portions or fragments of the GPCRX sequences, SEQ ID NOS:1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, 75, 77, 79, 81, 83, 85, 87, 89, 91, 93, 95, 97, 99, 101, 103, 105, 107, 109, 111, 113, 115, 117, 119, 121, 123, 125, 127, 129, 131, 133, 135, 137, 139, 141, 143, 145, 147, 149 and 151, or fragments or derivatives thereof, can be used to map the location of the GPCRX genes, respectively, on a chromosome. The mapping of the GPCRX sequences to chromosomes is an important first step in correlating these sequences with genes associated with disease.

[0226] Briefly, GPCRX genes can be mapped to chromosomes by preparing PCR primers (preferably 15-25 bp in length) from the GPCRX sequences. Computer analysis of the GPCRX, sequences can be used to rapidly select primers that do not span more than one exon in the genomic DNA, thus complicating the amplification process. These primers can then be used for PCR screening of somatic cell hybrids containing individual human chromosomes. Only those hybrids containing the human gene corresponding to the GPCRX sequences will yield an amplified fragment.

[0227] Somatic cell hybrids are prepared by fusing somatic cells from different mammals (e.g., human and mouse cells). As hybrids of human and mouse cells grow and divide, they gradually lose human chromosomes in random order, but retain the mouse chromosomes. By using media in which mouse cells cannot grow, because they lack a particular enzyme, but in which human cells can, the one human chromosome that contains the gene encoding the needed enzyme will be retained. By using various media, panels of hybrid cell lines can be established. Each cell line in a panel contains either a single human chromosome or a small number of human chromosomes, and a full set of mouse chromosomes, allowing easy mapping of individual genes to specific human chromosomes. See, e.g., D'Eustachio, et al., 1983. Science 220: 919-924. Somatic cell hybrids containing only fragments of human chromosomes can also be produced by using human chromosomes with translocations and deletions.

[0228] PCR mapping of somatic cell hybrids is a rapid procedure for assigning a particular sequence to a particular chromosome. Three or more sequences can be assigned per day using a single thermal cycler. Using the GPCRX sequences to design oligonucleotide primers, sub-localization can be achieved with panels of fragments from specific chromosomes.

[0229] Fluorescence in situ hybridization (FISH) of a DNA sequence to a metaphase chromosomal spread can further be used to provide a precise chromosomal location in one step. Chromosome spreads can be made using cells whose division has been blocked in metaphase by a chemical like colcemid that disrupts the mitotic spindle. The chromosomes can be treated briefly with trypsin, and then stained with Giemsa. A pattern of light and dark bands develops on each chromosome, so that the chromosomes can be identified individually. The FISH technique can be used with a DNA sequence as short as 500 or 600 bases. However, clones larger than 1,000 bases have a higher likelihood of binding to a unique chromosomal location with sufficient signal intensity for simple detection. Preferably 1,000 bases, and more preferably 2,000 bases, will suffice to get good results at a reasonable amount of time. For a review of this technique, see, Verma, et al., HUMAN CHROMOSOMES: A MANUAL OF BASIC TECHNIQUES (Pergamon Press, New York 1988).

[0230] Reagents for chromosome mapping can be used individually to mark a single chromosome or a single site on that chromosome, or panels of reagents can be used for marking multiple sites and/or multiple chromosomes. Reagents corresponding to noncoding regions of the genes actually are preferred for mapping purposes. Coding sequences are more likely to be conserved within gene families, thus increasing the chance of cross hybridizations during chromosomal mapping.

[0231] Once a sequence has been mapped to a precise chromosomal location, the physical position of the sequence on the chromosome can be correlated with genetic map data. Such data are found, e.g., in McKusick, MENDELIAN INHERITANCE IN MAN, available on-line through Johns Hopkins University Welch Medical Library). The relationship between genes and disease, mapped to the same chromosomal region, can then be identified through linkage analysis (co-inheritance of physically adjacent genes), described in, e.g., Egeland, et al., 1987. Nature, 325: 783-787.

[0232] Moreover, differences in the DNA sequences between individuals affected and unaffected with a disease associated with the GPCRX gene, can be determined. If a mutation is observed in some or all of the affected individuals but not in any unaffected individuals, then the mutation is likely to be the causative agent of the particular disease. Comparison of affected and unaffected individuals generally involves first looking for structural alterations in the chromosomes, such as deletions or translocations that are visible from chromosome spreads or detectable using PCR based on that DNA sequence. Ultimately, complete sequencing of genes from several individuals can be performed to confirm the presence of a mutation and to distinguish mutations from polymorphisms.

[0233] Tissue Typing

[0234] The GPCRX sequences of the invention can also be used to identify individuals from minute biological samples. In this technique, an individual's genomic DNA is digested with one or more restriction enzymes, and probed on a Southern blot to yield unique bands for identification. The sequences of the invention are useful as additional DNA markers for RFLP (“restriction fragment length polymorphisms,” described in U.S. Pat. No. 5,272,057).

[0235] Furthermore, the sequences of the invention can be used to provide an alternative technique that determines the actual base-by-base DNA sequence of selected portions of an individual's genome. Thus, the GPCRX sequences described herein can be used to prepare two PCR primers from the 5′- and 3′-termini of the sequences. These primers can then be used to amplify an individual's DNA and subsequently sequence it.

[0236] Panels of corresponding DNA sequences from individuals, prepared in this manner, can provide unique individual identifications, as each individual will have a unique set of such DNA sequences due to allelic differences. The sequences of the invention can be used to obtain such identification sequences from individuals and from tissue. The GPCRX sequences of the invention uniquely represent portions of the human genome. Allelic variation occurs to some degree in the coding regions of these sequences, and to a greater degree in the noncoding regions. It is estimated that allelic variation between individual humans occurs with a frequency of about once per each 500 bases. Much of the allelic variation is due to single nucleotide polymorphisms (SNPs), which include restriction fragment length polymorphisms (RFLPs).

[0237] Each of the sequences described herein can, to some degree, be used as a standard against which DNA from an individual can be compared for identification purposes. Because greater numbers of polymorphisms occur in the noncoding regions, fewer sequences are necessary to differentiate individuals. The noncoding sequences can comfortably provide positive individual identification with a panel of perhaps 10 to 1,000 primers that each yield a noncoding amplified sequence of 100 bases. If predicted coding sequences, such as those in SEQ ID NOS:1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, 75, 77, 79, 81, 83, 85, 87, 89, 91, 93, 95, 97, 99, 101, 103, 105, 107, 109, 111, 113, 115, 117, 119, 121, 123, 125, 127, 129, 131, 133, 135, 137, 139, 141, 143, 145, 147, 149 and 151 are used, a more appropriate number of primers for positive individual identification would be 500-2,000.

[0238] Predictive Medicine

[0239] The invention also pertains to the field of predictive medicine in which diagnostic assays, prognostic assays, pharmacogenomics, and monitoring clinical trials are used for prognostic (predictive) purposes to thereby treat an individual prophylactically. Accordingly, one aspect of the invention relates to diagnostic assays for determining GPCRX protein and/or nucleic acid expression as well as GPCRX activity, in the context of a biological sample (e.g., blood, serum, cells, tissue) to thereby determine whether an individual is afflicted with a disease or disorder, or is at risk of developing a disorder, associated with aberrant GPCRX expression or activity. The disorders include metabolic disorders, diabetes, obesity, infectious disease, anorexia, cancer-associated cachexia, cancer, neurodegenerative disorders, Alzheimer's Disease, Parkinson's Disorder, immune disorders, and hematopoietic disorders, and the various dyslipidemias, metabolic disturbances associated with obesity, the metabolic syndrome X and wasting disorders associated with chronic diseases and various cancers. The invention also provides for prognostic (or predictive) assays for determining whether an individual is at risk of developing a disorder associated with GPCRX protein, nucleic acid expression or activity. For example, mutations in an GPCRX gene can be assayed in a biological sample. Such assays can be used for prognostic or predictive purpose to thereby prophylactically treat an individual prior to the onset of a disorder characterized by or associated with GPCRX protein, nucleic acid expression, or biological activity.

[0240] Another aspect of the invention provides methods for determining GPCRX protein, nucleic acid expression or activity in an individual to thereby select appropriate therapeutic or prophylactic agents for that individual (referred to herein as “pharmacogenomics”). Pharmacogenomics allows for the selection of agents (e.g., drugs) for therapeutic or prophylactic treatment of an individual based on the genotype of the individual (e.g. the genotype of the individual examined to determine the ability of the individual to respond to a particular agent.)

[0241] Yet another aspect of the invention pertains to monitoring the influence of agents (e.g., drugs, compounds) on the expression or activity of GPCRX in clinical trials.

[0242] These and other agents are described in further detail in the following sections.

[0243] Diagnostic Assays

[0244] An exemplary method for detecting the presence or absence of GPCRX in a biological sample involves obtaining a biological sample from a test subject and contacting the biological sample with a compound or an agent capable of detecting GPCRX protein or nucleic acid (e.g., mRNA, genomic DNA) that encodes GPCRX protein such that the presence of GPCRX is detected in the biological sample. An agent for detecting GPCRX mRNA or genomic DNA is a labeled nucleic acid probe capable of hybridizing to GPCRX mRNA or genomic DNA. The nucleic acid probe can be, for example, a full-length GPCRX nucleic acid, such as the nucleic acid of SEQ ID NOS:1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, 75, 77, 79, 81, 83, 85, 87, 89, 91, 93, 95, 97, 99, 101, 103, 105, 107, 109, 111, 113, 115, 117, 119, 121, 123, 125, 127, 129, 131, 133, 135, 137, 139, 141, 143, 145, 147, 149 and 151, or a portion thereof, such as an oligonucleotide of at least 15, 30, 50, 100, 250 or 500 nucleotides in length and sufficient to specifically hybridize under stringent conditions to GPCRX mRNA or genomic DNA. Other suitable probes for use in the diagnostic assays of the invention are described herein.

[0245] An agent for detecting GPCRX protein is an antibody capable of binding to GPCRX protein, preferably an antibody with a detectable label. Antibodies can be polyclonal, or more preferably, monoclonal. An intact antibody, or a fragment thereof (e.g., Fab or F(ab′)2) can be used. The term “labeled”, with regard to the probe or antibody, is intended to encompass direct labeling of the probe or antibody by coupling (i.e., physically linking) a detectable substance to the probe or antibody, as well as indirect labeling of the probe or antibody by reactivity with another reagent that is directly labeled. Examples of indirect labeling include detection of a primary antibody using a fluorescently-labeled secondary antibody and end-labeling of a DNA probe with biotin such that it can be detected with fluorescently-labeled streptavidin. The term “biological sample” is intended to include tissues, cells and biological fluids isolated from a subject, as well as tissues, cells and fluids present within a subject. That is, the detection method of the invention can be used to detect GPCRX mRNA, protein, or genomic DNA in a biological sample in vitro as well as in vivo. For example, in vitro techniques for detection of GPCRX mRNA include Northern hybridizations and in situ hybridizations. In vitro techniques for detection of GPCRX protein include enzyme linked immunosorbent assays (ELISAs), Western blots, immunoprecipitations, and immunofluorescence. In vitro techniques for detection of GPCRX genomic DNA include Southern hybridizations. Furthermore, in vivo techniques for detection of GPCRX protein include introducing into a subject a labeled anti-GPCRX antibody. For example, the antibody can be labeled with a radioactive marker whose presence and location in a subject can be detected by standard imaging techniques.

[0246] In one embodiment, the biological sample contains protein molecules from the test subject. Alternatively, the biological sample can contain mRNA molecules from the test subject or genomic DNA molecules from the test subject. A preferred biological sample is a peripheral blood leukocyte sample isolated by conventional means from a subject.

[0247] In another embodiment, the methods further involve obtaining a control biological sample from a control subject, contacting the control sample with a compound or agent capable of detecting GPCRX protein, mRNA, or genomic DNA, such that the presence of GPCRX protein, mRNA or genomic DNA is detected in the biological sample, and comparing the presence of GPCRX protein, mRNA or genomic DNA in the control sample with the presence of GPCRX protein, mRNA or genomic DNA in the test sample.

[0248] The invention also encompasses kits for detecting the presence of GPCRX in a biological sample. For example, the kit can comprise: a labeled compound or agent capable of detecting GPCRX protein or mRNA in a biological sample; means for determining the amount of GPCRX in the sample; and means for comparing the amount of GPCRX in the sample with a standard. The compound or agent can be packaged in a suitable container. The kit can further comprise instructions for using the kit to detect GPCRX protein or nucleic acid.

[0249] Prognostic Assays

[0250] The diagnostic methods described herein can furthermore be utilized to identify subjects having or at risk of developing a disease or disorder associated with aberrant GPCRX expression or activity. For example, the assays described herein, such as the preceding diagnostic assays or the following assays, can be utilized to identify a subject having or at risk of developing a disorder associated with GPCRX protein, nucleic acid expression or activity. Alternatively, the prognostic assays can be utilized to identify a subject having or at risk for developing a disease or disorder. Thus, the invention provides a method for identifying a disease or disorder associated with aberrant GPCRX expression or activity in which a test sample is obtained from a subject and GPCRX protein or nucleic acid (e.g., mRNA, genomic DNA) is detected, wherein the presence of GPCRX protein or nucleic acid is diagnostic for a subject having or at risk of developing a disease or disorder associated with aberrant GPCRX expression or activity. As used herein, a “test sample” refers to a biological sample obtained from a subject of interest. For example, a test sample can be a biological fluid (e.g., serum), cell sample, or tissue.

[0251] Furthermore, the prognostic assays described herein can be used to determine whether a subject can be administered an agent (e.g., an agonist, antagonist, peptidomimetic, protein, peptide, nucleic acid, small molecule, or other drug candidate) to treat a disease or disorder associated with aberrant GPCRX expression or activity. For example, such methods can be used to determine whether a subject can be effectively treated with an agent for a disorder. Thus, the invention provides methods for determining whether a subject can be effectively treated with an agent for a disorder associated with aberrant GPCRX expression or activity in which a test sample is obtained and GPCRX protein or nucleic acid is detected (e.g. wherein the presence of GPCRX protein or nucleic acid is diagnostic for a subject that can be administered the agent to treat a disorder associated with aberrant GPCRX expression or activity).

[0252] The methods of the invention can also be used to detect genetic lesions in an GPCRX gene, thereby determining if a subject with the lesioned gene is at risk for a disorder characterized by aberrant cell proliferation and/or differentiation. In various embodiments, the methods include detecting, in a sample of cells from the subject, the presence or absence of a genetic lesion characterized by at least one of an alteration affecting the integrity of a gene encoding an GPCRX-protein, or the misexpression of the GPCRX gene. For example, such genetic lesions can be detected by ascertaining the existence of at least one of: (i) a deletion of one or more nucleotides from an GPCRX gene; (ii) an addition of one or more nucleotides to an GPCRX gene; (iii) a substitution of one or more nucleotides of an GPCRX gene, (iv) a chromosomal rearrangement of an GPCRX gene; (v) an alteration in the level of a messenger RNA transcript of an GPCRX gene, (vi) aberrant modification of an GPCRX gene, such as of the methylation pattern of the genomic DNA, (vii) the presence of a non-wild-type splicing pattern of a messenger RNA transcript of an GPCRX gene, (viii) a non-wild-type level of an GPCRX protein, (ix) allelic loss of an GPCRX gene, and (x) inappropriate post-translational modification of an GPCRX protein. As described herein, there are a large number of assay techniques known in the art which can be used for detecting lesions in an GPCRX gene. A preferred biological sample is a peripheral blood leukocyte sample isolated by conventional means from a subject. However, any biological sample containing nucleated cells may be used, including, for example, buccal mucosal cells.

[0253] In certain embodiments, detection of the lesion involves the use of a probe/primer in a polymerase chain reaction (PCR) (see, e.g., U.S. Pat. Nos. 4,683,195 and 4,683,202), such as anchor PCR or RACE PCR, or, alternatively, in a ligation chain reaction (LCR) (see, e.g., Landegran, et al., 1988. Science 241: 1077-1080; and Nakazawa, et al., 1994. Proc. Natl. Acad. Sci. USA 91: 360-364), the latter of which can be particularly useful for detecting point mutations in the GPCRX-gene (see, Abravaya, et al., 1995. Nucl. Acids Res. 23: 675-682). This method can include the steps of collecting a sample of cells from a patient, isolating nucleic acid (e.g., genomic, mRNA or both) from the cells of the sample, contacting the nucleic acid sample with one or more primers that specifically hybridize to an GPCRX gene under conditions such that hybridization and amplification of the GPCRX gene (if present) occurs, and detecting the presence or absence of an amplification product, or detecting the size of the amplification product and comparing the length to a control sample. It is anticipated that PCR and/or LCR may be desirable to use as a preliminary amplification step in conjunction with any of the techniques used for detecting mutations described herein.

[0254] Alternative amplification methods include: self sustained sequence replication (see, Guatelli, et al., 1990. Proc. Natl. Acad. Sci. USA 87: 1874-1878), transcriptional amplification system (see, Kwoh, et al., 1989. Proc. Natl. Acad. Sci. USA 86: 1173-1177); Qβ Replicase (see, Lizardi, et al, 1988. BioTechnology 6: 1197), or any other nucleic acid amplification method, followed by the detection of the amplified molecules using techniques well known to those of skill in the art. These detection schemes are especially useful for the detection of nucleic acid molecules if such molecules are present in very low numbers.

[0255] In an alternative embodiment, mutations in an GPCRX gene from a sample cell can be identified by alterations in restriction enzyme cleavage patterns. For example, sample and control DNA is isolated, amplified (optionally), digested with one or more restriction endonucleases, and fragment length sizes are determined by gel electrophoresis and compared. Differences in fragment length sizes between sample and control DNA indicates mutations in the sample DNA. Moreover, the use of sequence specific ribozymes (see, e.g., U.S. Pat. No. 5,493,531) can be used to score for the presence of specific mutations by development or loss of a ribozyme cleavage site.

[0256] In other embodiments, genetic mutations in GPCRX can be identified by hybridizing a sample and control nucleic acids, e.g., DNA or RNA, to high-density arrays containing hundreds or thousands of oligonucleotides probes. See, e.g., Cronin, et al., 1996. Human Mutation 7: 244-255; Kozal, et al., 1996. Nat. Med. 2: 753-759. For example, genetic mutations in GPCRX can be identified in two dimensional arrays containing light-generated DNA probes as described in Cronin, et al., supra. Briefly, a first hybridization array of probes can be used to scan through long stretches of DNA in a sample and control to identify base changes between the sequences by making linear arrays of sequential overlapping probes. This step allows the identification of point mutations. This is followed by a second hybridization array that allows the characterization of specific mutations by using smaller, specialized probe arrays complementary to all variants or mutations detected. Each mutation array is composed of parallel probe sets, one complementary to the wild-type gene and the other complementary to the mutant gene.

[0257] In yet another embodiment, any of a variety of sequencing reactions known in the art can be used to directly sequence the GPCRX gene and detect mutations by comparing the sequence of the sample GPCRX with the corresponding wild-type (control) sequence. Examples of sequencing reactions include those based on techniques developed by Maxim and Gilbert, 1977. Proc. Natl. Acad. Sci. USA 74: 560 or Sanger, 1977. Proc. Natl. Acad. Sci. USA 74: 5463. It is also contemplated that any of a variety of automated sequencing procedures can be utilized when performing the diagnostic assays (see, e.g. Naeve, et al., 1995. Biotechniques 19: 448), including sequencing by mass spectrometry (see, e.g., PCT International Publication No. WO 94/16101; Cohen, et al., 1996. Adv. Chromatography 36: 127-162; and Griffin, et al., 1993. Appl. Biochem. Biotechnol. 38: 147-159).

[0258] Other methods for detecting mutations in the GPCRX gene include methods in which protection from cleavage agents is used to detect mismatched bases in RNA/RNA or RNA/DNA heteroduplexes. See, e.g., Myers, et al., 1985. Science 230: 1242. In general, the art technique of “mismatch cleavage” starts by providing heteroduplexes of formed by hybridizing (labeled) RNA or DNA containing the wild-type GPCRX sequence with potentially mutant RNA or DNA obtained from a tissue sample. The double-stranded duplexes are treated with an agent that cleaves single-stranded regions of the duplex such as which will exist due to basepair mismatches between the control and sample strands. For instance, RNA/DNA duplexes can be treated with RNase and DNA/DNA hybrids treated with S1 nuclease to enzymatically digesting the mismatched regions. In other embodiments, either DNA/DNA or RNA/DNA duplexes can be treated with hydroxylamine or osmium tetroxide and with piperidine in order to digest mismatched regions. After digestion of the mismatched regions, the resulting material is then separated by size on denaturing polyacrylamide gels to determine the site of mutation. See, e.g., Cotton, et al., 1988. Proc. Natl. Acad. Sci. USA 85: 4397; Saleeba, et al., 1992. Methods Enzymol. 217: 286-295. In an embodiment, the control DNA or RNA can be labeled for detection.

[0259] In still another embodiment, the mismatch cleavage reaction employs one or more proteins that recognize mismatched base pairs in double-stranded DNA (so called “DNA mismatch repair” enzymes) in defined systems for detecting and mapping point mutations in GPCRX cDNAs obtained from samples of cells. For example, the mutY enzyme of E. coli cleaves A at G/A mismatches and the thymidine DNA glycosylase from HeLa cells cleaves T at G/T mismatches. See, e.g., Hsu, et al., 1994. Carcinogenesis 15: 1657-1662. According to an exemplary embodiment, a probe based on an GPCRX sequence, e.g., a wild-type GPCRX sequence, is hybridized to a cDNA or other DNA product from a test cell(s). The duplex is treated with a DNA mismatch repair enzyme, and the cleavage products, if any, can be detected from electrophoresis protocols or the like. See, e.g., U.S. Pat. No. 5,459,039.

[0260] In other embodiments, alterations in electrophoretic mobility will be used to identify mutations in GPCRX genes. For example, single strand conformation polymorphism (SSCP) may be used to detect differences in electrophoretic mobility between mutant and wild type nucleic acids. See, e.g., Orita, et al., 1989. Proc. Natl. Acad. Sci. USA: 86: 2766; Cotton, 1993. Mutat. Res. 285: 125-144; Hayashi, 1992. Genet. Anal. Tech. Appl. 9: 73-79. Single-stranded DNA fragments of sample and control GPCRX nucleic acids will be denatured and allowed to renature. The secondary structure of single-stranded nucleic acids varies according to sequence, the resulting alteration in electrophoretic mobility enables the detection of even a single base change. The DNA fragments may be labeled or detected with labeled probes. The sensitivity of the assay may be enhanced by using RNA (rather than DNA), in which the secondary structure is more sensitive to a change in sequence. In one embodiment, the subject method utilizes heteroduplex analysis to separate double stranded heteroduplex molecules on the basis of changes in electrophoretic mobility. See, e.g., Keen, et al., 1991. Trends Genet. 7: 5.

[0261] In yet another embodiment, the movement of mutant or wild-type fragments in polyacrylamide gels containing a gradient of denaturant is assayed using denaturing gradient gel electrophoresis (DGGE). See, e.g., Myers, et al., 1985. Nature 313: 495. When DGGE is used as the method of analysis, DNA will be modified to insure that it does not completely denature, for example by adding a GC clamp of approximately 40 bp of high-melting GC-rich DNA by PCR. In a further embodiment, a temperature gradient is used in place of a denaturing gradient to identify differences in the mobility of control and sample DNA. See, e.g., Rosenbaum and Reissner, 1987. Biophys. Chem. 265: 12753.

[0262] Examples of other techniques for detecting point mutations include, but are not limited to, selective oligonucleotide hybridization, selective amplification, or selective primer extension. For example, oligonucleotide primers may be prepared in which the known mutation is placed centrally and then hybridized to target DNA under conditions that permit hybridization only if a perfect match is found. See, e.g., Saiki, et al., 1986. Nature 324: 163; Saiki, et al., 1989. Proc. Natl. Acad. Sci. USA 86: 6230. Such allele specific oligonucleotides are hybridized to PCR amplified target DNA or a number of different mutations when the oligonucleotides are attached to the hybridizing membrane and hybridized with labeled target DNA.

[0263] Alternatively, allele specific amplification technology that depends on selective PCR amplification may be used in conjunction with the instant invention. Oligonucleotides used as primers for specific amplification may carry the mutation of interest in the center of the molecule (so that amplification depends on differential hybridization; see, e.g., Gibbs, et al., 1989. Nucl. Acids Res. 17: 2437-2448) or at the extreme 3′-terminus of one primer where, under appropriate conditions, mismatch can prevent, or reduce polymerase extension (see, e.g., Prossner, 1993. Tibtech. 11: 238). In addition it may be desirable to introduce a novel restriction site in the region of the mutation to create cleavage-based detection. See, e.g., Gasparini, et al., 1992. Mol. Cell Probes 6: 1. It is anticipated that in certain embodiments amplification may also be performed using Taq ligase for amplification. See, e.g. Barany, 1991. Proc. Natl. Acad. Sci USA 88: 189. In such cases, ligation will occur only if there is a perfect match at the 3′-terminus of the 5′ sequence, making it possible to detect the presence of a known mutation at a specific site by looking for the presence or absence of amplification.

[0264] The methods described herein may be performed, for example, by utilizing pre-packaged diagnostic kits comprising at least one probe nucleic acid or antibody reagent described herein, which may be conveniently used, e.g., in clinical settings to diagnose patients exhibiting symptoms or family history of a disease or illness involving an GPCRX gene.

[0265] Furthermore, any cell type or tissue, preferably peripheral blood leukocytes, in which GPCRX is expressed may be utilized in the prognostic assays described herein. However, any biological sample containing nucleated cells may be used, including, for example, buccal mucosal cells.

[0266] Pharmacogenomics

[0267] Agents, or modulators that have a stimulatory or inhibitory effect on GPCRX activity (e.g., GPCRX gene expression), as identified by a screening assay described herein can be administered to individuals to treat (prophylactically or therapeutically) disorders (The disorders include metabolic disorders, diabetes, obesity, infectious disease, anorexia, cancer-associated cachexia, cancer, neurodegenerative disorders, Alzheimer's Disease, Parkinson's Disorder, immune disorders, and hematopoietic disorders, and the various dyslipidemias, metabolic disturbances associated with obesity, the metabolic syndrome X and wasting disorders associated with chronic diseases and various cancers.) In conjunction with such treatment, the pharmacogenomics (i.e., the study of the relationship between an individual's genotype and that individual's response to a foreign compound or drug) of the individual may be considered. Differences in metabolism of therapeutics can lead to severe toxicity or therapeutic failure by altering the relation between dose and blood concentration of the pharmacologically active drug. Thus, the pharmacogenomics of the individual permits the selection of effective agents (e.g., drugs) for prophylactic or therapeutic treatments based on a consideration of the individual's genotype. Such pharmacogenomics can further be used to determine appropriate dosages and therapeutic regimens. Accordingly, the activity of GPCRX protein, expression of GPCRX nucleic acid, or mutation content of GPCRX genes in an individual can be determined to thereby select appropriate agent(s) for therapeutic or prophylactic treatment of the individual.

[0268] Pharmacogenomics deals with clinically significant hereditary variations in the response to drugs due to altered drug disposition and abnormal action in affected persons. See e.g., Eichelbaum, 1996. Clin. Exp. Pharmacol. Physiol., 23: 983-985; Linder, 1997. Clin. Chem., 43: 254-266. In general, two types of pharmacogenetic conditions can be -differentiated. Genetic conditions transmitted as a single factor altering the way drugs act on the body (altered drug action) or genetic conditions transmitted as single factors altering the way the body acts on drugs (altered drug metabolism). These pharmacogenetic conditions can occur either as rare defects or as polymorphisms. For example, glucose-6-phosphate dehydrogenase (G6PD) deficiency is a common inherited enzymopathy in which the main clinical complication is hemolysis after ingestion of oxidant drugs (anti-malarials, sulfonamides, analgesics, nitrofurans) and consumption of fava beans.

[0269] As an illustrative embodiment, the activity of drug metabolizing enzymes is a major determinant of both the intensity and duration of drug action. The discovery of genetic polymorphisms of drug metabolizing enzymes (e.g., N-acetyltransferase 2 (NAT 2) and cytochrome P450 enzymes CYP2D6 and CYP2C 19) has provided an explanation as to why some patients do not obtain the expected drug effects or show exaggerated drug response and serious toxicity after taking the standard and safe dose of a drug. These polymorphisms are expressed in two phenotypes in the population, the extensive metabolizer (EM) and poor metabolizer (PM). The prevalence of PM is different among different populations. For example, the gene coding for CYP2D6 is highly polymorphic and several mutations have been identified in PM, which all lead to the absence of functional CYP2D6. Poor metabolizers of CYP2D6 and CYP2C 19 quite frequently experience exaggerated drug response and side effects when they receive standard doses. If a metabolite is the active therapeutic moiety, PM show no therapeutic response, as demonstrated for the analgesic effect of codeine mediated by its CYP2D6-formed metabolite morphine. At the other extreme are the so called ultra-rapid metabolizers who do not respond to standard doses. Recently, the molecular basis of ultra-rapid metabolism has been identified to be due to CYP2D6 gene amplification.

[0270] Thus, the activity of GPCRX protein, expression of GPCRX nucleic acid, or mutation content of GPCRX genes in an individual can be determined to thereby select appropriate agent(s) for therapeutic or prophylactic treatment of the individual. In addition, pharmacogenetic studies can be used to apply genotyping of polymorphic alleles encoding drug-metabolizing enzymes to the identification of an individual's drug responsiveness phenotype. This knowledge, when applied to dosing or drug selection, can avoid adverse reactions or therapeutic failure and thus enhance therapeutic or prophylactic efficiency when treating a subject with an GPCRX modulator, such as a modulator identified by one of the exemplary screening assays described herein.

[0271] Monitoring of Effects During Clinical Trials

[0272] Monitoring the influence of agents (e.g., drugs, compounds) on the expression or activity of GPCRX (e.g., the ability to modulate aberrant cell proliferation and/or differentiation) can be applied not only in basic drug screening, but also in clinical trials. For example, the effectiveness of an agent determined by a screening assay as described herein to increase GPCRX gene expression, protein levels, or upregulate GPCRX activity, can be monitored in clinical trails of subjects exhibiting decreased GPCRX gene expression, protein levels, or downregulated GPCRX activity. Alternatively, the effectiveness of an agent determined by a screening assay to decrease GPCRX gene expression, protein levels, or downregulate GPCRX activity, can be monitored in clinical trails of subjects exhibiting increased GPCRX gene expression, protein levels, or upregulated GPCRX activity. In such clinical trials, the expression or activity of GPCRX and, preferably, other genes that have been implicated in, for example, a cellular proliferation or immune disorder can be used as a “read out” or markers of the immune responsiveness of a particular cell.

[0273] By way of example, and not of limitation, genes, including GPCRX, that are modulated in cells by treatment with an agent (e.g., compound, drug or small molecule) that modulates GPCRX activity (e.g., identified in a screening assay as described herein) can be identified. Thus, to study the effect of agents on cellular proliferation disorders, for example, in a clinical trial, cells can be isolated and RNA prepared and analyzed for the levels of expression of GPCRX and other genes implicated in the disorder. The levels of gene expression (i.e., a gene expression pattern) can be quantified by Northern blot analysis or RT-PCR, as described herein, or alternatively by measuring the amount of protein produced, by one of the methods as described herein, or by measuring the levels of activity of GPCRX or other genes. In this manner, the gene expression pattern can serve as a marker, indicative of the physiological response of the cells to the agent. Accordingly, this response state may be determined before, and at various points during, treatment of the individual with the agent.

[0274] In one embodiment, the invention provides a method for monitoring the effectiveness of treatment of a subject with an agent (e.g., an agonist, antagonist, protein, peptide, peptidomimetic, nucleic acid, small molecule, or other drug candidate identified by the screening assays described herein) comprising the steps of (i) obtaining a pre-administration sample from a subject prior to administration of the agent; (ii) detecting the level of expression of an GPCRX protein, mRNA, or genomic DNA in the preadministration sample; (iii) obtaining one or more post-administration samples from the subject; (iv) detecting the level of expression or activity of the GPCRX protein, mRNA, or genomic DNA in the post-administration samples; (v) comparing the level of expression or activity of the GPCRX protein, mRNA, or genomic DNA in the pre-administration sample with the GPCRX protein, mRNA, or genomic DNA in the post administration sample or samples; and (vi) altering the administration of the agent to the subject accordingly. For example, increased administration of the agent may be desirable to increase the expression or activity of GPCRX to higher levels than detected, i.e., to increase the effectiveness of the agent. Alternatively, decreased administration of the agent may be desirable to decrease expression or activity of GPCRX to lower levels than detected, i.e., to decrease the effectiveness of the agent.

[0275] Methods of Treatment

[0276] The invention provides for both prophylactic and therapeutic methods of treating a subject at risk of (or susceptible to) a disorder or having a disorder associated with aberrant GPCRX expression or activity. The disorders include cardiomyopathy, atherosclerosis, hypertension, congenital heart defects, aortic stenosis, atrial septal defect (ASD), atrioventricular (A-V) canal defect, ductus arteriosus, pulmonary stenosis, subaortic stenosis, ventricular septal defect (VSD), valve diseases, tuberous sclerosis, scleroderma, obesity, transplantation, adrenoleukodystrophy, congenital adrenal hyperplasia, prostate cancer, neoplasm; adenocarcinoma, lymphoma, uterus cancer, fertility, hemophilia, hypercoagulation, idiopathic thrombocytopenic purpura, immunodeficiencies, graft versus host disease, AIDS, bronchial asthma, Crohn's disease; multiple sclerosis, treatment of Albright Hereditary Ostoeodystrophy, and other diseases, disorders and conditions of the like.

[0277] These methods of treatment will be discussed more fully, below.

[0278] Disease and Disorders

[0279] Diseases and disorders that are characterized by increased (relative to a subject not suffering from the disease or disorder) levels or biological activity may be treated with Therapeutics that antagonize (i.e., reduce or inhibit) activity. Therapeutics that antagonize activity may be administered in a therapeutic or prophylactic manner. Therapeutics that may be utilized include, but are not limited to: (i) an aforementioned peptide, or analogs, derivatives, fragments or homologs thereof; (ii) antibodies to an aforementioned peptide; (iii) nucleic acids encoding an aforementioned peptide; (iv) administration of antisense nucleic acid and nucleic acids that are “dysfunctional” (i.e., due to a heterologous insertion within the coding sequences of coding sequences to an aforementioned peptide) that are utilized to =“knockout” endoggenous function of an aforementioned peptide by homologous recombination (see, e.g., Capecchi, 1989. Science 244: 1288-1292); or (v) modulators (i.e., inhibitors, agonists and antagonists, including additional peptide mimetic of the invention or antibodies specific to a peptide of the invention) that alter the interaction between an aforementioned peptide and its binding partner.

[0280] Diseases and disorders that are characterized by decreased (relative to a subject not suffering from the disease or disorder) levels or biological activity may be treated with Therapeutics that increase (ie., are agonists to) activity. Therapeutics that upregulate activity may be administered in a therapeutic or prophylactic manner. Therapeutics that may be utilized include, but are not limited to, an aforementioned peptide, or analogs, derivatives, fragments or homologs thereof, or an agonist that increases bioavailability.

[0281] Increased or decreased levels can be readily detected by quantifying peptide and/or RNA, by obtaining a patient tissue sample (e.g., from biopsy tissue) and assaying it in vitro for RNA or peptide levels, structure and/or activity of the expressed peptides (or mRNAs of an aforementioned peptide). Methods that are well-known within the art include, but are not limited to, immunoassays (e.g., by Western blot analysis, immunoprecipitation followed by sodium dodecyl sulfate (SDS) polyacrylamide gel electrophoresis, immunocytochemistry, etc.) and/or hybridization assays to detect expression of mRNAs (e.g., Northern assays, dot blots, in situ hybridization, and the like).

[0282] Prophylactic Methods

[0283] In one aspect, the invention provides a method for preventing, in a subject, a disease or condition associated with an aberrant GPCRX expression or activity, by administering to the subject an agent that modulates GPCRX expression or at least one GPCRX activity. Subjects at risk for a disease that is caused or contributed to by aberrant GPCRX expression or activity can be identified by, for example, any or a combination of diagnostic or prognostic assays as described herein. Administration of a prophylactic agent can occur prior to the manifestation of symptoms characteristic of the GPCRX aberrancy, such that a disease or disorder is prevented or, alternatively, delayed in its progression. Depending upon the type of GPCRX aberrancy, for example, an GPCRX agonist or GPCRX antagonist agent can be used for treating the subject. The appropriate agent can be determined based on screening assays described herein. The prophylactic methods of the invention are further discussed in the following subsections.

[0284] Therapeutic Methods

[0285] Another aspect of the invention pertains to methods of modulating GPCRX expression or activity for therapeutic purposes. The modulatory method of the invention involves contacting a cell with an agent that modulates one or more of the activities of GPCRX protein activity associated with the cell. An agent that modulates GPCRX protein activity can be an agent as described herein, such as a nucleic acid or a protein, a naturally-occurring cognate ligand of an GPCRX protein, a peptide, an GPCRX peptidomimetic, or other small molecule. In one embodiment, the agent stimulates one or more GPCRX protein activity. Examples of such stimulatory agents include active GPCRX protein and a nucleic acid molecule encoding GPCRX that has been introduced into the cell. In another embodiment, the agent inhibits one or more GPCRX protein activity. Examples of such inhibitory agents include antisense GPCRX nucleic acid molecules and anti-GPCRX antibodies. These modulatory methods can be performed in vitro (e.g., by culturing the cell with the agent) or, alternatively, in vivo (e.g., by administering the agent to a subject). As such, the invention provides methods of treating an individual afflicted with a disease or disorder characterized by aberrant expression or activity of an GPCRX protein or nucleic acid molecule. In one embodiment, the method involves administering an agent (e.g., an agent identified by a screening assay described herein), or combination of agents that modulates (e.g., up-regulates or down-regulates) GPCRX expression or activity. In another embodiment, the method involves administering an GPCRX protein or nucleic acid molecule as therapy to compensate for reduced or aberrant GPCRX expression or activity.

[0286] Stimulation of GPCRX activity is desirable in situations in which GPCRX is abnormally downregulated and/or in which increased GPCRX activity is likely to have a beneficial effect. One example of such a situation is where a subject has a disorder characterized by aberrant cell proliferation and/or differentiation (e.g., cancer or immune associated disorders). Another example of such a situation is where the subject has a gestational disease (e.g., preclampsia).

[0287] Determination of the Biological Effect of the Therapeutic

[0288] In various embodiments of the invention, suitable in vitro or in vivo assays are performed to determine the effect of a specific Therapeutic and whether its administration is indicated for treatment of the affected tissue.

[0289] In various specific embodiments, in vitro assays may be performed with representative cells of the type(s) involved in the patient's disorder, to determine if a given Therapeutic exerts the desired effect upon the cell type(s). Compounds for use in therapy may be tested in suitable animal model systems including, but not limited to rats, mice, chicken, cows, monkeys, rabbits, and the like, prior to testing in human subjects. Similarly, for in vivo testing, any of the animal model system known in the art may be used prior to administration to human subjects.

[0290] Prophylactic and Therapeutic Uses of the Compositions of the Invention

[0291] The GPCRX nucleic acids and proteins of the invention are useful in potential prophylactic and therapeutic applications implicated in a variety of disorders including, but not limited to: metabolic disorders, diabetes, obesity, infectious disease, anorexia, cancer-associated cancer, neurodegenerative disorders, Alzheimer's Disease, Parkinson's Disorder, immune disorders, hematopoietic disorders, and the various dyslipidemias, metabolic disturbances associated with obesity, the metabolic syndrome X and wasting disorders associated with chronic diseases and various cancers.

[0292] As an example, a cDNA encoding the GPCRX protein of the invention may be useful in gene therapy, and the protein may be useful when administered to a subject in need thereof. By way of non-limiting example, the compositions of the invention will have efficacy for treatment of patients suffering from: metabolic disorders, diabetes, obesity, infectious disease, anorexia, cancer-associated cachexia, cancer, neurodegenerative disorders, Alzheimer's Disease, Parkinson's Disorder, immune disorders, hematopoietic disorders, and the various dyslipidemias.

[0293] Both the novel nucleic acid encoding the GPCRX protein, and the GPCRX protein of the invention, or fragments thereof, may also be useful in diagnostic applications, wherein the presence or amount of the nucleic acid or the protein are to be assessed. A further use could be as an anti-bacterial molecule (i.e., some peptides have been found to possess anti-bacterial properties). These materials are further useful in the generation of antibodies which immunospecifically-bind to the novel substances of the invention for use in therapeutic or diagnostic methods.

[0294] The invention will be further described in the following examples, which do not limit the scope of the invention described in the claims.

EXAMPLES

Example 1

Identification of GPCRX Nucleic Acids

[0295] TblastN using CuraGen Corporation's sequence file for polypeptides or homologs was run against the Genomic Daily Files made available by GenBank or from files downloaded from the individual sequencing centers. Exons were predicted by homology and the intron/exon boundaries were determined using standard genetic rules. Exons were further selected and refined by means of similarity determination using multiple BLAST (for example, tBlastN, BlastX, and BlastN) searches, and, in some instances, GeneScan and Grail. Expressed sequences from both public and proprietary databases were also added when available to further define and complete the gene sequence. The DNA sequence was then manually corrected for apparent inconsistencies thereby obtaining the sequences encoding the full-length protein.

[0296] The novel GPCRX target sequences identified in the present invention were subjected to the exon linking process to confirm the sequence. PCR primers were designed by starting at the most upstream sequence available, for the forward primer, and at the most downstream sequence available for the reverse primer. PCR primer sequences were used for obtaining different clones. In each case, the sequence was examined, walking inward from the respective termini toward the coding sequence, until a suitable sequence that is either unique or highly selective was encountered, or, in the case of the reverse primer, until the stop codon was reached. Such primers were designed based on in silico predictions for the full length cDNA, part (one or more exons) of the DNA or protein sequence of the target sequence, or by translated homology of the predicted exons to closely related human sequences from other species. These primers were then employed in PCR amplification based on the following pool of human cDNAs: adrenal gland, bone marrow, brain—amygdala, brain—cerebellum, brain—hippocampus, brain—substantia nigra, brain—thalamus, brain—whole, fetal brain, fetal kidney, fetal liver, fetal lung, heart, kidney, lymphoma—Raji, mammary gland, pancreas, pituitary gland, placenta, prostate, salivary gland, skeletal muscle, small intestine, spinal cord, spleen, stomach, testis, thyroid, trachea, uterus. Usually the resulting amplicons were gel purified, cloned and sequenced to high redundancy. The PCR product derived from exon linking was cloned into the pCR2.1 vector from Invitrogen. The resulting bacterial clone has an insert covering the entire open reading frame cloned into the pCR2.1 vector. The resulting sequences from all clones were assembled with themselves, with other fragments in CuraGen Corporation's database and with public ESTs. Fragments and ESTs were included as components for an assembly when the extent of their identity with another component of the assembly was at least 95% over 50 bp. In addition, sequence traces were evaluated manually and edited for corrections if appropriate. These procedures provide the sequence reported herein.

[0297] Physical clone: Exons were predicted by homology and the intron/exon boundaries were determined using standard genetic rules. Exons were further selected and refined by means of similarity determination using multiple BLAST (for example, tBlastN, BlastX, and BlastN) searches, and, in some instances, GeneScan and Grail. Expressed sequences from both public and proprietary databases were also added when available to further define and complete the gene sequence. The DNA sequence was then manually corrected for apparent inconsistencies thereby obtaining the sequences encoding the full-length protein.

Example 2

Identification of Single Nucleotide Polymorphisms in GPCRX Nucleic Acid Sequences

[0298] Variant sequences are also included in this application. A variant sequence can include a single nucleotide polymorphism (SNP). A SNP can, in some instances, be referred to as a “cSNP” to denote that the nucleotide sequence containing the SNP originates as a cDNA. A SNP can arise in several ways. For example, a SNP may be due to a substitution of one nucleotide for another at the polymorphic site. Such a substitution can be either a transition or a transversion. A SNP can also arise from a deletion of a nucleotide or an insertion of a nucleotide, relative to a reference allele. In this case, the polymorphic site is a site at which one allele bears a gap with respect to a particular nucleotide in another allele. SNPs occurring within genes may result in an alteration of the amino acid encoded by the gene at the position of the SNP. Intragenic SNPs may also be silent, when a codon including a SNP encodes the same amino acid as a result of the redundancy of the genetic code. SNPs occurring outside the region of a gene, or in an intron within a gene, do not result in changes in any amino acid sequence of a protein but may result in altered regulation of the expression pattern. Examples include alteration in temporal expression, physiological response regulation, cell type expression regulation, intensity of expression, and stability of transcribed message.

[0299] SeqCalling assemblies produced by the exon linking process were selected and extended using the following criteria. Genomic clones having regions with 98% identity to all or part of the initial or extended sequence were identified by BLASTN searches using the relevant sequence to query human genomic databases. The genomic clones that resulted were selected for further analysis because this identity indicates that these clones contain the genomic locus for these SeqCalling assemblies. These sequences were analyzed for putative coding regions as well as for similarity to the known DNA and protein sequences. Programs used for these analyses include Grail, Genscan, BLAST, HMMER, FASTA, Hybrid and other relevant programs.

[0300] Some additional genomic regions may have also been identified because selected SeqCalling assemblies map to those regions. Such SeqCalling sequences may have overlapped with regions defined by homology or exon prediction. They may also be included because the location of the fragment was in the vicinity of genomic regions identified by similarity or exon prediction that had been included in the original predicted sequence. The sequence so identified was manually assembled and then may have been extended using one or more additional sequences taken from CuraGen Corporation's human SeqCalling database. SeqCalling fragments suitable for inclusion were identified by the CuraTools™ program SeqExtend or by identifying SeqCalling fragments mapping to the appropriate regions of the genomic clones analyzed.

[0301] The regions defined by the procedures described above were then manually integrated and corrected for apparent inconsistencies that may have arisen, for example, from miscalled bases in the original fragments or from discrepancies between predicted exon junctions, EST locations and regions of sequence similarity, to derive the final sequence disclosed herein. When necessary, the process to identify and analyze SeqCalling assemblies and genomic clones was reiterated to derive the full length sequence (Alderborn et al., Determination of Single Nucleotide Polymorphisms by Real-time Pyrophosphate DNA Sequencing. Genome Research. 10 (8) 1249-1265, 2000).

Example 3

Quantitative Expression Analysis of Clones in Various Cells and Tissues

[0302] The quantitative expression of various clones was assessed using microtiter plates containing RNA samples from a variety of normal and pathology-derived cells, cell lines and tissues using real time quantitative PCR (RTQ PCR). RTQ PCR was performed on an Applied Biosystems ABI PRISM® 7700 or an ABI PRISM® 7900 HT Sequence Detection System. Various collections of samples are assembled on the plates, and referred to as Panel 1 (containing normal tissues and cancer cell lines), Panel 2 (containing samples derived from tissues from normal and cancer sources), Panel 3 (containing cancer cell lines), Panel 4 (containing cells and cell lines from normal tissues and cells related to inflammatory conditions), Panel 5D/5I (containing human tissues and cell lines with an emphasis on metabolic diseases), AI_comprehensive_panel (containing normal tissue and samples from autoimmune diseases), Panel CNSD.01 (containing central nervous system samples from normal and diseased brains) and CNS_neurodegeneration_panel (containing samples from normal and Alzheimer's diseased brains).

[0303] RNA integrity from all samples is controlled for quality by visual assessment of agarose gel electropherograms using 28S and 18S ribosomal RNA staining intensity ratio as a guide (2:1 to 2.5:1 28s: 18s) and the absence of low molecular weight RNAs that would be indicative of degradation products. Samples are controlled against genomic DNA contamination by RTQ PCR reactions run in the absence of reverse transcriptase using probe and primer sets designed to amplify across the span of a single exon.

[0304] First, the RNA samples were normalized to reference nucleic acids such as constitutively expressed genes (for example, β-actin and GAPDH). Normalized RNA (5 ul) was converted to cDNA and analyzed by RTQ-PCR using One Step RT-PCR Master Mix Reagents (Applied Biosystems; Catalog No. 4309169) and gene-specific primers according to the manufacturer's instructions.

[0305] In other cases, non-normalized RNA samples were converted to single strand cDNA (sscDNA) using Superscript II (Invitrogen Corporation; Catalog No. 18064-147) and random hexamers according to the manufacturer's instructions. Reactions containing up to 10 μg of total RNA were performed in a volume of 20 μl and incubated for 60 minutes at 42° C. This reaction can be scaled up to 50 μg of total RNA in a final volume of 100 μl. sscDNA samples are then normalized to reference nucleic acids as described previously, using 1× TaqMan® Universal Master mix (Applied Biosystems; catalog No. 4324020), following the manufacturer's instructions.

[0306] Probes and primers were designed for each assay according to Applied Biosystems Primer Express Software package (version I for Apple Computer's Macintosh Power PC) or a similar algorithm using the target sequence as input. Default settings were used for reaction conditions and the following parameters were set before selecting primers: primer concentration=250 nM, primer melting temperature (Tm) range=58°-60° C., primer optimal Tm=59° C., maximum primer difference=2° C., probe does not have 5′G, probe Tm must be 10° C. greater than primer Tm, amplicon size 75 bp to 100 bp. The probes and primers selected (see below) were synthesized by Synthegen (Houston, Tex., USA). Probes were double purified by HPLC to remove uncoupled dye and evaluated by mass spectroscopy to verify coupling of reporter and quencher dyes to the 5′ and 3′ ends of the probe, respectively. Their final concentrations were: forward and reverse primers, 900 nM each, and probe, 200 nM.

[0307] PCR conditions: When working with RNA samples, normalized RNA from each tissue and each cell line was spotted in each well of either a 96 well or a 384-well PCR plate (Applied Biosystems). PCR cocktails included either a single gene specific probe and primers set, or two multiplexed probe and primers sets (a set specific for the target clone and another gene-specific set multiplexed with the target probe). PCR reactions were set up using TaqMan® One-Step RT-PCR Master Mix (Applied Biosystems, Catalog No. 4313803) following manufacturer's instructions. Reverse transcription was performed at 48° C. for 30 minutes followed by amplification/PCR cycles as follows: 95° C. 10 min, then 40 cycles of 95° C. for 15 seconds, 60° C. for 1 minute. Results were recorded as CT values (cycle at which a given sample crosses a threshold level of fluorescence) using a log scale, with the difference in RNA concentration between a given sample and the sample with the lowest CT value being represented as 2 to the power of delta CT. The percent relative expression is then obtained by taking the reciprocal of this RNA difference and multiplying by 100.

[0308] When working with sscDNA samples, normalized sscDNA was used as described previously for RNA samples. PCR reactions containing one or two sets of probe and primers were set up as described previously, using 1× TaqMan® Universal Master mix (Applied Biosystems; catalog No. 4324020), following the manufacturer's instructions. PCR amplification was performed as follows: 95° C. 10 min, then 40 cycles of 95° C. for 15 seconds, 60° C. for 1 minute. Results were analyzed and processed as described previously.

[0309] Panels 1, 1.1, 1.2, and 1.3D

[0310] The plates for Panels 1, 1. 1, 1.2 and 1.3D include 2 control wells (genomic DNA control and chemistry control) and 94 wells containing cDNA from various samples. The samples in these panels are broken into 2 classes: samples derived from cultured cell lines and samples derived from primary normal tissues. The cell lines are derived from cancers of the following types: lung cancer, breast cancer, melanoma, colon cancer, prostate cancer, CNS cancer, squamous cell carcinoma, ovarian cancer, liver cancer, renal cancer, gastric cancer and pancreatic cancer. Cell lines used in these panels are widely available through the American Type Culture Collection (ATCC), a repository for cultured cell lines, and were cultured using the conditions recommended by the ATCC. The normal tissues found on these panels are comprised of samples derived from all major organ systems from single adult individuals or fetuses. These samples are derived from the following organs: adult skeletal muscle, fetal skeletal muscle, adult heart, fetal heart, adult kidney, fetal kidney, adult liver, fetal liver, adult lung, fetal lung, various regions of the brain, the spleen, bone marrow, lymph node, pancreas, salivary gland, pituitary gland, adrenal gland, spinal cord, thymus, stomach, small intestine, colon, bladder, trachea, breast, ovary, uterus, placenta, prostate, testis and adipose.

[0311] In the results for Panels 1, 1.1, 1.2 and 1.3D, the following abbreviations are used:

[0312] ca.=carcinoma,

[0313] *=established from metastasis,

[0314] met=metastasis,

[0315] s cell var=small cell variant,

[0316] non-s=non-sm=non-small,

[0317] squam=squamous,

[0318] pl. eff=pl effusion=pleural effusion,

[0319] glio=glioma,

[0320] astro=astrocytoma, and

[0321] neuro=neuroblastoma.

[0322] General_screening_panel_v1.4

[0323] The plates for Panel 1.4 include 2 control wells (genomic DNA control and chemistry control) and 94 wells containing cDNA from various samples. The samples in Panel 1.4 are broken into 2 classes: samples derived from cultured cell lines and samples derived from primary normal tissues. The cell lines are derived from cancers of the following types: lung cancer, breast cancer, melanoma, colon cancer, prostate cancer, CNS cancer, squamous cell carcinoma, ovarian cancer, liver cancer, renal cancer, gastric cancer and pancreatic cancer. Cell lines used in Panel 1.4 are widely available through the American Type Culture Collection (ATCC), a repository for cultured cell lines, and were cultured using the conditions recommended by the ATCC. The normal tissues found on Panel 1.4 are comprised of pools of samples derived from all major organ systems from 2 to 5 different adult individuals or fetuses. These samples are derived from the following organs: adult skeletal muscle, fetal skeletal muscle, adult heart, fetal heart, adult kidney, fetal kidney, adult liver, fetal liver, adult lung, fetal lung, various regions of the brain, the spleen, bone marrow, lymph node, pancreas, salivary gland, pituitary gland, adrenal gland, spinal cord, thymus, stomach, small intestine, colon, bladder, trachea, breast, ovary, uterus, placenta, prostate, testis and adipose. Abbreviations are as described for Panels 1, 1.1, 1.2, and 1.3D.

[0324] Panels 2D and 2.2

[0325] The plates for Panels 2D and 2.2 generally include 2 control wells and 94 test samples composed of RNA or cDNA isolated from human tissue procured by surgeons working in close cooperation with the Natlonal Cancer Institute's Cooperative Human Tissue Network (CHTN) or the Natlonal Disease Research Initiative (NDRI). The tissues are derived from human malignancies and in cases where indicated many malignant tissues have “matched margins” obtained from noncancerous tissue just adjacent to the tumor. These are termed normal adjacent tissues and are denoted “NAT” in the results below. The tumor tissue and the “matched margins” are evaluated by two independent pathologists (the surgical pathologists and again by a pathologist at NDRI or CHTN). This analysis provides a gross histopathological assessment of tumor differentiation grade. Moreover, most samples include the original surgical pathology report that provides information regamiding the clinical stage of the patient. These matched margins are taken from the tissue surrounding (i.e. immediately proximal) to the zone of surgery (designated “NAT”, for normal adjacent tissue, in Table RR). In addition, RNA and cDNA samples were obtained from various human tissues derived from autopsies performed on elderly people or sudden death victims (accidents, etc.). These tissues were ascertained to be free of disease and were purchased from various commercial sources such as Clontech (Palo Alto, Calif.), Research Genetics, and Invitrogen.

[0326] Panel 3D

[0327] The plates of Panel 3D are comprised of 94 cDNA samples and two control samples. Specifically, 92 of these samples are derived from cultured human cancer cell lines, 2 samples of human primary cerebellar tissue and 2 controls. The human cell lines are generally obtained from ATCC (American Type Culture Collection), NCI or the German tumor cell bank and fall into the following tissue groups: Squamous cell carcinoma of the tongue, breast cancer, prostate cancer, melanoma, epidermoid carcinoma, sarcomas, bladder carcinomas, pancreatic cancers, kidney cancers, leukemias/lymphomas, ovarian/uterine/cervical, gastric, colon, lung and CNS cancer cell lines. In addition, there are two independent samples of cerebellum. These cells are all cultured under standard recommended conditions and RNA extracted using the standard procedures. The cell lines in panel 3D and 1.3D are of the most common cell lines used in the scientific literature.

[0328] Panels 4D, 4R, and 4.1D

[0329] Panel 4 includes samples on a 96 well plate (2 control wells, 94 test samples) composed of RNA (Panel 4R) or cDNA (Panels 4D/4.D) isolated from various human cell lines or tissues related to inflammatory conditions. Total RNA from control normal tissues such as colon and lung (Stratagene, La Jolla, Calif.) and thymus and kidney (Clontech) was employed. Total RNA from liver tissue from cirrhosis patients and kidney from lupus patients was obtained from BioChain (Biochain Institute, Inc., Hayward, Calif.). Intestinal tissue for RNA preparation from patients diagnosed as having Crohn's disease and ulcerative colitis was obtained from the National Disease Research Interchange (NDRI) (Philadelphia, Pa.).

[0330] Astrocytes, lung fibroblasts, dermal fibroblasts, coronary artery smooth muscle cells, small airway epithelium, bronchial epithelium, microvascular dermal endothelial cells, microvascular lung endothelial cells, human pulmonary aortic endothelial cells, human umbilical vein endothelial cells were all purchased from Clonetics (Walkersville, Md.) and grown in the media supplied for these cell types by Clonetics. These primary cell types were activated with various cytokines or combinations of cytokines for 6 and/or 12-14 hours, as indicated. The following cytokines were used; IL-1 beta at approximately 1-5 ng/ml, TNF alpha at approximately 5-10 ng/ml, IFN gamma at approximately 20-50 ng/ml, IL-4 at approximately 5-10 ng/ml, IL-9 at approximately 5-10 ng/ml, IL-13 at approximately 5-10 ng/ml. Endothelial cells were sometimes starved for various times by culture in the basal media from Clonetics with 0.1% serum.

[0331] Mononuclear cells were prepared from blood of employees at CuraGen Corporation, using Ficoll. LAK cells were prepared from these cells by culture in DMEM 5% FCS (Hyclone), 100 μM non essential amino acids (Gibco/Life Technologies, Rockville, Md.), 1 mM sodium pyruvate (Gibco), mercaptoethanol 5.5×10−5M (Gibco), and 10 mM Hepes (Gibco) and Interleukin 2 for 4-6 days. Cells were then either activated with 10-20 ng/ml PMA and 1-2 μg/ml ionomycin, IL-12 at 5-10 ng/ml, IFN gamma at 20-50 ng/ml and IL-18 at 5-10 ng/ml for 6 hours. In some cases, mononuclear cells were cultured for 4-5 days in DMEM 5% FCS (Hyclone), 100 μM non essential amino acids (Gibco), 1 mM sodium pyruvate (Gibco), mercaptoethanol 5.5×10−5M (Gibco), and 10 mM Hepes (Gibco) with PHA (phytohemagglutinin) or PWM (pokeweed mitogen) at approximately 5 μg/ml. Samples were taken at 24, 48 and 72 hours for RNA preparation. MLR (mixed lymphocyte reaction) samples were obtained by taking blood from two donors, isolating the mononuclear cells using Ficoll and mixing the isolated mononuclear cells 1:1 at a final concentration of approximately 2×106cells/ml in DMEM 5% FCS (Hyclone), 100 M non essential amino acids (Gibco), 1 mM sodium pyruvate (Gibco), mercaptoethanol (5.5×10−5M) (Gibco), and 10 mM Hepes (Gibco). The MLR was cultured and samples taken at various time points ranging from 1-7 days for RNA preparation.

[0332] Monocytes were isolated from mononuclear cells using CD14 Miltenyi Beads, +ve VS selection columns and a Vario Magnet according to the manufacturer's instructions. Monocytes were differentiated into dendritic cells by culture in DMEM 5% fetal calf serum (FCS) (Hyclone, Logan, Utah), 100 μM non essential amino acids (Gibco), 1 mM sodium pyruvate (Gibco), mercaptoethanol 5.5×10−5M (Gibco), and 10 mM Hepes (Gibco), 50 ng/ml GMCSF and 5 ng/ml IL-4 for 5-7 days. Macrophages were prepared by culture of monocytes for 5-7 days in DMEM 5% FCS (Hyclone), 100 M non essential amino acids (Gibco), 1 mM sodium pyruvate (Gibco), mercaptoethanol 5.5×105M (Gibco), 10 mM Hepes (Gibco) and 10% AB Human Serum or MCSF at approximately 50 ng/ml. Monocytes, macrophages and dendritic cells were stimulated for 6 and 12-14 hours with lipopolysaccharide (LPS) at 100 ng/ml. Dendritic cells were also stimulated with anti-CD40 monoclonal antibody (Pharmingen) at 101 g/ml for 6 and 12-14 hours.

[0333] CD4 lymphocytes, CD8 lymphocytes and NK cells were also isolated from mononuclear cells using CD4, CD8 and CD56 Miltenyi beads, positive VS selection columns and a Vario Magnet according to the manufacturer's instructions. CD45RA and CD45RO CD4 lymphocytes were isolated by depleting mononuclear cells of CD8, CD56, CD 14 and CD19 cells using CD8, CD56, CD14 and CD19 Miltenyi beads and positive selection. CD45RO beads were then used to isolate the CD45RO CD4 lymphocytes with the remaining cells being CD45RA CD4 lymphocytes. CD45RA CD4, CD45RO CD4 and CD8 lymphocytes were placed in DMEM 5% FCS (Hyclone), 100 μM non essential amino acids (Gibco), 1 mM sodium pyruvate (Gibco), mercaptoethanol 5.5×10−5M (Gibco), and 10 mM Hepes (Gibco) and plated at 106cells/ml onto Falcon 6 well tissue culture plates that had been coated overnight with 0.5 μg/ml anti-CD28 (Pharmingen) and 3 ug/ml anti-CD3 (OKT3, ATCC) in PBS. After 6 and 24 hours, the cells were harvested for RNA preparation. To prepare chronically activated CD8 lymphocytes, we activated the isolated CD8 lymphocytes for 4 days on anti-CD28 and anti-CD3 coated plates and then harvested the cells and expanded them in DMEM 5% FCS (Hyclone), 100 μM non essential amino acids (Gibco), 1 mM sodium pyruvate (Gibco), mercaptoethanol 5.5×10−5M (Gibco), and 10 mM Hepes (Gibco) and IL-2. The expanded CD8 cells were then activated again with plate bound anti-CD3 and anti-CD28 for 4 days and expanded as before. RNA was isolated 6 and 24 hours after the second activation and after 4 days of the second expansion culture. The isolated NK cells were cultured in DMEM 5% FCS (Hyclone), 100 μM non essential amino acids (Gibco), 1 mM sodium pyruvate (Gibco), mercaptoethanol 5.5×10−5M (Gibco), and 10 mM Hepes (Gibco) and IL-2 for 4-6 days before RNA was prepared.

[0334] To obtain B cells, tonsils were procured from NDRI. The tonsil was cut up with sterile dissecting scissors and then passed through a sieve. Tonsil cells were then spun down and resupended at 106cells/ml in DMEM 5% FCS (Hyclone), 100 M non essential amino acids (Gibco), 1 mM sodium pyruvate (Gibco), mercaptoethanol 5.5×10−5M (Gibco), and 10 mM Hepes (Gibco). To activate the cells, we used PWM at 5 μg/ml or anti-CD40 (Pharmingen) at approximately 10 μg/ml and IL-4 at 5-10 ng/ml. Cells were harvested for RNA preparation at 24, 48 and 72 hours.

[0335] To prepare the primary and secondary Th1/Th2 and Tr1 cells, six-well Falcon plates were coated overnight with 10 μg/ml anti-CD28 (Pharmingen) and 2 μg/ml OKT3 (ATCC), and then washed twice with PBS. Umbilical cord blood CD4 lymphocytes (Poietic Systems, German Town, Md.) were cultured at 105-106cells/ml in DMEM 5% FCS (Hyclone), 100 μM non essential amino acids (Gibco), 1 mM sodium pyruvate (Gibco), mercaptoethanol 5.5×10−5M (Gibco), 10 mM Hepes (Gibco) and IL-2 (4 ng/ml). IL-12 (5 ng/ml) and anti-IL4 (1 μg/ml) were used to direct to Th1, while IL-4 (5 ng/ml) and anti-IFN gamma (1 μg/ml) were used to direct to Th2 and IL-10 at 5 ng/ml was used to direct to Tr1. After 4-5 days, the activated Th1, Th2 and Tr1 lymphocytes were washed once in DMEM and expanded for 4-7 days in DMEM 5% FCS (Hyclone), 100 μM non essential amino acids (Gibco), 1 mM sodium pyruvate (Gibco), mercaptoethanol 5.5×10−5M (Gibco), 10 mM Hepes (Gibco) and IL-2 (1 ng/ml). Following this, the activated Th1, Th2 and Tr1 lymphocytes were re-stimulated for 5 days with anti-CD28/OKT3 and cytokines as described above, but with the addition of anti-CD95L (1 μg/ml) to prevent apoptosis. After 4-5 days, the Th1, Th2 and Tr1 lymphocytes were washed and then expanded again with IL-2 for 4-7 days. Activated Th1 and Th2 lymphocytes were maintained in this way for a maximum of three cycles. RNA was prepared from primary and secondary Th1, Th2 and Tr1 after 6 and 24 hours following the second and third activations with plate bound anti-CD3 and anti-CD28 mAbs and 4 days into the second and third expansion cultures in Interleukin 2.

[0336] The following leukocyte cells lines were obtained from the ATCC: Ramos, EOL-1, KU-812. EOL cells were further differentiated by culture in 0.1 mM dbcAMP at 5×105cells/ml for 8 days, changing the media every 3 days and adjusting the cell concentration to 5×105cells/ml. For the culture of these cells, we used DMEM or RPMI (as recommended by the ATCC), with the addition of 5% FCS (Hyclone), 100 μM non essential amino acids (Gibco), 1 mM sodium pyruvate (Gibco), mercaptoethanol 5.5×10−5M (Gibco), 10 mM Hepes (Gibco). RNA was either prepared from resting cells or cells activated with PMA at 10 ng/ml and ionomycin at 1 μg/ml for 6 and 14 hours. Keratinocyte line CCD106 and an airway epithelial tumor line NCI-H292 were also obtained from the ATCC. Both were cultured in DMEM 5% FCS (Hyclone), 100 μM non essential amino acids (Gibco), 1 mM sodium pyruvate (Gibco), mercaptoethanol 5.5×10−5M (Gibco), and 10 mM Hepes (Gibco). CCD1106 cells were activated for 6 and 14 hours with approximately 5 ng/ml TNF alpha and 1 ng/ml IL-1 beta, while NCI-H292 cells were activated for 6 and 14 hours with the following cytokines: 5 ng/ml IL-4, 5 ng/ml IL-9, 5 ng/ml IL-13 and 25 ng/ml IFN gamma.

[0337] For these cell lines and blood cells, RNA was prepared by lysing approximately 107cells/ml using Trizol (Gibco BRL). Briefly, {fraction (1/10)} volume of bromochloropropane (Molecular Research Corporation) was added to the RNA sample, vortexed and after 10 minutes at room temperature, the tubes were spun at 14,000 rpm in a Sorvall SS34 rotor. The aqueous phase was removed and placed in a 15 ml Falcon Tube. An equal volume of isopropanol was added and left at −20° C. overnight. The precipitated RNA was spun down at 9,000 rpm for 15 min in a Sorvall SS34 rotor and washed in 70% ethanol. The pellet was redissolved in 300 μl of RNAse-free water and 35 μl buffer (Promega) 5 μl DTT, 7 μl RNAsin and 8 μl DNAse were added. The tube was incubated at 37° C. for 30 minutes to remove contaminating genomic DNA, extracted once with phenol chloroform and re-precipitated with {fraction (1/10)} volume of 3M sodium acetate and 2 volumes of 100% ethanol. The RNA was spun down and placed in RNAse free water. RNA was stored at −80° C.

[0338] AI_comprehensive Panel_v1.0

[0339] The plates for AI_comprehensive panel_v1.0 include two control wells and 89 test samples comprised of cDNA isolated from surgical and postmortem human tissues obtained from the Backus Hospital and Clinomics (Frederick, Md.). Total RNA was extracted from tissue samples from the Backus Hospital in the Facility at CuraGen. Total RNA from other tissues was obtained from Clinomics.

[0340] Joint tissues including synovial fluid, synovium, bone and cartilage were obtained from patients undergoing total knee or hip replacement surgery at the Backus Hospital. Tissue samples were immediately snap frozen in liquid nitrogen to ensure that isolated RNA was of optimal quality and not degraded. Additional samples of osteoarthritis and rheumatoid arthritis joint tissues were obtained from Clinomics. Normal control tissues were supplied by Clinomics and were obtained during autopsy of trauma victims.

[0341] Surgical specimens of psoriatic tissues and adjacent matched tissues were provided as total RNA by Clinomics. Two male and two female patients were selected between the ages of 25 and 47. None of the patients were taking prescription drugs at the time samples were isolated.

[0342] Surgical specimens of diseased colon from patients with ulcerative colitis and Crohns disease and adjacent matched tissues were obtained from Clinomics. Bowel tissue from three female and three male Crohn's patients between the ages of 41-69 were used. Two patients were not on prescription medication while the others were taking dexamethasone, phenobarbital, or tylenol. Ulcerative colitis tissue was from three male and four female patients. Four of the patients were taking lebvid and two were on phenobarbital.

[0343] Total RNA from post mortem lung tissue from trauma victims with no disease or with emphysema, asthma or COPD was purchased from Clinomics. Emphysema patients ranged in age from 40-70 and all were smokers, this age range was chosen to focus on patients with cigarette-linked emphysema and to avoid those patients with alpha-1 anti-trypsin deficiencies. Asthma patients ranged in age from 36-75, and excluded smokers to prevent those patients that could also have COPD. COPD patients ranged in age from 35-80 and included both smokers and non-smokers. Most patients were taking corticosteroids, and bronchodilators.

[0344] In the labels employed to identify tissues in the AI_comprehensive panel_v1.0 panel, the following abbreviations are used:

[0345] AI=Autoimmunity

[0346] Syn=Synovial

[0347] Normal=No apparent disease

[0348] Rep22/Rep20=individual patients

[0349] RA=Rheumatoid arthritis

[0350] Backus=From Backus Hospital

[0351] OA=Osteoarthritis

[0352] (SS) (BA) (MF)=Individual patients

[0353] Adj=Adjacent tissue

[0354] Match control=adjacent tissues

[0355] -M=Male

[0356] -F=Female

[0357] COPD=Chronic obstructive pulmonary disease

[0358] Panels 5D and 5I

[0359] The plates for Panel 5D and 5I include two control wells and a variety of cDNAs isolated from human tissues and cell lines with an emphasis on metabolic diseases. Metabolic tissues were obtained from patients enrolled in the Gestational Diabetes study. Cells were obtained during different stages in the differentiation of adipocytes from human mesenchymal stem cells. Human pancreatic islets were also obtained.

[0360] In the Gestational Diabetes study subjects are young (18-40 years), otherwise healthy women with and without gestational diabetes undergoing routine (elective) Caesarean section.

[0361] After delivery of the infant, when the surgical incisions were being repaired/closed, the obstetrician removed a small sample.

[0362] Patient 2: Diabetic Hispanic, overweight, not on insulin

[0363] Patient 7-9: Nondiabetic Caucasian and obese (BMI>30)

[0364] Patient 10: Diabetic Hispanic, overweight, on insulin

[0365] Patient 11: Nondiabetic African American and overweight

[0366] Patient 12: Diabetic Hispanic on insulin

[0367] Adipocyte differentiation was induced in donor progenitor cells obtained from Osirus (a division of Clonetics/BioWhittaker) in triplicate, except for Donor 3U which had only two replicates. Scientists at Clonetics isolated, grew and differentiated human mesenchymal stem cells (HuMSCs) for CuraGen based on the published protocol found in Mark F. Pittenger, et al., Multilineage Potential of Adult Human Mesenchymal Stem Cells Science Apr. 2, 1999: 143-147. Clonetics provided Trizol lysates or frozen pellets suitable for mRNA isolation and ds cDNA production. A general description of each donor is as follows:

[0368] Donor 2 and 3 U: Mesenchymal Stem cells, Undifferentiated Adipose

[0369] Donor 2 and 3 AM: Adipose, AdiposeMidway Differentiated

[0370] Donor 2 and 3 AD: Adipose, Adipose Differentiated

[0371] Human cell lines were generally obtained from ATCC (American Type Culture Collection), NCI or the German tumor cell bank and fall into the following tissue groups: kidney proximal convoluted tubule, uterine smooth muscle cells, small intestine, liver HepG2 cancer cells, heart primary stromal cells, and adrenal cortical adenoma cells. These cells are all cultured under standard recommended conditions and RNA extracted using the standard procedures. All samples were processed at CuraGen to produce single stranded cDNA.

[0372] Panel 5I contains all samples previously described with the addition of pancreatic islets from a 58 year old female patient obtained from the Diabetes Research Institute at the University of Miami School of Medicine. Islet tissue was processed to total RNA at an outside source and delivered to CuraGen for addition to panel 5I.

[0373] In the labels employed to identify tissues in the 5D and 5I panels, the following abbreviations are used:

[0374] GO Adipose=Greater Omentum Adipose

[0375] SK=Skeletal Muscle

[0376] UT=Uterus

[0377] PL=Placenta

[0378] AD=Adipose Differentiated

[0379] AM=Adipose Midway Differentiated

[0380] U=Undifferentiated Stem Cells

[0381] Panel CNSD.01

[0382] The plates for Panel CNSD.01 include two control wells and 94 test samples comprised of cDNA isolated from postmortem human brain tissue obtained from the Harvard Brain Tissue Resource Center. Brains are removed from calvaria of donors between 4 and 24 hours after death, sectioned by neuroanatomists, and frozen at −80° C. in liquid nitrogen vapor. All brains are sectioned and examined by neuropathologists to confirm diagnoses with clear associated neuropathology.

[0383] Disease diagnoses are taken from patient records. The panel contains two brains from each of the following diagnoses: Alzheimer's disease, Parkinson's disease, Huntington's disease, Progressive Supemuclear Palsy, Depression, and “Normal controls”. Within each of these brains, the following regions are represented: cingulate gyrus, temporal pole, globus palladus, substantia nigra, Brodman Area 4 (primary motor strip), Brodman Area 7 (parietal cortex), Brodman Area 9 (prefrontal cortex), and Brodman area 17 (occipital cortex). Not all brain regions are represented in all cases; e.g., Huntington's disease is characterized in part by neurodegeneration in the globus palladus, thus this region is impossible to obtain from confirmed Huntington's cases. Likewise Parkinson's disease is characterized by degeneration of the substantia nigra making this region more difficult to obtain. Normal control brains were examined for neuropathology and found to be free of any pathology consistent with neurodegeneration.

[0384] In the labels employed to identify tissues in the CNS panel, the following abbreviations are used:

[0385] PSP=Progressive supranuclear palsy

[0386] Sub Nigra=Substantia nigra

[0387] Glob Palladus=Globus palladus

[0388] Temp Pole=Temporal pole

[0389] Cing Gyr=Cingulate gyrus

[0390] BA 4=Brodman Area 4

[0391] Panel CNS_Neurodegeneration_V1.0

[0392] The plates for Panel CNS_Neurodegeneration_V1.0 include two control wells and 47 test samples comprised of cDNA isolated from postmortem human brain tissue obtained from the Harvard Brain Tissue Resource Center (McLean Hospital) and the Human Brain and Spinal Fluid Resource Center (VA Greater Los Angeles Healthcare System). Brains are removed from calvaria of donors between 4 and 24 hours after death, sectioned by neuroanatomists, and frozen at −80° C. in liquid nitrogen vapor. All brains are sectioned and examined by neuropathologists to confirm diagnoses with clear associated neuropathology.

[0393] Disease diagnoses are taken from patient records. The panel contains six brains from Alzheimer's disease (AD) patients, and eight brains from “Normal controls” who showed no evidence of dementia prior to death. The eight normal control brains are divided into two categories: Controls with no dementia and no Alzheimer's like pathology (Controls) and controls with no dementia but evidence of severe Alzheimer's like pathology, (specifically senile plaque load rated as level 3 on a scale of 0-3; 0=no evidence of plaques, 3=severe AD senile plaque load). Within each of these brains, the following regions are represented: hippocampus, temporal cortex (Brodman Area 21), parietal cortex (Brodman area 7), and occipital cortex (Brodman area 17). These regions were chosen to encompass all levels of neurodegeneration in AD. The hippocampus is a region of early and severe neuronal loss in AD; the temporal cortex is known to show neurodegeneration in AD after the hippocampus; the parietal cortex shows moderate neuronal death in the late stages of the disease; the occipital cortex is spared in AD and therefore acts as a “control” region within AD patients. Not all brain regions are represented in all cases.

[0394] In the labels employed to identify tissues in the CNS_Neurodegeneration_V1.0 panel, the following abbreviations are used:

[0395] AD=Alzheimer's disease brain; patient was demented and showed AD-like pathology upon autopsy

[0396] Control=Control brains; patient not demented, showing no neuropathology

[0397] Control (Path)=Control brains; pateint not demented but showing sever AD-like pathology

[0398] SupTemporal Ctx=Superior Temporal Cortex

[0399] Inf Temporal Ctx=Inferior Temporal Cortex

[0400] A. CG142263-01/GMAP000916_Aand GMAP001524_A: Olfactory Receptor

[0401] Expression of gene CG142263-01 and variant GMAP001524_A was assessed using the primer-probe sets Ag2225 and Ag1826, described in Tables AA and AB. Results of the RTQ-PCR runs are shown in Tables AC and AD.

2TABLE AAProbe Name Ag2225StartSEQ IDPrimersSequencesLengthPositionNO:Forward5′-tctcagcaatctgtcactcatg-3′22187153ProbeTET-5′-tctgctactcctccgtcattacccct-3′-TAMRA26213154Reverse5′-gacacaaagttcaccagcatct-3′22240155

[0402]

3

TABLE AB

Probe Name Ag1826

Start
SEQ ID

Primers
Sequences
Length
Position
NO:

Forward
5′-tctcagcaatctgtcactcatg-3′
22
187
156

Probe
TET-5′-tctgctactcctccgtcattacccct-3′-TAMRA
26
213
157

Reverse
5′-gacacaaagttcaccagcatct-3′
22
240
158

[0403]

4

TABLE AC

Panel 1.3D

Rel. Exp. (%) Ag2225, Run

Rel. Exp. (%) Ag2225, Run

Tissue Name
165974841
Tissue Name
165974841

Liver adenocarcinoma
0.0
Kidney (fetal)
0.0

Pancreas
0.0
Renal ca. 786-0
0.0

Pancreatic ca. CAPAN 2
0.0
Renal ca. A498
0.0

Adrenal gland
0.0
Renal ca. RXF 393
0.0

Thyroid
0.0
Renal ca. ACHN
0.0

Salivary gland
0.0
Renal ca. UO-31
0.0

Pituitary gland
0.0
Renal ca. TK-10
0.0

Brain (fetal)
2.1
Liver
0.0

Brain (whole)
0.0
Liver (fetal)
0.0

Brain (amygdala)
0.0
Liver ca. (hepatoblast)
0.0

HepG2

Brain (cerebellum)
0.0
Lung
0.0

Brain (hippocampus)
0.0
Lung (fetal)
0.0

Brain (substantia nigra)
0.0
Lung ca. (small cell) LX-1
0.0

Brain (thalamus)
0.0
Lung ca. (small cell)
0.0

NCI-H69

Cerebral Cortex
0.0
Lung ca. (s.cell var.)
0.0

SHP-77

Spinal cord
0.0
Lung ca. (large cell)NCI-
0.0

H460

glio/astro U87-MG
0.0
Lung ca. (non-sm. cell)
8.2

A549

glio/astro U-118-MG
0.0
Lung ca. (non-s.cell)
0.0

NCI-H23

astrocytoma SW1783
0.0
Lung ca. (non-s.cell)
0.0

HOP-62

neuro*; met SK-N-AS
0.0
Lung ca. (non-s.cl) NCI-
0.0

H522

astrocytoma SF-539
0.0
Lung ca. (squam.) SW
0.0

900

astrocytoma SNB-75
0.0
Lung ca. (squam.) NCI-
0.0

H596

glioma SNB-19
0.0
Mammary gland
0.0

glioma U251
0.0
Breast ca.* (pl.ef) MCF-7
24.0

glioma SF-295
0.0
Breast ca.* (pl.ef) MDA-
17.8

MB-231

Heart (fetal)
0.0
Breast ca.* (pl.ef) T47D
29.3

Heart
0.0
Breast ca. BT-549
0.0

Skeletal muscle (fetal)
0.0
Breast ca. MDA-N
0.0

Skeletal muscle
0.0
Ovary
0.0

Bone marrow
0.0
Ovarian ca. OVCAR-3
0.0

Thymus
0.0
Ovarian ca. OVCAR-4
0.0

Spleen
0.0
Ovarian ca. OVCAR-5
0.0

Lymph node
0.0
Ovarian ca. OVCAR-8
6.3

Colorectal
6.4
Ovarian ca. IGROV-1
0.0

Stomach
0.0
Ovarian ca.* (ascites)
5.4

SK-OV-3

Small intestine
0.0
Uterus
0.0

Colon ca. SW480
0.0
Plancenta
0.0

Colon ca.* SW620(SW480
0.0
Prostate
0.0

met)

Colon ca. HT29
0.0
Prostate ca.* (bone
0.0

met)PC-3

Colon ca. HCT-116
0.0
Testis
0.0

Colon ca. CaCo-2
0.0
Melanoma Hs688(A).T
0.0

Colon ca.
0.0
Melanoma* (met)
0.0

tissue(ODO3866)

Hs688(B).T

Colon ca. HCC-2998
0.0
Melanoma UACC-62
100.0

Gastric ca.* (liver met)
0.0
Melanoma M14
0.0

NCI-N87

Bladder
0.0
Melanoma LOX IMVI
0.0

Trachea
0.0
Melanoma* (met) SK-
0.0

MEL-5

Kidney
0.0
Adipose
0.0

[0404]

5

TABLE AD

Panel 4D

Rel. Exp. (%)
Rel. Exp. (%)

Rel. Exp. (%)
Rel. Exp. (%)

Ag1826, Run
Ag2225, Run

Ag1826, Run
Ag2225, Run

Tissue Name
165809048
164019474
Tissue Name
165809048
164019474

Secondary Th1 act
0.0
0.0
HUVEC IL-1beta
0.0
0.0

Secondary Th2 act
2.9
0.0
HUVEC IFN gamma
0.0
0.0

Secondary Tr1 act
2.9
16.6
HUVEC TNF alpha +
0.0
0.0

IFN gamma

Secondary Th1 rest
0.0
0.0
HUVEC TNF alpha +
0.0
0.0

IL4

Secondary Th2 rest
0.0
0.0
HUVEC IL-11
0.0
0.0

Secondary Tr1 rest
0.0
0.0
Lung Microvascular
0.0
0.0

EC none

Primary Th1 act
0.0
0.0
Lung Microvascular
0.0
0.0

EC TNF alpha + IL-

1beta

Primary Th2 act
0.0
0.0
Microvascular
0.0
0.0

Dermal EC none

Primary Tr1 act
0.0
0.0
Microsvasular Dermal
0.0
0.0

EC TNF alpha + IL-

1beta

Primary Th1 rest
0.0
0.0
Bronchial epithelium
0.0
0.0

TNF alpha + IL1beta

Primary Th2 rest
0.0
0.0
Small airway
0.0
0.0

epithelium none

Primary Tr1 rest
0.0
0.0
Small airway
0.0
0.0

epithelium TNF alpha +

IL-1beta

CD45RA CD4
0.0
0.0
Coronery artery SMC
0.0
0.0

lymphocyte act

rest

CD45RO CD4
0.0
0.0
Coronery artery SMC
0.0
0.0

lymphocyte act

TNF alpha + IL-1beta

CD8 lymphocyte act
0.0
0.0
Astrocytes rest
0.0
0.0

Secondary CD8
0.0
0.0
Astrocytes TNF alpha +
0.0
0.0

lymphocyte rest

IL-1beta

Secondary CD8
0.0
0.0
KU-812 (Basophil)
0.0
0.0

lymphocyte act

rest

CD4 lymphocyte
0.0
0.0
KU-812 (Basophil)
0.0
0.0

none

PMA/ionomycin

2ry
0.0
0.0
CCD1106
0.0
0.0

Th1/Th2/Tr1_anti-

(Keratinocytes) none

CD95 CH11

LAK cells rest
0.0
0.0
CCD1106
0.0
0.0

(Keratinocytes)

TNF alpha + IL-1beta

LAK cells IL-2
0.0
0.0
Liver cirrhosis
100.0
100.0

LAK cells IL-2 + IL-
0.0
0.0
Lupus kidney
2.7
0.0

12

LAK cells IL-2 + IFN
0.0
0.0
NCI-H292 none
0.0
0.0

gamma

LAK cells IL-2 + IL-
0.0
0.0
NCI-H292 IL-4
0.0
0.0

18

LAK cells
0.0
0.0
NCI-H292 IL-9
0.0
0.0

PMA/ionomycin

NK Cells IL-2 rest
0.0
0.0
NCI-H292 IL-13
0.0
0.0

Two Way MLR 3
0.0
0.0
NCI-H292 IFN
0.0
0.0

day

gamma

Two Way MLR 5
0.0
0.0
HPAEC none
0.0
0.0

day

Two Way MLR 7
0.0
0.0
HPAEC TNF alpha +
0.0
0.0

day

IL-1beta

PBMC rest
0.0
0.0
Lung fibroblast none
0.0
0.0

PBMC PWM
0.0
0.0
Lung fibroblast TNF
0.0
0.0

alpha + IL-1beta

PBMC PHA-L
0.0
0.0
Lung fibroblast IL-4
0.0
0.0

Ramos (B cell) none
0.0
0.0
Lung fibroblast IL-9
5.9
0.0

Ramos (B cell)
0.0
0.0
Lung fibroblast IL-13
0.0
0.0

ionomycin

B lymphocytes PWM
0.0
0.0
Lung fibroblast IFN
0.0
0.0

gamma

B lymphocytes
1.8
0.0
Dermal fibroblast
0.0
0.0

CD40L and IL-4

CCD1070 rest

EOL-1 dbcAMP
0.0
0.0
Dermal fibroblast
0.0
0.0

CCD1070 TNF alpha

EOL-1 dbcAMP
0.0
0.0
Dermal fibroblast
0.0
0.0

PMA/ionomycin

CCD1070 IL-1beta

Dendritic cells none
0.0
0.0
Dermal fibroblast IFN
0.0
0.0

gamma

Dendritic cells LPS
4.7
0.0
Dermal fibroblast IL-4
0.0
0.0

Dendritic cells anti-
13.1
0.0
IBD Colitis 2
28.9
91.4

CD40

Monocytes rest
0.0
0.0
IBD Crohn's
7.7
0.0

Monocytes LPS
2.0
0.0
Colon
23.3
83.5

Macrophages rest
4.9
30.4
Lung
3.0
0.0

Macrophages LPS
0.0
0.0
Thymus
0.0
0.0

HUVEC none
0.0
0.0
Kidney
0.0
0.0

HUVEC starved
0.0
0.0

[0405] Panel 1.3D Summary: Ag2225 Low but significant expression of the CG142263-01 gene is limited to a single melanoma cell line (CT=33.4). Therefore, expression of this gene may be used to distinguish melanoma cell lines from the other samples on this panel. Furthermore, therapeutic modulation of the activity of the GPCR encoded by this gene may be beneficial in the treatment or detection of melanoma.

[0406] Panel 2.2 Summary: Ag2225 Expression of the CG142263-01 gene low/undetectable (CTs>35) across all of the samples on this panel.

[0407] Panel 4D Summary: Ag1826/Ag2225 Low but significant levels of expression of the CG142263-01 gene is detected in antiCD40 stimulated dendritic cells as well as normal colon, inflammatory bowel disease (IBD) colitis, and liver cirrhosis. Dendritic cells represent an important specialized population of inflammatory cells that function as activators/co-stimulators of T cells, as well as possessing antigen presentation functions. Owing to their importance in inflammatory processes and inflammatory cascades, therapeutic modulation of this gene product may reduce or eliminate the symptoms of patients suffering from asthma, allergies, chronic obstructive pulmonary disease, emphysema, Crohn's disease, ulcerative colitis, rheumatoid arthritis, psoriasis, and osteoarthritis. The presence of this gene in liver cirrhosis (a component of which involves liver inflammation and fibrosis) suggests that therapeutic agents involving this gene may be useful in reducing or inhibiting the inflammation associated with fibrotic and inflammatory diseases. In addition, antibodies to this putative GPCR could also be used for the diagnosis of liver cirrhosis.

[0408] B. CG143433-01/GMAP000818_E: Olfactory Receptor

[0409] Expression of gene CG143433-01 was assessed using the primer-probe set Ag2220, described in Table BA. Results of the RTQ-PCR runs are shown in Table BB.

6TABLE BAProbe Name Ag2220StartSEQ IDPrimersSequencesLengthPositionNO:Forward5′-tgtaccatttcctcagcagtct-3′22179159ProbeTET-5′-tctgccagtcttctgtcattacccca-3′-TAMRA26215160Reverse5′-gacacaaaattcaccagcattt-3′22242161

[0410]

7

TABLE BB

Panel 1.3D

Rel. Exp. (%) Ag2220, Run

Rel. Exp. (%) Ag2220, Run

Tissue Name
165974840
Tissue Name
165974840

Liver adenocarcinoma
0.0
Kidney (fetal)
0.0

Pancreas
0.0
Renal ca. 786-0
0.0

Pancreatic ca. CAPAN 2
0.0
Renal ca. A498
0.0

Adrenal gland
0.0
Renal ca. RXF 393
0.0

Thyroid
0.0
Renal ca. ACHN
0.0

Salivary gland
0.0
Renal ca. UO-31
0.0

Pituitary gland
0.0
Renal ca. TK-10
0.0

Brain (fetal)
0.0
Liver
0.0

Brain (whole)
0.0
Liver (fetal)
0.0

Brain (amygdala)
0.0
Liver ca. (hepatoblast)
0.0

HepG2

Brain (cerebellum)
0.0
Lung
0.0

Brain (hippocampus)
0.0
Lung (fetal)
0.0

Brain (substantia nigra)
0.0
Lung ca. (small cell) LX-1
0.0

Brain (thalamus)
0.0
Lung ca. (small cell)
0.0

NCI-H69

Cerebral Cortex
0.0
Lung ca. (s.cell var.)
0.0

SHP-77

Spinal cord
0.0
Lung ca. (large cell)NCI-
0.0

H460

glio/astro U87-MG
0.0
Lung ca. (non-sm. cell)
0.0

A549

glio/astro U-118-MG
0.0
Lung ca. (non-s.cell)
0.0

NCI-H23

astrocytoma SW1783
0.0
Lung ca. (non-s.cell)
0.0

HOP-62

neuro*; met SK-N-AS
0.0
Lung ca. (non-s.cl) NCI-
0.0

H522

astrocytoma SF-539
0.0
Lung ca. (squam.) SW
0.0

900

astrocytoma SNB-75
0.0
Lung ca. (squam.) NCI-
0.0

H596

glioma SNB-19
0.0
Mammary gland
0.0

glioma U251
0.0
Breast ca.* (pl.ef) MCF-7
43.8

glioma SF-295
0.0
Breast ca.* (pl.ef) MDA-
0.0

MB-231

Heart (Fetal)
0.0
Breast ca.* (pl.ef) T47D
100.0

Heart
0.0
Breast ca. BT-549
0.0

Skeletal muscle (Fetal)
0.0
Breast ca. MDA-N
0.0

Skeletal muscle
0.0
Ovary
0.0

Bone marrow
0.0
Ovarian ca. OVCAR-3
0.0

Thymus
0.0
Ovarian ca. OVCAR-4
0.0

Spleen
0.0
Ovarian ca. OVCAR-5
0.0

Lymph node
0.0
Ovarian ca. OVCAR-8
0.0

Colorectal
0.0
Ovarian ca. IGROV-1
0.0

Stomach
0.0
Ovarian ca. (ascites) SK-
0.0

OV-3

Small intestine
0.0
Uterus
0.0

Colon ca. SW480
0.0
Placenta
22.2

Colon ca.* SW620
0.0
Prostate
0.0

(SW480 met)

Colon ca. HT29
0.0
Prostate ca.* (bone met)
7.9

PC-3

Colon ca. HCT-116
0.0
Testis
0.0

Colon ca. CaCo-2
0.0
Melanoma Hs688(A).T
0.0

CC Well to Mod Diff
0.0
Melanoma* (met)
0.0

(ODO3866)

Hs688(B).T

Colon ca. HCC-2998
0.0
Melanoma UACC-62
76.8

Gastric ca. (liver met)
0.0
Melanoma M14
0.0

NCI-N87

Bladder
0.0
Melanoma LOX IMVI
1.9

Trachea
0.0
Melanoma* (met) SK-
0.0

MEL-5

Kidney
0.0
Adipose
0.0

[0411] Panel 1.3D Summary: Ag2220 Expression of the CG143433-01 gene is low but significant in a single breast cancer cell line and a single melanoma cell line. Therefore, expression of this gene may be used to distinguish breast and melanoma cancer cell lines from the other samples on this panel. Furthermore, therapeutic modulation of the activity of the GPCR encoded by this gene may be beneficial in the treatment of breast cancer and melanoma.

[0412] Panel 2.2 Summary: Ag2220 Expression of the CG143433-01 gene is low/undetectable (CTs>35) across all of the samples on this panel (data not shown).

[0413] Panel 4D Summary: Ag2220 Expression of the CG143433-01 gene is low/undetectable (CTs>35) across all of the samples on this panel (data not shown).

[0414] C. GMAP000818_A—1/GMAP000818_C/CG55976-03 and CG55976-02 and CG55976-01/GMAP000818_F: Olfactory Receptor

[0415] Expression of gene GMAP000818_A—1, variant CG55976-01, and full length phyical clone CG55976-02 was assessed using the primer-probe sets Ag2354, Ag2211 and Ag2221, described in Tables CA, CB, and CC. Results of the RTQ-PCR runs are shown in Tables CD, CE and CF.

8TABLE CAProbe Name Ag2354StartSEQ IDPrimersSequencesLengthPositionNO:Forward5′-ttgtgatcttttccctctcttg-3′22537162ProbeTET-5′-ctcctgctccagcacctacatcaatg-3′-TAMRA26564163Reverse5′-gcactcaagaccagaaccagta-3′22593164

[0416]

9

TABLE CB

Probe Name Ag2211

Start
SEQ ID

Primers
Sequences
Length
Position
NO:

Forward
5′-ttgtgatcttttccctctcttg-3′
22
537
165

Probe
TET-5′-ctcctgctccagcacctacatcaatg-3′-TAMRA
26
564
166

Reverse
5′-gcactcaagaccagaaccagta-3′
22
593
167

[0417]

10

TABLE CC

Probe Name Ag2221

Start
SEQ ID

Primers
Sequences
Length
Position
NO:

Forward
5′-ttgtgatcttttccctctcttg-3′
22
537
168

Probe
TET-5′-ctcctgctccagcacctacatcaatg-3′-TAMRA
26
564
169

Reverse
5′-gcactcaagaccagaaccagta-3′
22
593
170

[0418]

11

TABLE CD

Panel 1.3D

Rel. Exp. (%)
Rel. Exp. (%)

Rel. Exp. (%)
Rel. Exp. (%)

Ag2211, Run
Ag2221, Run

Ag2211, Run
Ag2221, Run

Tissue Name
166003999
165974927
Tissue Name
166003999
165974927

Liver
0.0
0.0
Kidney (fetal)
0.0
0.0

adenocarcinoma

Pancreas
0.0
43.8
Renal ca. 786-0
0.0
5.9

Pancreatic ca.
0.0
0.0
Renal ca. A498
0.0
0.0

CAPAN 2

Adrenal gland
0.0
0.0
Renal ca. RXF
0.0
0.0

393

Thyroid
0.0
9.2
Renal ca. ACHN
0.0
0.0

Salivary gland
0.0
0.0
Renal ca. UO-31
0.0
0.0

Pituitary gland
0.0
0.0
Renal ca. TK-10
0.0
0.0

Brain (fetal)
0.0
0.0
Liver
7.2
0.0

Brain (whole)
0.0
0.0
Liver (fetal)
0.0
0.0

Brain (amygdala)
0.0
0.0
Liver ca.
0.0
0.0

(hepatoblast)

HepG2

Brain (cerebellum)
0.0
0.0
Lung
0.0
7.7

Brain (hippocampus)
0.0
0.0
Lung (fetal)
0.0
0.0

Brain (substantia
0.0
0.0
Lung ca. (small
0.0
0.0

nigra)

cell) LX-1

Brain (thalamus)
0.0
0.0
Lung ca. (small
0.0
0.0

cell) NCI-H69

Cerebral Cortex
0.0
0.0
Lung ca. (s.cell
3.3
6.9

var.) SHP-77

Spinal cord
0.0
0.0
Lung ca. (large
0.0
0.0

cell)NCI-H460

glio/astro U87-MG
0.0
0.0
Lung ca. (non-
0.0
0.0

sm. cell) A549

glio/astro U-118-
0.0
0.0
Lung ca. (non-
4.0
0.0

MG

s.cell) NCI-H23

astrocytoma
0.0
0.0
Lung ca. (non-
0.0
0.0

SW1783

s.cell) HOP-62

neuro*; met SK-N-
0.0
0.0
Lung ca. (non-
0.0
0.0

AS

s.cl) NCI-H522

astrocytoma SF-539
0.0
9.4
Lung ca.
0.0
0.0

(squam.) SW 900

astrocytoma SNB-75
0.0
0.0
Lung ca.
3.3
0.0

(squam.) NCI-

H596

glioma SNB-19
5.8
0.0
Mammary gland
0.0
0.0

glioma U251
0.0
0.0
Breast ca.*
52.9
100.0

(pl.ef) MCF-7

glioma SF-295
0.0
0.0
Breast ca.*
0.0
0.0

(pl.ef) MDA-

MB-231

Heart (fetal)
0.0
0.0
Breast ca.*
66.9
55.5

(pl.ef) T47D

Heart
0.0
0.0
Breast ca. BT-
0.0
0.0

549

Skeletal muscle
0.0
0.0
Breast ca. MDA-N
0.0
0.0

(fetal)

Skeletal muscle
0.0
0.0
Ovary
0.0
0.0

Bone marrow
0.0
0.0
Ovarian ca.
0.0
0.0

OVCAR-3

Thymus
0.0
0.0
Ovarian ca.
8.0
0.0

OVCAR-4

Spleen
0.0
0.0
Ovarian ca.
0.0
0.0

OVCAR-5

Lymph node
0.0
0.0
Ovarian ca.
0.0
0.0

OVCAR-8

Colorectal
23.7
4.9
Ovarian ca.
0.0
0.0

IGROV-1

Stomach
0.0
0.0
Ovarian ca.*
0.0
0.0

(ascites) SK-OV-3

Small intestine
0.0
0.0
Uterus
0.0
0.0

Colon ca. SW480
0.0
0.0
Plancenta
48.0
21.6

Colon ca.*
0.0
0.0
Prostate
0.0
0.0

SW620(SW480 met)

Colon ca. HT29
0.0
0.0
Prostate ca.*
25.9
35.1

(bone met)PC-3

Colon ca. HCT-116
0.0
8.4
Testis
34.6
26.8

Colon ca. CaCo-2
0.0
0.0
Melanoma
0.0
0.0

Hs688(A).T

Colon ca.
0.0
0.0
Melanoma*
0.0
0.0

tissue(ODO3866)

(met)

Hs688(B).T

Colon ca. HCC-2998
0.0
0.0
Melanoma
100.0
94.6

UACC-62

Gastric ca.* (liver
0.0
0.0
Melanoma M14
0.0
0.0

met) NCI-N87

Bladder
0.0
13.3
Melanoma LOX
0.0
0.0

IMVI

Trachea
0.0
0.0
Melanoma*
0.0
0.0

(met) SK-MEL-5

Kidney
0.0
10.9
Adipose
0.0
0.0

[0419]

12

TABLE CE

Panel 2.2

Rel. Exp. (%)
Rel. Exp. (%)

Rel. Exp. (%)
Rel. Exp. (%)

Ag2211, Run
Ag2221, Run

Ag2211, Run
Ag2221, Run

Tissue Name
174255642
174285452
Tissue Name
174255642
174285452

Normal Colon
0.0
0.0
Kidney Margin
0.0
0.0

(OD04348)

Colon cancer
17.8
0.0
Kidney malignant
0.0
2.3

(OD06064)

cancer

(OD06204B)

Colon Margin
0.0
0.0
Kidney normal
0.0
0.0

(OD06064)

adjacent tissue

(OD06204E)

Colon cancer
0.0
0.0
Kidney Cancer
0.0
0.0

(OD06159)

(OD04450-01)

Colon Margin
20.7
0.0
Kidney Margin
0.0
0.0

(OD06159)

(OD04450-03)

Colon cancer
0.0
0.0
Kidney Cancer
0.0
0.0

(OD06297-04)

8120613

Colon Margin
16.4
0.0
Kidney Margin
0.0
0.0

(OD06297-015)

8120614

CC Gr.2 ascend
0.0
0.0
Kidney Cancer
0.0
0.0

colon (ODO3921)

9010320

CC Margin
0.0
0.0
Kidney Margin
0.0
0.0

(ODO3921)

9010321

Colon cancer
0.0
6.8
Kidney Cancer
0.0
0.0

metastasis

8120607

(OD06104)

Lung Margin
0.0
0.0
Kidney Margin
0.0
0.0

(OD06104)

8120608

Colon mets to lung
17.6
0.0
Normal Uterus
0.0
0.0

(OD04451-01)

Lung Margin
0.0
0.0
Uterine Cancer
7.7
0.0

(OD04451-02)

064011

Normal Prostate
0.0
0.0
Normal Thyroid
0.0
0.0

Prostate Cancer
0.0
0.0
Thyroid Cancer
0.0
0.0

(OD04410)

064010

Prostate Margin
0.0
0.0
Thyroid Cancer
0.0
0.0

(OD04410)

A302152

Normal Ovary
0.0
0.0
Thyroid Margin
0.0
0.0

A302153

Ovarian cancer
0.0
0.0
Normal Breast
0.0
0.0

(OD06283-03)

Ovarian Margin
0.0
0.0
Breast Cancer
9.6
0.0

(OD06283-07)

(OD04566)

Ovarian Cancer
100.0
16.2
Breast Cancer 1024
0.0
0.0

064008

Ovarian cancer
0.0
0.0
Breast Cancer
0.0
0.0

(OD06145)

(OD04590-01)

Ovarian Margin
0.0
0.0
Breast Cancer Mets
0.0
0.0

(OD06145)

(OD04590-03)

Ovarian cancer
0.0
0.0
Breast Cancer
0.0
0.0

(OD06455-03)

Metastasis

(OD04655-05)

Ovarian Margin
0.0
0.0
Breast Cancer
37.1
6.8

(OD06455-07)

064006

Normal Lung
0.0
0.0
Breast Cancer
0.0
0.0

9100266

Invasive poor diff.
16.6
1.3
Breast Margin
0.0
0.0

lung adeno

9100265

(ODO4945-01)

Lung Margin
0.0
0.0
Breast Cancer
0.0
0.0

(ODO4945-03)

A209073

Lung Malignant
0.0
4.0
Breast Margin
0.0
3.1

Cancer (OD03126)

A2090734

Lung Margin
0.0
0.0
Breast cancer
32.1
2.7

(OD03126)

(OD06083)

Lung Cancer
0.0
0.0
Breast cancer node
0.0
0.0

(OD05014A)

metastasis

(OD06083)

Lung Margin
0.0
0.0
Normal Liver
0.0
0.0

(OD05014B)

Lung cancer
0.0
0.0
Liver Cancer 1026
0.0
0.0

(OD06081)

Lung Margin
0.0
0.0
Liver Cancer 1025
7.9
0.0

(OD06081)

Lung Cancer
0.0
1.8
Liver Cancer 6004-T
0.0
0.0

(OD04237-01)

Lung Margin
0.0
100.0
Liver Tissue 6004-N
0.0
0.0

(OD04237-02)

Ocular Melanoma
0.0
0.0
Liver Cancer 6005-T
0.0
0.0

Metastasis

Ocular Melanoma
0.0
0.0
Liver Tissue 6005-N
0.0
0.0

Margin (Liver)

Melanoma
16.4
6.4
Liver Cancer
18.0
6.5

Metastasis

064003

Melanoma Margin
0.0
0.0
Normal Bladder
4.6
0.0

(Lung)

Normal Kidney
0.0
0.0
Bladder Cancer
0.0
0.0

1023

Kidney Ca, Nuclear
0.0
0.0
Bladder Cancer
50.7
30.8

grade 2 (OD04338)

A302173

Kidney Margin
0.0
0.0
Normal Stomach
0.0
0.0

(OD04338)

Kidney Ca Nuclear
0.0
0.0
Gastric Cancer
0.0
0.0

grade 1/2

9060397

(OD04339)

Kidney Margin
66.9
0.0
Stomach Margin
0.0
0.0

(OD04339)

9060396

Kidney Ca, Clear
0.0
0.0
Gastric Cancer
0.0
6.1

cell type

9060395

(OD04340)

Kidney Margin
0.0
0.0
Stomach Margin
0.0
15.2

(OD04340)

9060394

Kidney Ca, Nuclear
0.0
0.0
Gastric Cancer
0.0
0.0

grade 3 (OD04348)

064005

[0420]

13

TABLE CF

Panel 4D

Rel. Exp. (%)
Rel. Exp. (%)

Rel. Exp. (%)
Rel. Exp. (%)

Ag2211, Run
Ag2221, Run

Ag2211, Run
Ag2221, Run

Tissue Name
163630872
163923865
Tissue Name
163630872
163923865

Secondary Th1 act
0.0
0.0
HUVEC IL-1beta
4.2
0.0

Secondary Th2 act
0.0
0.0
HUVEC IFN gamma
0.0
0.0

Secondary Tr1 act
0.0
3.6
HUVEC TNF alpha +
0.0
0.0

IFN gamma

Secondary Th1 rest
0.0
0.0
HUVEC TNF alpha +
0.0
0.0

IL4

Secondary Th2 rest
0.0
0.0
HUVEC IL-11
0.0
0.0

Secondary Tr1 rest
0.0
0.0
Lung Microvascular
5.8
0.0

EC none

Primary Th1 act
0.0
8.7
Lung Microvascular
0.0
0.0

EC TNF alpha + IL-

1beta

Primary Th2 act
0.0
0.0
Microvascular
0.0
0.0

Dermal EC none

Primary Tr1 act
0.0
11.3
Microsvasular Dermal
0.0
7.4

EC TNF alpha + IL-

1beta

Primary Th1 rest
0.0
0.0
Bronchial epithelium
0.0
0.0

TNF alpha + IL1beta

Primary Th2 rest
0.0
0.0
Small airway
0.0
0.0

epithelium none

Primary Tr1 rest
0.0
0.0
Small airway
0.0
0.0

epithelium TNF alpha +

IL-1beta

CD45RA CD4
0.0
0.0
Coronery artery SMC
0.0
0.0

lymphocyte act

rest

CD45RO CD4
0.0
0.0
Coronery artery SMC
0.0
0.0

lymphocyte act

TNF alpha + IL-1beta

CD8 lymphocyte act
0.0
0.0
Astrocytes rest
0.0
0.0

Secondary CD8
0.0
0.0
Astrocytes TNF alpha +
0.0
0.0

lymphocyte rest

IL-1beta

Secondary CD8
0.0
0.0
KU-812 (Basophil)
3.1
0.0

lymphocyte act

rest

CD4 lymphocyte
0.0
0.0
KU-812 (Basophil)
57.4
67.4

none

PMA/ionomycin

2ry
0.0
0.0
CCD1106
0.0
0.0

Th1/Th2/Tr1_anti-

(Keratinocytes) none

CD95 CH11

LAK cells rest
0.0
0.0
CCD1106
0.0
0.0

(Keratinocytes)

TNF alpha + IL-1beta

LAK cells IL-2
0.0
0.0
Liver cirrhosis
100.0
100.0

LAK cells IL-2 + IL-
0.0
0.0
Lupus kidney
0.0
0.0

12

LAK cells IL-2 + IFN
0.0
0.0
NCI-H292 none
0.0
0.0

gamma

LAK cells IL-2 + IL-
0.0
0.0
NCI-H292 IL-4
0.0
9.0

18

LAK cells
0.0
5.1
NCI-H292 IL-9
0.0
0.0

PMA/ionomycin

NK Cells IL-2 rest
31.4
29.3
NCI-H292 IL-13
12.0
0.0

Two Way MLR 3
0.0
0.0
NCI-H292 IFN
2.8
0.0

day

gamma

Two Way MLR 5
0.0
0.0
HPAEC none
0.0
0.0

day

Two Way MLR 7
0.0
0.0
HPAEC TNF alpha +
31.6
27.9

day

IL-1beta

PBMC rest
0.0
0.0
Lung fibroblast none
0.0
0.0

PBMC PWM
0.0
0.0
Lung fibroblast TNF
0.0
9.9

alpha + IL-1beta

PBMC PHA-L
0.0
0.0
Lung fibroblast IL-4
0.0
15.5

Ramos (B cell) none
6.1
0.0
Lung fibroblast IL-9
6.3
23.5

Ramos (B cell)
0.0
0.0
Lung fibroblast IL-13
0.0
0.0

ionomycin

B lymphocytes PWM
0.0
0.0
Lung fibroblast IFN
0.0
0.0

gamma

B lymphocytes
0.0
7.3
Dermal fibroblast
0.0
0.0

CD40L and IL-4

CCD1070 rest

EOL-1 dbcAMP
0.0
0.0
Dermal fibroblast
0.0
0.0

CCD1070 TNF alpha

EOL-1 dbcAMP
0.0
11.0
Dermal fibroblast
0.0
0.0

PMA/ionomycin

CCD1070 IL-1beta

Dendritic cells none
0.0
0.0
Dermal fibroblast IFN
0.0
0.0

gamma

Dendritic cells LPS
16.0
9.1
Dermal fibroblast IL-4
0.0
13.3

Dendritic cells anti-
6.2
28.1
IBD Colitis 2
9.0
13.9

CD40

Monocytes rest
9.6
13.0
IBD Crohn's
7.5
4.6

Monocytes LPS
0.0
0.0
Colon
6.9
0.0

Macrophages rest
0.0
0.0
Lung
11.9
15.8

Macrophages LPS
0.0
0.0
Thymus
0.0
0.0

HUVEC none
0.0
0.0
Kidney
0.0
0.0

HUVEC starved
0.0
0.0

[0421] CNS_neurodegeneration_v1.0 Summary: Ag2354 Data from one experiment with this probe and primer set and the GMAP000818_A—1 gene is not included because of the high probability of a probe failure.

[0422] Panel 1.3D Summary: Ag2211/2221 The expression of the GMAP000818_A—1 gene is assessed in two independent runs on panel 1.3D with two different probe and primer sets. There is overall good concordance between the two runs. This gene is expressed in one melanoma cell line, two breast cancer cell lines and in placenta, testis, and prostate tissues. Thus, the expression of this gene could be used to distinguish these samples from other samples in the panel. Moreover, therapeutic modulation of this gene, through the use of small molecule drugs, antibodies or protein therapeutics might be of benefit in the treatment of melanoma or breast cancer. Ag2354 Data from one experiment with this probe and primer set and the GMAP000818_A-1 gene is not included because of the high probability of a probe failure.

[0423] Panel 2.2 Summary: Ag2211/Ag2221 The expression of the GMAP000818_A—1 gene is assessed in two independent runs on panel 2.2 using two different probe/primer sets. There is moderate concordance between the two runs. The gene is expressed highest in an ovarian cancer in one run and normal lung tissue adjacent to malignant lung tissue in the other run (CTs=31-33). In addition, there is substantial expression in bladder cancer, breast cancer and normal kidney tissue. Thus, the expression of this gene could be used to distinguish the above tissue samples from the others in the panel. Moreover, therapeutic modulation of this gene, through the use of small molecule drugs, antibodies or protein therapeutics might be of benefit in the treatment of bladder, ovarian or breast cancer.

[0424] Panel 2D Summary: Ag2354 Data from one experiment with this probe and primer set and the GMAP000818_A—1 gene is not included because of the high probability of a probe failure.

[0425] Panel 4D Summary: Ag2211/2221 Results from two experimental runs are in moderate agreement. The highest level of expression of the GMAP000818_A-1 gene in both experiments is seen in liver cirrhosis (CTs=32.5-33.5). Low level expression of this gene is also detected in IL-9 treated lung fibroblasts and HPAEC (a lung endothelial cell) treated with TNF alpha and IL-1 beta. The expression of this gene in lung derived fibroblast and endothelial cells suggests that this gene may be involved in normal conditions as well as pathological and inflammatory lung disorders that include chronic obstructive pulmonary disease, asthma, allergy and emphysema. In addition, this gene is expressed at a low but significant level in KU-812 basophil cells treated with PMA/ionomycin. These cells (KU-812) are a reasonable model for the inflammatory cells that take part in various inflammatory lung and bowel diseases, such as asthma, Crohn's disease, and ulcerative colitis. Therefore, therapeutics that modulate the function of this gene product may reduce or eliminate the symptoms of patients suffering from asthma, Crohn's disease, and ulcerative colitis. The presence of this gene in liver cirrhosis (a component of which involves liver inflammation and fibrosis) also suggests that therapeutic agents involving this gene may be useful in reducing or inhibiting the inflammation associated with fibrotic and inflammatory diseases. In addition, antibodies to the protein encoded by this gene could also be used for the diagnosis of liver cirrhosis. Ag2354 Data from one experiment with this probe and primer set and the GMAP000818_A—1 gene is not included because of the high probability of a probe failure.

[0426] D. CG54575-01/GMAC024257_A: GPCR

[0427] Expression of gene CG54575-01 was assessed using the primer-probe set Ag2209, described in Table DA. Results of the RTQ-PCR runs are shown in Tables DB, DC, DD, and DE.

14TABLE DAProbe Name Ag2209StartSEQ IDPrimersSequencesLengthPositionNO:Forward5′-acaaacctttctccctcctgta-3′22815171ProbeTET-5′-ccccatgtgcaaccccatcattt-3′-TAMRA23849172Reverse5′-atttccttgttgcggaaactat-3′22872173

[0428]

15

TABLE DB

CNS_neurodegeneration_v1.0

Rel. Exp. (%) Ag2209, Run

Rel. Exp. (%) Ag2209, Run

Tissue Name
207928566
Tissue Name
207928566

AD 1 Hippo
1.2
Control (Path) 3
3.0

Temporal Ctx

AD 2 Hippo
3.9
Control (Path) 4
6.8

Temporal Ctx

AD 3 Hippo
2.6
AD 1 Occipital Ctx
2.8

AD 4 Hippo
1.1
AD 2 Occipital Ctx
0.9

(Missing)

AD 5 Hippo
100.0
AD 3 Occipital Ctx
1.2

AD 6 Hippo
15.4
AD 4 Occipital Ctx
4.2

Control 2 Hippo
3.7
AD 5 Occipital Ctx
6.6

Control 4 Hippo
2.9
AD 5 Occipital Ctx
4.9

Control (Path) 3 Hippo
1.6
Control 1 Occipital Ctx
0.5

AD 1 Temporal Ctx
2.7
Control 2 Occipital Ctx
7.5

AD 2 Temporal Ctx
10.7
Control 3 Occipital Ctx
4.3

AD 3 Temporal Ctx
1.9
Control 4 Occipital Ctx
0.6

AD 4 Temporal Ctx
4.7
Control (Path) 1
17.0

Occipital Ctx

AD 5 Inf Temporal Ctx
10.6
Control (Path) 2
2.9

Occipital Ctx

AD 5 Sup Temporal
6.0
Control (Path) 3
0.6

Ctx

Occipital Ctx

AD 6 Inf Temporal Ctx
5.7
Control (Path) 4
4.7

Occipital Ctx

AD 6 Sup Temporal
9.7
Control 1 Parietal Ctx
4.8

Ctx

Control 1 Temporal Ctx
1.9
Control 2 Parietal Ctx
9.3

Control 2 Temporal Ctx
5.4
Control 3 Parietal Ctx
2.8

Control 3 Temporal Ctx
1.1
Control (Path) 1
10.2

Parietal Ctx

Control 3 Temporal Ctx
1.9
Control (Path) 2
3.9

Parietal Ctx

Control (Path) 1
15.2
Control (Path) 3
1.6

Temporal Ctx

Parietal Ctx

Control (Path) 2
9.3
Control (Path) 4
7.6

Temporal Ctx

Parietal Ctx

[0429]

16

TABLE DC

Panel 1.3D

Rel. Exp. (%) Ag2209, Run

Rel. Exp. (%) Ag2209, Run

Tissue Name
166003996
Tissue Name
166003996

Liver adenocarcinoma
9.5
Kidney (fetal)
15.0

Pancreas
0.0
Renal ca. 786-0
0.0

Pancreatic ca. CAPAN 2
39.8
Renal ca. A498
0.0

Adrenal gland
20.2
Renal ca. RXF 393
0.0

Thyroid
5.1
Renal ca. ACHN
3.7

Salivary gland
23.5
Renal ca. UO-31
6.7

Pituitary gland
46.3
Renal ca. TK-10
7.6

Brain (fetal)
20.2
Liver
29.9

Brain (whole)
89.5
Liver (fetal)
0.0

Brain (amygdala)
38.4
Liver ca. (hepatoblast)
0.0

HepG2

Brain (cerebellum)
100.0
Lung
0.0

Brain (hippocampus)
41.2
Lung (fetal)
12.8

Brain (substantia nigra)
24.8
Lung ca. (small cell) LX-1
51.8

Brain (thalamus)
49.7
Lung ca. (small cell)
11.7

NCI-H69

Cerebral Cortex
8.8
Lung ca. (s.cell var.)
15.7

SHP-77

Spinal cord
45.1
Lung ca. (large cell)NCI-
15.4

H460

glio/astro U87-MG
9.3
Lung ca. (non-sm. cell)
2.4

A549

glio/astro U-118-MG
0.0
Lung ca. (non-s.cell)
28.3

NCI-H23

astrocytoma SW1783
4.4
Lung ca. (non-s.cell)
18.0

HOP-62

neuro*; met SK-N-AS
10.2
Lung ca. (non-s.cl) NCI-
0.0

H522

astrocytoma SF-539
0.0
Lung ca. (squam.) SW
46.0

astrocytoma SNB-75
27.0
Lung ca. (squam.) NCI-
9.3

H596

glioma SNB-19
0.0
Mammary gland
4.0

glioma U251
0.0
Breast ca.* (pl.ef) MCF-7
0.0

glioma SF-295
9.3
Breast ca.* (pl.ef)MDA-
6.3

MB-231

Heart (Fetal)
4.4
Breast ca.* (pl. ef) T47D
20.0

Heart
0.0
Breast ca. BT-549
0.0

Skeletal muscle (Fetal)
8.8
Breast ca. MDA-N
0.0

Skeletal muscle
6.1
Ovary
0.0

Bone marrow
2.9
Ovarian ca. OVCAR-3
2.4

Thymus
27.7
Ovarian ca. OVCAR-4
0.0

Spleen
10.8
Ovarian ca. OVCAR-5
13.7

Lymph node
11.3
Ovarian ca. OVCAR-8
24.8

Colorectal
10.4
Ovarian ca. IGROV-1
23.8

Stomach
13.6
Ovarian ca. (ascites) SK-
4.0

OV-3

Small intestine
46.7
Uterus
0.0

Colon ca. SW480
0.0
Placenta
29.7

Colon ca.* SW620
36.1
Prostate
2.2

(SW480 met)

Colon ca. HT29
0.0
Prostate ca.* (bone met)
0.0

PC-3

Colon ca. HCT-116
15.7
Testis
25.2

Colon ca. CaCo-2
14.0
Melanoma Hs688(A).T
0.0

CC Well to Mod Diff
7.1
Melanoma* (met)
0.0

(ODO3866)

Hs688(B).T

Colon ca. HCC-2998
4.2
Melanoma UACC-62
0.0

Gastric ca. (liver met)
72.7
Melanoma M14
4.8

NCI-N87

Bladder
3.9
Melanoma LOX IMVI
0.0

Trachea
0.0
Melanoma* (met) SK-
4.2

MEL-5

Kidney
7.7
Adipose
12.9

[0430]

17

TABLE DD

Panel 2.2

Rel. Exp. (%) Ag2209,

Rel. Exp. (%) Ag2209,

Tissue Name
Run 174255641
Tissue Name
Run 174255641

Normal Colon
12.2
Kidney Margin (OD04348)
100.0

Colon cancer (OD06064)
5.4
Kidney malignant cancer
5.9

(OD06204B)

Colon Margin (OD06064)
3.9
Kidney normal adjacent
8.4

tissue (OD06204E)

Colon cancer (OD06159)
19.5
Kidney Cancer (OD04450-
24.3

01)

Colon Margin (OD06159)
17.9
Kidney Margin (OD04450-
41.8

03)

Colon cancer (OD06297-04)
2.8
Kidney Cancer 8120613
0.0

Colon Margin (OD06297-
15.7
Kidney Margin 8120614
2.5

015)

CC Gr.2 ascend colon
7.1
Kidney Cancer 9010320
7.3

(ODO3921)

CC Margin (ODO3921)
0.0
Kidney Margin 9010321
0.0

Colon cancer metastasis
0.0
Kidney Cancer 8120607
4.6

(OD06104)

Lung Margin (OD06104)
48.6
Kidney Margin 8120608
0.0

Colon mets to lung
0.0
Normal Uterus
17.0

(OD04451-01)

Lung Margin (OD04451-02)
24.0
Uterine Cancer 064011
19.1

Normal Prostate
8.7
Normal Thyroid
5.2

Prostate Cancer (OD04410)
0.0
Thyroid Cancer
5.1

Prostate Margin (OD04410)
0.0
Thyroid Cancer A302152
96.6

Normal Ovary
0.0
Thyroid Margin A302153
6.3

Ovarian cancer (OD06283-
10.4
Normal Breast
45.4

03)

Ovarian Margin (OD06283-
5.1
Breast Cancer
13.3

07)

Ovarian Cancer
0.0
Breast Cancer
74.7

Ovarian cancer (OD06145)
21.6
Breast Cancer (OD04590-
14.6

01)

Ovarian Margin (OD06145)
33.7
Breast Cancer Mets
3.5

(OD04590-03)

Ovarian cancer (OD06455-
13.9
Breast Cancer Metastasis
22.8

03)

Ovarian Margin (OD06455-
0.0
Breast Cancer
2.6

07)

Normal Lung
12.8
Breast Cancer 9100266
9.5

Invasive poor diff. lung
9.3
Breast Margin 9100265
22.2

adeno (ODO4945-01

Lung Margin (ODO4945-03)
39.0
Breast Cancer A209073
0.0

Lung Malignant Cancer
15.6
Breast Margin A2090734
45.1

(OD03126)

Lung Margin (OD03126)
10.9
Breast cancer (OD06083)
11.2

Lung Cancer (OD05014A)
12.4
Breast cancer node
43.2

metastasis (OD06083)

Lung Margin (OD05014B)
25.5
Normal Liver
17.1

Lung cancer (OD06081)
6.3
Liver Cancer 1026
0.0

Lung Margin (OD06081)
20.7
Liver Cancer 1025
5.0

Lung Cancer (OD04237-01)
0.0
Liver Cancer 6004-T
18.2

Lung Margin (OD04237-02)
4.4
Liver Tissue 6004-N
0.0

Ocular Mel Met to Liver
68.8
Liver Cancer 6005-T
0.0

(ODO4310)

Liver Margin (ODO4310)
0.0
Liver Tissue 6005-N
0.0

Melanoma Metastasis
0.0
Liver Cancer
10.5

Lung Margin (OD04321)
15.6
Normal Bladder
9.3

Normal Kidney
20.4
Bladder Cancer
0.0

Kidney Ca, Nuclear grade 2
41.8
Bladder Cancer
0.0

(OD04338)

Kidney Margin (OD04338)
10.8
Normal Stomach
20.7

Kidney Ca Nuclear grade 1/2
25.9
Gastric Cancer 9060397
0.0

(OD04339)

Kidney Margin (OD04339)
39.2
Stomach Margin 9060396
13.8

Kidney Ca, Clear cell type
8.7
Gastric Cancer 9060395
4.4

(OD04340)

Kidney Margin (OD04340)
15.5
Stomach Margin 9060394
14.3

Kidney Ca, Nuclear grade 3
0.0
Gastric Cancer 064005
11.7

(OD04348)

[0431]

18

TABLE DE

Panel 4D

Rel. Exp. (%) Ag2209,

Rel. Exp. (%) Ag2209,

Tissue Name
Run 164331296
Tissue Name
Run 164331296

Secondary Th1 act
8.9
HUVEC IL-1beta
0.0

Secondary Th2 act
10.2
HUVEC IFN gamma
2.6

Secondary Tr1 act
8.2
HUVEC TNF alpha + IFN
0.0

gamma

Secondary Th1 rest
17.3
HUVEC TNF alpha + IL4
1.8

Secondary Th2 rest
15.6
HUVEC IL-11
2.7

Secondary Tr1 rest
18.2
Lung Microvascular EC none
1.3

Primary Th1 act
17.9
Lung Microvascular EC
1.1

TNFalpha + IL-1beta

Primary Th2 act
25.5
Microvascular Dermal EC none
1.8

Primary Tr1 act
25.9
Microsvasular Dermal EC
0.0

TNFalpha + IL-1beta

Primary Th1 rest
100.0
Bronchial epithelium
13.6

TNFalpha + IL1beta

Primary Th2 rest
54.0
Small airway epithelium none
4.8

Primary Tr1 rest
29.3
Small airway epithelium
29.1

TNFalpha + IL-1beta

CD45RA CD4 lymphocyte
6.5
Coronery artery SMC rest
4.2

CD45RO CD4 lymphocyte
29.5
Coronery artery SMC
2.1

act

TNFalpha + IL-1beta

CD8 lymphocyte act
15.6
Astrocytes rest
2.7

Secondary CD8
31.0
Astrocytes TNFalpha + IL-1beta
1.0

lymphocyte rest

Secondary CD8
18.6
KU-812 (Basophil) rest
6.2

lymphocyte act

CD4 lymphocyte none
9.8
(KU-812 (Basophil)
15.5

PMA/ionomycin

2ry Th1/Th2/Tr1_anti-
38.4
CCD1106 (Keratinocytes) none
8.7

CD95 CH11

LAK cells rest
21.0
CCD1106 (Keratinocytes)
4.5

TNFalpha + IL-1beta

LAK cells IL-2
58.2
Liver cirrhosis
5.8

LAK cells IL-2 + IL-12
48.6
Lupus kidney
6.7

LAK cells IL-2 + IFN
85.9
NCI-H292 none
44.1

gamma

LAK cells IL-2 + IL-18
37.9
NCI-H292 IL-4
40.6

LAK cells
1.0
NCI-H292 IL-9
35.6

PMA/ionomycin

NK Cells IL-2 rest
29.5
NCI-H292 IL-13
9.9

Two Way MLR 3 day
57.8
NCI-H292 IFN gamma
22.1

Two Way MLR 5 day
18.7
HPAEC none
0.9

Two Way MLR 7 day
17.3
HPAEC TNF alpha + IL-1 beta
0.0

PBMC rest
2.5
Lung fibroblast none
1.4

PBMC PWM
84.7
Lung fibroblast TNF alpha + IL-
1.0

1 beta

PBMC PHA-L
30.6
Lung fibroblast IL-4
3.3

Ramos (B cell) none
2.9
Lung fibroblast IL-9
5.8

Ramos (B cell) ionomycin
6.3
Lung fibroblast IL-13
0.0

B lymphocytes PWM
76.3
Lung fibroblast IFN gamma
1.5

B lymphocytes CD40L
49.7
Dermal fibroblast CCD1070 rest
10.1

and IL-4

EOL-1 dbcAMP
8.5
Dermal fibroblast CCD1070
43.2

TNF alpha

EOL-1 dbcAMP
5.4
Dermal fibroblast CCD1070 IL-
6.7

PMA/ionomycin

1 beta

Dendritic cells none
16.6
Dermal fibroblast IFN gamma
1.2

Dendritic cells LPS
25.0
Dermal fibroblast IL-4
7.2

Dendritic cells anti-CD40
24.5
IBD Colitis 2
4.2

Monocytes rest
6.7
IBD Crohn's
1.7

Monocytes LPS
6.1
Colon
14.3

Macrophages rest
29.5
Lung
3.9

Macrophages LPS
8.4
Thymus
21.0

HUVEC none
1.0
Kidney
45.7

HUVEC starved
2.9

[0432] CNS_neurodegeneration_v1.0 Summary: Ag2209 No difference was detected in the expression of the CG54575-01 gene in the postmortem brains of Alzheimer's diseased patients when compared to controls; however this panel demonstrates the expression of this gene in the brains of an independent group of subjects. Please see panel 1.3d for a discussion of utility of this gene in the central nervous system.

[0433] Panel 1.3D Summary: Ag2209 Expression of the GPCR encoded by the CG54575-01 gene is highest in the brain, especially in the cerebellum and (to a lesser extent) in the hippocampus. The mRNA was not detected in Panel CNSD.01 as neither of these regions is represented on that panel. Cerebellum-specific targets are of great interest in the treatment of spinocerebellar ataxias, a group of glutamate-repeat disorders which cause neurodegeneration in the cerebellum and are often fatal. Selective stimulation of cerebellar neurons may result in the lessening of the major symptoms of these disorders (ataxia) and could possibly slow neurodegeneration. Furthermore, because this GPCR is found in the hippocampus (a region of the brain critical for long-term memory formation), selective stimulation may enhance memory in various forms of senile dementia (Alzheimer's type, non-Alzheimer's type, vascular dementia, and others). Expression of the CG54575-01 gene is also detectable to a much lesser extent in thalamus, spinal cord, pituitary gland, and small intestine.

[0434] Panel 2.2 Summary: Ag2209 The expression of the CG54575-01 gene appears to be widespread at low levels across the samples on this panel. Interestingly, there seems to be an association of expression of this gene with a number of normal tissues when compared with their cancerous counterparts. This specifically seems to be the case in gastric, breast, kidney, lung and colon. Thus, therapeutic modulation of this gene might be of utility in the treatment of the above listed cancers.

[0435] Panel 4D Summary: Ag2209 The CG54575-01 transcript is highly expressed in activated B cells, LAK cells and primary resting Th1 and Th2 T cells. High expression of this transcript is restricted to activated B cells (B cells treated with PWM or CD40L+IL4); it is not expressed in the B cell line Ramos. The expression of this transcript in PBMC treated with the B cell mitogen, PWM, confirms the importance of CG54575-01 gene expression in activated B cells. In addition, this transcript is also abundantly expressed on primary resting Th1 cells (to a lesser degree on primary resting Th2 cells). Therefore, it appears that the CG54575-01 gene, encoding a GPCR homolog, is a potential new member of the chemokine receptor family. The expression of this protein in activated B cells suggests a role for this protein in their trafficking to appropriate sites where they can fully activate antigen specific T cells. Thus, the CG54575-01 protein is likely to participate in the development of immune or inflammatory reactions. In addition, this transcript is highly expressed on activated LAK cells, predominantly on LAK cells treated with I1-2 and IFN-g, suggesting a role for this protein in tumor surveillance. Therapeutic modulation of the CG54575-01 gene with monoclonal antibodies or small molecule therapeutics could be valuable in the treatment of immunological disorders, immune rejection and tumors. Moderate expression of the CG54575-01 transcript in H292 cells and activated dermal fibroblast (TNFa), suggests that therapeutic modulation of this gene might also be useful in asthma and emphysema.

[0436] Panel CNS—1 Summary: Ag2209 Expression of the CG54575-01 gene is low to undetectable across the samples in this panel. However, this is probably due to the absence of cerebellum and hippocampus samples on this panel. (Data not shown.) Please see Panel 1.3D for discussion of the potential utility of this gene in neurological function.

[0437] E. CG56103-03/GMAC011537_A: Olfactory Receptor

[0438] Expression of gene was assessed using the primer-probe set Ag2205, described in Table EA. Results of the RTQ-PCR runs are shown in Tables EB and EC.

19TABLE EAProbe Name Ag2205StartSEQ IDPrimersSequencesLengthPositionNO:Forward5′-acctccgtgtctgaattcatc-3′2130174ProbeTET-5′-ccacctccagctgatgctcttcct-3′-TAMRA2474175Reverse5′-acaggtacatcagcaggaacag-3′2299176

[0439]

20

TABLE EB

Panel 1.3D

Rel. Exp. (%) Ag2205, Run

Rel. Exp. (%) Ag2205, Run

Tissue Name
165974832
Tissue Name
165974832

Liver adenocarcinoma
0.0
Kidney (fetal)
0.0

Pancreas
0.0
Renal ca. 786-0
0.0

Pancreatic ca. CAPAN 2
0.0
Renal ca. A498
0.0

Adrenal gland
0.0
Renal ca. RXF 393
0.0

Thyroid
0.0
Renal ca. ACHN
0.0

Salivary gland
0.0
Renal ca. UO-31
100.0

Pituitary gland
0.0
Renal ca. TK-10
0.0

Brain (fetal)
0.0
Liver
0.0

Brain (whole)
0.0
Liver (fetal)
0.0

Brain (amygdala)
0.0
Liver ca. (hepatoblast)
0.0

HepG2

Brain (cerebellum)
19.6
Lung
0.0

Brain (hippocampus)
25.3
Lung (fetal)
0.0

Brain (substantia nigra)
0.0
Lung ca. (small cell) LX-1
0.0

Brain (thalamus)
15.5
Lung ca. (small cell)
0.0

NCI-H69

Cerebral Cortex
0.0
Lung ca. (s.cell var.)
0.0

SHP-77

Spinal cord
18.3
Lung ca. (large cell)NCI-
0.0

H460

glio/astro U87-MG
0.0
Lung ca. (non-sm. cell)
0.0

A549

glio/astro U-118-MG
0.0
Lung ca. (non-s.cell)
0.0

NCI-H23

astrocytoma SW1783
0.0
Lung ca. (non-s.cell)
46.3

HOP-62

neuro*; met SK-N-AS
22.7
Lung ca. (non-s.cl) NCI-
0.0

H522

astrocytoma SF-539
0.0
Lung ca. (squam.) SW
0.0

900

astrocytoma SNB-75
0.0
Lung ca. (squam.) NCI-
0.0

H596

glioma SNB-19
0.0
Mammary gland
0.0

glioma U251
0.0
Breast ca.* (pl.ef) MCF-7
0.0

glioma SF-295
16.2
Breast ca.* (pl.ef) MDA-
0.0

MB-231

Heart (Fetal)
0.0
Breast ca.* (pl. ef) T47D
0.0

Heart
0.0
Breast ca. BT-549
0.0

Skeletal muscle (Fetal)
0.0
Breast ca. MDA-N
0.0

Skeletal muscle
0.0
Ovary
0.0

Bone marrow
0.0
Ovarian ca. OVCAR-3
17.7

Thymus
0.0
Ovarian ca. OVCAR-4
2.0

Spleen
0.0
Ovarian ca. OVCAR-5
0.0

Lymph node
0.0
Ovarian ca. OVCAR-8
0.0

Colorectal
5.4
Ovarian ca. IGROV-1
0.0

Stomach
0.0
Ovarian ca. (ascites) SK-
14.7

OV-3

Small intestine
0.0
Uterus
0.0

Colon ca. SW480
0.0
Placenta
0.0

Colon ca.* SW620
0.0
Prostate
0.0

(SW480 met)

Colon ca. HT29
0.0
Prostate ca.* (bone met)
0.0

PC-3

Colon ca. HCT-116
0.0
Testis
1.3

Colon ca. CaCo-2
0.0
Melanoma Hs688(A).T
0.0

CC Well to Mod Diff
0.0
Melanoma* (met)
0.0

(ODO3866)

Hs688(B).T

Colon ca. HCC-2998
0.0
Melanoma UACC-62
0.0

Gastric ca. (liver met)
24.0
Melanoma M14
0.0

NCI-N87

Bladder
0.0
Melanoma LOX IMVI
0.0

Trachea
0.0
Melanoma* (met) SK-
0.0

MEL-5

Kidney
0.0
Adipose
0.0

[0440]

21

TABLE EC

Panel 4D

Rel. Exp. (%) Ag2205,

Rel. Exp. (%) Ag2205,

Tissue Name
Run 163623519
Tissue Name
Run 163623519

Secondary Th1 act
0.0
HUVEC IL-1beta
0.0

Secondary Th2 act
0.0
HUVEC IFN gamma
0.0

Secondary Tr1 act
0.0
HUVEC TNF alpha + IFN
0.0

gamma

Secondary Th1 rest
0.0
HUVEC TNF alpha + IL4
0.0

Secondary Th2 rest
0.0
HUVEC IL-11
8.4

Secondary Tr1 rest
0.0
Lung Microvascular EC none
0.0

Primary Th1 act
0.0
Lung Microvascular EC
0.0

TNFalpha + IL-1beta

Primary Th2 act
0.0
Microvascular Dermal EC none
0.0

Primary Tr1 act
0.0
Microsvasular Dermal EC
0.0

TNFalpha + IL-1beta

Primary Th1 rest
0.0
Bronchial epithelium
0.0

TNFalpha + IL1beta

Primary Th2 rest
0.0
Small airway epithelium none
0.0

Primary Tr1 rest
5.5
Small airway epithelium
0.0

TNFalpha + IL-1beta

CD45RA CD4 lymphocyte
0.0
Coronery artery SMC rest
0.0

act

CD45RO CD4 lymphocyte
15.5
Coronery artery SMC
0.0

act

TNFalpha + IL-1beta

CD8 lymphocyte act
7.0
Astrocytes rest
0.0

Secondary CD8
0.0
Astrocytes TNFalpha + IL-1beta
0.0

lymphocyte rest

Secondary CD8
0.0
KU-812 (Basophil) rest
0.0

lymphocyte act

CD4 lymphocyte none
0.0
KU-812 (Basophil)
0.0

PMA/ionomycin

2ry Th1/Th2/Tr1_anti-
0.0
CCD1106 (Keratinocytes) none
0.0

CD95 CH11

LAK cells rest
0.0
CCD1106 (Keratinocytes)
0.0

TNFalpha + IL-1beta

LAK cells IL-2
0.0
Liver cirrhosis
100.0

LAK cells IL-2 + IL-12
15.1
Lupus kidney
0.0

LAK cells IL-2 + IFN
0.0
NCI-H292 none
0.0

gamma

LAK cells IL-2 + IL-18
5.9
NCI-H292 IL-4
0.0

LAK cells
0.0
NCI-H292 IL-9
0.0

PMA/ionomycin

NK Cells IL-2 rest
0.0
NCI-H292 IL-13
0.0

Two Way MLR 3 day
0.0
NCI-H292 IFN gamma
0.0

Two Way MLR 5 day
9.6
HPAEC none
0.0

Two Way MLR 7 day
14.6
HPAEC TNF alpha + IL-1 beta
0.0

PBMC rest
0.0
Lung fibroblast none
0.0

PBMC PWM
0.0
Lung fibroblast TNF alpha + IL-
0.0

1 beta

PBMC PHA-L
0.0
Lung fibroblast IL-4
0.0

Ramos (B cell) none
0.0
Lung fibroblast IL-9
0.0

Ramos (B cell) ionomycin
0.0
Lung fibroblast IL-13
0.0

B lymphocytes PWM
8.2
Lung fibroblast IFN gamma
0.0

B lymphocytes CD40L
0.0
Dermal fibroblast CCD1070 rest
0.0

and IL-4

EOL-1 dbcAMP
0.0
Dermal fibroblast CCD1070
0.0

TNF alpha

EOL-1 dbcAMP
0.0
Dermal fibroblast CCD1070 IL-
0.0

PMA/ionomycin

1 beta

Dendritic cells none
0.0
Dermal fibroblast IFN gamma
0.0

Dendritic cells LPS
0.0
Dermal fibroblast IL-4
0.0

Dendritic cells anti-CD40
0.0
IBD Colitis 2
8.2

Monocytes rest
0.0
IBD Crohn's
6.9

Monocytes LPS
0.0
Colon
0.0

Macrophages rest
0.0
Lung
54.0

Macrophages LPS
0.0
Thymus
0.0

HUVEC none
0.0
Kidney
0.0

HUVEC starved
0.0

[0441] CNS_neurodegeneration_v1.0 Summary: Ag2205 Expression of the CG56103-02 gene is low/undetectable across all of the samples on this panel. (Data not shown.)

[0442] Panel 1.3D Summary: Ag2205 Significant expression of the CG56103-02 gene is limited to a renal cancer cell line and a lung cancer cell line (CTs=33-34). Thus, expression of this gene could be used to differentiate between these cell lines and other samples on this panel and as a marker to detect the presence of lung and renal cancer.

[0443] Panel 2.2 Summary: Ag2205 Expression of the CG56103-02 gene is low/undetectable across all of the samples on this panel. (Data not shown.)

[0444] Panel 4D Summary: Ag2205 Low but significant expression of the CG56103-02 gene is detected in a liver cirrhosis sample (CT=33.46). Furthermore, expression of this gene is not detected in normal liver in Panel 1.3D, suggesting that its expression is unique to liver cirrhosis. This gene encodes a putative GPCR; therefore, antibodies or small molecule therapeutics could reduce or inhibit fibrosis that occurs in liver cirrhosis. In addition, antibodies to this putative GPCR could also be used for the diagnosis of liver cirrhosis. In addition, expression in normal lung suggests a possible role in lung homeostasis.

[0445] F. CG54362-02/GMAP001804_E: Olfactory Receptor

[0446] Expression of gene CG54362-01 was assessed using the primer-probe sets Ag2358, Ag2359 and Ag1640, described in Tables FA, FB and FC. Results of the RTQ-PCR runs are shown in Table FD.

22TABLE FAProbe Name Ag2358StartSEQ IDPrimersSequencesLengthPositionNO:Forward5′-ccatgtcagtgagctggtattt-3′22574177ProbeTET-5′-tggagtaatcaccatgctatccagca-3′-TAMRA26607178Reverse5′-tcaaagcgtaagagatgacgat-3′22638179

[0447]

23

TABLE FB

Probe Name Ag2359

Start
SEQ ID

Primers
Sequences
Length
Position
NO:

Forward
5′-ccatgtcagtgagctggtattt-3′
22
574
180

Probe
TET-5′-tggagtaatcaccatgctatccagca-3′-TAMRA
26
607
181

Reverse
5′-tcaaagcgtaagagatgacgat-3′
22
638
182

[0448]

24

TABLE FC

Probe Name Ag1640

Start
SEQ ID

Primers
Sequences
Length
Position
NO:

Forward
5′-ccatgtcagtgagctggtattt-3′
22
574
183

Probe
TET-5′-tggagtaatcaccatgctatccagca-3′-TAMRA
26
607
184

Reverse
5′-tcaaagcgtaagagatgacgat-3′
22
638
185

[0449]

25

TABLE FD

Panel 2D

Rel. Exp. (%)
Rel. Exp. (%)

Rel. Exp. (%)
Rel. Exp. (%)

Ag2358, Run
Ag2359, Run

Ag2358, Run
Ag2359, Run

Tissue Name
164025336
164151618
Tissue Name
164025336
164151618

Normal Colon
8.0
2.3
Kidney Margin
0.0
0.0

8120608

CC Well to Mod
4.4
4.2
Kidney Cancer
0.0
0.0

Diff (ODO3866)

8120613

CC Margin
1.7
9.2
Kidney Margin
0.0
0.0

(ODO3866)

8120614

CC Gr.2
0.0
0.0
Kidney Cancer
0.0
0.0

rectosigmoid

9010320

(ODO3868)

CC Margin
0.0
0.0
Kidney Margin
0.0
0.0

(ODO3868)

9010321

CC Mod Diff
0.0
0.0
Normal Uterus
0.8
0.0

(ODO3920)

CC Margin
0.0
0.0
Uterine Cancer
0.0
0.0

(ODO3920)

064011

CC Gr.2 ascend
0.0
0.0
Normal Thyroid
0.0
0.0

colon (ODO3921)

CC Margin
1.1
1.2
Thyroid Cancer
0.0
0.0

(ODO3921)

CC from Partial
0.0
0.0
Thyroid Cancer
0.0
0.0

Hepatectomy

A302152

(ODO4309) Mets

Liver Margin
0.0
0.0
Thyroid Margin
0.0
0.0

(ODO4309)

A302153

Colon mets to lung
4.5
0.0
Normal Breast
0.0
0.0

(OD04451-01)

Lung Margin
0.0
0.0
Breast Cancer
0.0
0.0

(OD04451-02)

Normal Prostate
0.0
0.0
Breast Cancer
0.0
0.0

6546-1

(OD04590-01)

Prostate Cancer
0.0
0.0
Breast Cancer
0.0
2.2

(OD04410)

Mets (OD04590-

03)

Prostate Margin
0.0
0.0
Breast Cancer
0.0
0.0

(OD04410)

Metastasis

Prostate Cancer
0.0
0.0
Breast Cancer
0.0
1.7

(OD04720-01)

Prostate Margin
0.0
0.0
Breast Cancer
3.3
0.0

(OD04720-02)

Normal Lung
0.0
0.0
Breast Cancer
0.0
0.0

9100266

Lung Met to Muscle
0.0
2.1
Breast Margin
0.0
0.0

(ODO4286)

9100265

Muscle Margin
0.0
0.0
Breast Cancer
1.8
0.0

(ODO4286)

A209073

Lung Malignant
1.3
0.0
Breast Margin
0.0
0.0

Cancer (OD03126)

A2090734

Lung Margin
0.9
0.0
Normal Liver
0.0
0.0

(OD03126)

Lung Cancer
0.0
0.0
Liver Cancer
6.5
21.6

(OD04404)

Lung Margin
0.0
0.0
Liver Cancer
0.0
0.0

(OD04404)

1025

Lung Cancer
0.0
0.0
Liver Cancer
0.0
0.0

(OD04565)

1026

Lung Margin
0.0
0.0
Liver Cancer
0.0
1.4

(OD04565)

6004-T

Lung Cancer
1.7
0.0
Liver Tissue
0.0
0.0

(OD04237-01)

6004-N

Lung Margin
0.0
0.0
Liver Cancer
0.0
0.0

(OD04237-02)

6005-T

Ocular Mel Met to
0.0
1.8
Liver Tissue
0.0
0.0

Liver (ODO4310)

6005-N

Liver Margin
0.0
0.0
Normal Bladder
0.0
0.0

(ODO4310)

Melanoma
0.0
0.0
Bladder Cancer
0.0
0.0

Metastasis

Lung Margin
0.0
0.0
Bladder Cancer
57.8
45.7

(OD04321)

Normal Kidney
5.1
3.4
Bladder Cancer
0.0
0.0

(OD04718-01)

Kidney Ca, Nuclear
13.4
8.1
Bladder Normal
0.0
0.0

grade 2 (OD04338)

Adjacent

(OD04718-03)

Kidney Margin
4.2
0.0
Normal Ovary
0.0
0 0

(OD04338)

Kidney Ca Nuclear
5.6
0.0
Ovarian Cancer
0.0
0.0

grade 1/2

(OD04339)

Kidney Margin
0.0
1.6
Ovarian Cancer
100.0
100.0

(OD04339)

(OD04768-07)

Kidney Ca, Clear
0.0
0.0
Ovary Margin
0.0
0.0

cell type (OD04340)

(OD04768-08)

Kidney Margin
0.0
3.7
Normal Stomach
0.0
3.5

(OD04340)

Kidney Ca, Nuclear
0.0
0.0
Gastric Cancer
0.0
0.0

grade 3 (OD04348)

9060358

Kidney Margin
0.0
0.0
Stomach Margin
0.0
0.0

(OD04348)

9060359

Kidney Cancer
0.0
0.0
Gastric Cancer
0.0
0.0

(OD04622-01)

9060395

Kidney Margin
0.0
0.0
Stomach Margin
0.0
0.0

(OD04622-03)

9060394

Kidney Cancer
0.0
0.0
Gastric Cancer
0.0
0.0

(OD04450-01)

9060397

Kidney Margin
0.0
0.0
Stomach Margin
0.0
0.0

(OD04450-03)

9060396

Kidney Cancer
0.0
0.0
Gastric Cancer
1.8
0.0

8120607

064005

[0450] CNS_neurodegeneration_v1.0 Summary: Ag2358/Ag2359 Expression of the CG54362-02 gene is low/undetectable (CTs>35) in all samples on this panel. (Data not shown.)

[0451] Panel 1.3D Summary: Ag1640/Ag2358/Ag2359 Expression of the CG54362-02 gene is low/undetectable (CTs>35) in all samples on this panel. (Data not shown.)

[0452] Panel 2D Summary: Ag2359/Ag2358 Two experiments with the same probe and primer set produce results that are in excellent agreement, with highest expression of the CG54362-02 gene in a sample of ovarian cancer with limited (CTs=-32-33). Thus, this gene may be useful in distinguishing ovarian cancers from other tissues. Therapeutic modulation of this gene may also be useful in the treatment of ovarian cancers.

[0453] Panel 4D Summary: Ag1640/Ag2359/Ag2358 Expression of the CG54362-02 gene is low/undetectable (CT values>35) in all samples on this panel.

[0454] G. CG54335-01/GMAP001804_A: GPCR

[0455] Expression of gene CG54335-01 was assessed using the primer-probe sets Ag2355 and Ag1635, described in Tables GA and GB. Results of the RTQ-PCR runs are shown in Tables GC, GD, GE, and GF.

26TABLE GAProbe Name Ag2355StartSEQ IDPrimersSequencesLengthPositionNO:Forward5′-tcatacaagtgccatgatgaaa-3′22478186ProbeTET-5′-tgtccttttgcaaatcccacattatca-3′-TAMRA27501187Reverse5′-aggggaagaacatcacagaagt-3′22534188

[0456]

27

TABLE GB

Probe Name Ag1635

Start
SEQ ID

Primers
Sequences
Length
Position
NO:

Forward
5′-tcatacaagtgccatgatgaaa-3′
22
478
189

Probe
TET-5′-tgtccttttgcaaatcccacattatca-3′-TAMRA
27
501
190

Reverse
5′-aggggaagaacatcacagaagt-3′
22
534
191

[0457]

28

TABLE GC

Panel 1.3D

Rel. Exp. (%) Ag2355, Run

Rel. Exp. (%) Ag2355, Run

Tissue Name
166005271
Tissue Name
166005271

Liver adenocarcinoma
0.0
Kidney (fetal)
0.0

Pancreas
0.0
Renal ca. 786-0
0.0

Pancreatic ca. CAPAN 2
0.0
Renal ca. A498
0.0

Adrenal gland
0.0
Renal ca. RXF 393
0.0

Thyroid
0.0
Renal ca. ACHN
0.0

Salivary gland
0.0
Renal ca. UO-31
0.0

Pituitary gland
0.0
Renal ca. TK-10
0.0

Brain (fetal)
0.0
Liver
0.0

Brain (whole)
0.0
Liver (fetal)
0.0

Brain (amygdala)
0.0
Liver ca. (hepatoblast)
0.0

HepG2

Brain (cerebellum)
0.0
Lung
0.0

Brain (hippocampus)
0.0
Lung (fetal)
0.0

Brain (substantia nigra)
0.0
Lung ca. (small cell) LX-1
0.0

Brain (thalamus)
0.0
Lung ca. (small cell)
0.0

NCI-H69

Cerebral Cortex
0.0
Lung ca. (s.cell var.)
0.0

SHP-77

Spinal cord
0.0
Lung ca. (large cell)NCI-
0.0

H460

glio/astro U87-MG
8.1
Lung ca. (non-sm. cell)
0.0

A549

glio/astro U-118-MG
0.0
Lung ca. (non-s.cell)
0.0

NCI-H23

astrocytoma SW1783
0.0
Lung ca. (non-s.cell)
7.1

HOP-62

neuro*; met SK-N-AS
0.0
Lung ca. (non-s.cl) NCI-
0.0

H522

astrocytoma SF-539
0.0
Lung ca. (squam.) SW
7.7

900

astrocytoma SNB-75
0.0
Lung ca. (squam.) NCI-
0.0

H596

glioma SNB-19
6.4
Mammary gland
0.0

glioma U251
0.0
Breast ca.* (pl.ef) MCF-7
100.0

glioma SF-295
0.0
Breast ca.* (pl.ef) MDA-
0.0

MB-231

Heart (Fetal)
0.0
Breast ca.* (pl. ef) T47D
57.4

Heart
0.0
Breast ca. BT-549
0.0

Skeletal muscle (Fetal)
0.0
Breast ca. MDA-N
0.0

Skeletal muscle
6.1
Ovary
0.0

Bone marrow
0.0
Ovarian ca. OVCAR-3
0.0

Thymus
0.0
Ovarian ca. OVCAR-4
0.0

Spleen
0.0
Ovarian ca. OVCAR-5
0.0

Lymph node
0.0
Ovarian ca. OVCAR-8
0.0

Colorectal
0.0
Ovarian ca. IGROV-1
0.0

Stomach
0.0
Ovarian ca. (ascites) SK-
0.0

OV-3

Small intestine
0.0
Uterus
0.0

Colon ca. SW480
0.0
Placenta
19.2

Colon ca.* SW620
0.0
Prostate
0.0

(SW480 met)

Colon ca. HT29
0.0
Prostate ca.* (bone met)
4.7

PC-3

Colon ca. HCT-116
0.0
Testis
53.2

Colon ca. CaCo-2
0.0
Melanoma Hs688(A).T
0.0

CC Well to Mod Diff
0.0
Melanoma* (met)
0.0

(ODO3866)

Hs688(B).T

Colon ca. HCC-2998
0.0
Melanoma UACC-62
11.0

Gastric ca. (liver met)
0.0
Melanoma M14
0.0

NCI-N87

Bladder
0.0
Melanoma LOX IMVI
0.0

Trachea
0.0
Melanoma* (met) SK-
0.0

MEL-5

Kidney
0.0
Adipose
0.0

[0458]

29

TABLE GD

Panel 2D

Rel. Exp. (%) Ag2355,

Rel. Exp. (%) Ag2355,

Tisue Name
Run 164025176
Tissue Name
Run 164025176

Normal Colon
4.4
Kidney Margin 8120608
0.0

CC Well to Mod Diff
0.7
Kidney Cancer 8120613
0.0

(ODO3866)

CC Margin (ODO3866)
3.7
Kidney Margin 8120614
0.4

CC Gr.2 rectosigmoid
0.0
Kidney Cancer 9010320
0.4

(ODO3868)

CC Margin (ODO3868)
0.0
Kidney Margin 9010321
0.0

CC Mod Diff (ODO3920)
0.0
Normal Uterus
1.1

CC Margin (ODO3920)
0.3
Uterine Cancer 064011
0.0

CC Gr.2 ascend colon
0.3
Normal Thyroid
0.0

(ODO3921)

CC Margin (ODO3921)
1.4
Thyroid Cancer
0.0

CC from Partial Hepatectomy
0.0
Thyroid Cancer A302152
0.0

(ODO4309) Mets

Liver Margin (ODO4309)
0.0
Thyroid Margin A302153
0.0

Colon mets to lung (OD04451-
0.0
Normal Breast
0.0

01)

Lung Margin (OD04451-02)
0.8
Breast Cancer
0.0

Normal Prostate 6546-1
0.8
Breast Cancer (OD04590-
0.0

01)

Prostate Cancer (OD04410)
0.0
Breast Cancer Mets
0.0

(OD04590-03)

Prostate Margin (OD04410)
0.0
Breast Cancer Metastasis
0.0

Prostate Cancer (OD04720-01)
0.0
Breast Cancer
0.0

Prostate Margin (OD04720-02)
0.5
Breast Cancer
0.0

Normal Lung
1.8
Breast Cancer 9100266
0.0

Lung Met to Muscle
0.8
Breast Margin 9100265
0.4

(ODO4286)

Muscle Margin (ODO4286)
0.0
Breast Cancer A209073
0.0

Lung Malignant Cancer
0.0
Breast Margin A2090734
0.4

(OD03126)

Lung Margin (OD03126)
0.0
Normal Liver
0.0

Lung Cancer (OD04404)
0.0
Liver Cancer
5.0

Lung Margin (OD04404)
0.0
Liver Cancer 1025
0.7

Lung Cancer (OD04565)
0.0
Liver Cancer 1026
0.0

Lung Margin (OD04565)
0.0
Liver Cancer 6004-T
0.3

Lung Cancer (OD04237-01)
0.3
Liver Tissue 6004-N
0.7

Lung Margin (OD04237-02)
0.0
Liver Cancer 6005-T
0.0

Ocular Mel Met to Liver
0.0
Liver Tissue 6005-N
0.0

(ODO4310)

Liver Margin (ODO4310)
0.0
Normal Bladder
1.8

Melanoma Metastasis
0.0
Bladder Cancer
0.0

Lung Margin (OD04321)
0.0
Bladder Cancer
18.2

Normal Kidney
0.9
Bladder Cancer
0.0

(OD04718-01)

Kidney Ca, Nuclear grade 2
2.1
Bladder Normal Adjacent
0.0

(OD04338)

(OD04718-03)

Kidney Margin (OD04338)
0.0
Normal Ovary
0.0

Kidney Ca Nuclear grade 1/2
0.8
Ovarian Cancer
0.0

(OD04339)

Kidney Margin (OD04339)
0.8
Ovarian Cancer
100.0

(OD04768-07)

Kidney Ca, Clear cell type
0.0
Ovary Margin (OD04768-
0.0

(OD04340)

08)

Kidney Margin (OD04340)
0.0
Normal Stomach
0.2

Kidney Ca, Nuclear grade 3
0.0
Gastric Cancer 9060358
0.0

(OD04348)

Kidney Margin (OD04348)
0.0
Stomach Margin 9060359
0.0

Kidney Cancer (OD04622-01)
0.0
Gastric Cancer 9060395
0.0

Kidney Margin (OD04622-03)
0.0
Stomach Margin 9060394
0.0

Kidney Cancer (OD04450-01)
0.0
Gastric Cancer 9060397
0.0

Kidney Margin (OD04450-03)
0.0
Stomach Margin 9060396
0.0

Kidney Cancer 8120607
0.0
Gastric Cancer 064005
0.7

[0459]

30

TABLE GE

Panel 3D

Rel. Exp. (%)

Rel. Exp. (%)

Ag2355, Run

Ag2355, Run

Tissue Name
164843783
Tissue Name
164843783

Daoy-Medulloblastoma
0.0
Ca Ski-Cervical epidermoid
0.0

carcinoma (metastasis)

TE671-Medulloblastoma
13.5
ES-2-Ovarian clear cell carcinoma
0.0

D283 Med-Medulloblastoma
0.0
Ramos-Stimulated with
0.0

PMA/ionomycin 6h

PFSK-1-Primitive
0.0
Ramos-Stimulated with
0.0

Neuroectodermal

PMA/ionomycin 14h

XF-498-CNS
0.0
MEG-01-Chronic myelogenous
0.0

leukemia (megokaryoblast)

SNB-78-Glioma
0.0
Raji-Burkitt's lymphoma
0.0

SF-268-Glioblastoma
0.0
Daudi-Burkitt's lymphoma
0.0

T98G-Glioblastoma
0.0
U266-B-cell plasmacytoma
72.7

SK-N-SH-Neuroblastoma
0.0
CA46-Burkitt's lymphoma
0.0

(metastasis)

SF-295-Glioblastoma
0.0
RL-non-Hodgkin's B-cell
0.0

lymphoma

Cerebellum
0.0
JM1-pre-B-cell lymphoma
0.0

Cerebellum
0.0
Jurkat-T cell leukemia
0.0

NCI-H292-Mucoepidermoid
0.0
TF-1-Erythroleukemia
0.0

lung carcinoma

DMS-114-Small cell lung
57.8
HUT 78-T-cell lymphoma
3.6

cancer

DMS-79-Small cell lung
0.0
U937-Histiocytic lymphoma
0.0

cancer

NCI-H146-Small cell lung
4.9
KU-812-Myelogenous leukemia
0.0

cancer

NCI-H526-Small cell lung
4.1
769-P-Clear cell renal carcinoma
0.0

cancer

NCI-N417-Small cell lung
0.0
Caki-2-Clear cell renal carcinoma
0.0

cancer

NCI-H82-Small cell lung
0.0
SW 839-Clear cell renal carcinoma
0.0

cancer

NCI-H157-Squamous cell
28.9
G401-Wilms' tumor
0.0

lung cancer (metastasis)

NCI-H1155-Large cell lung
0.0
Hs766T-Pancreatic carcinoma (LN
0.0

cancer

metastasis)

NCI-H1299-Large cell lung
100.0
CAPAN-1-Pancreatic
0.0

cancer

adenocarcinoma (liver metastasis)

NCI-H727-Lung carcinoid
0.0
SU86.86-Pancreatic carcinoma
0.0

(liver metastasis)

NCI-UMC-11-Lung
47.3
BxPC-3-Pancreatic
0.0

carcinoid

adenocarcinoma

LX-1-Small cell lung cancer
0.0
HPAC-Pancreatic adenocarcinoma
0.0

Colo-205-Colon cancer
0.0
MIA PaCa-2-Pancreatic carcinoma
0.0

KM12-Colon cancer
0.0
CFPAC-1-Pancreatic ductal
0.0

adenocarcinoma

KM20L2-Colon cancer
0.0
PANC-1-Pancreatic epithelioid
0.0

ductal carcinoma

NCI-H716-Colon cancer
0.0
T24-Bladder carcinma (transitional
0.0

cell)

SW-48-Colon
0.0
5637-Bladder carcinoma
0.0

adenocarcinoma

SW1116-Colon
0.0
HT-1197-Bladder carcinoma
0.0

adenocarcinoma

LS 174T-Colon
0.0
UM-UC-3-Bladder carcinma
0.0

adenocarcinoma

(transitional cell)

SW-948-Colon
0.0
A204-Rhabdomyosarcoma
0.0

adenocarcinoma

SW-480-Colon
0.0
HT-1080-Fibrosarcoma
0.0

adenocarcinoma

NCI-SNU-5-Gastric
0.0
MG-63-Osteosarcoma
3.5

carcinoma

KATO III-Gastric carcinoma
0.0
SK-LMS-1-Leiomyosarcoma
0.0

(vulva)

NCI-SNU-16-Gastric
0.0
SJRH30-Rhabdomyosarcoma (met
0.0

carcinoma

to bone marrow)

NCI-SNU-1-Gastric
0.0
A431-Epidermoid carcinoma
0.0

carcinoma

RF-1-Gastric
0.0
WM266-4-Melanoma
5.6

adenocarcinoma

RF-48-Gastric
0.0
DU 145-Prostate carcinoma (brain
0.0

adenocarcinoma

metastasis)

MKN-45-Gastric carcinoma
0.0
MDA-MB-468-Breast
0.0

adenocarcinoma

NCI-N87-Gastric carcinoma
7.0
SCC-4-Squamous cell carcinoma
0.0

of tongue

OVCAR-5-Ovarian
0.0
SCC-9-Squamous cell carcinoma
0.0

carcinoma

of tongue

RL95-2-Uterine carcinoma
0.0
SCC-15-Squamous cell carcinoma
0.0

of tongue

HelaS3-Cervical
2.7
CAL 27-Squamous cell carcinoma
0.0

adenocarcinoma

of tongue

[0460]

31

TABLE GF

Panel 4D

Rel. Exp. (%) Ag2355,

Rel. Exp. (%) Ag2355,

Tissue Name
Run 164038075
Tissue Name
Run 164038075

Secondary Th1 act
0.0
HUVEC IL-1beta
0.0

Secondary Th2 act
0.0
HUVEC IFN gamma
0.0

Secondary Tr1 act
0.0
HUVEC TNF alpha + IFN
0.0

gamma

Secondary Th1 rest
0.0
HUVEC TNF alpha + IL4
0.0

Secondary Th2 rest
0.0
HUVEC IL-11
0.0

Secondary Tr1 rest
0.0
Lung Microvascular EC none
0.0

Primary Th1 act
0.0
Lung Microvascular EC
0.0

TNF alpha + IL-1beta

Primary Th2 act
0.0
Microvascular Dermal EC none
0.0

Primary Tr1 act
0.0
Microvascular Dermal EC
0.0

TNF alpha + IL-1beta

Primary Th1 rest
0.0
Bronchial epithelium TNF alpha +
0.0

IL1beta

Primary Th2 rest
0.0
Small airway epithelium none
0.0

Primary Tr1 rest
0.0
Small airway epithelium
0.0

TNF alpha + IL-1beta

CD45RA CD4 lymphocyte
0.0
Coronery artery SMC rest
0.0

act

CD45RO CD4 lymphocyte
0.0
Coronery artery SMC TNF alpha +
14.8

act

IL-1beta

CD8 lymphocyte act
0.0
Astrocytes rest
0.0

Secondary CD8
0.0
Astrocytes TNF alpha + IL-1beta
0.0

lymphocyte rest

Secondary CD8
0.0
KU-812 (Basophil) rest
0.0

lymphocyte act

CD4 lymphocyte none
0.0
KU-812 (Basophil)
22.4

PMA/ionomycin

2ry Th1/Th2/Tr1_anti-
0.0
CCD1106 (Keratinocytes) none
0.0

CD95 CH11

LAK cells rest
0.0
CCD1106 (Keratinocytes)
0.0

TNF alpha + IL-1beta

LAK cells IL-2
0.0
Liver cirrhosis
71.2

LAK cells IL-2 + IL-12
0.0
Lupus kidney
0.0

LAK cells IL-2 + IFN
0.0
NCI-H292 none
0.0

gamma

LAK cells IL-2 + IL-18
0.0
NCI-H292 IL-4
0.0

LAK cells
0.0
NCI-H292 IL-9
0.0

PMA/ionomycin

NK Cells IL-2 rest
0.0
NCI-H292 IL-13
0.0

Two Way MLR 3 day
0.0
NCI-H292 IFN gamma
0.0

Two Way MLR 5 day
0.0
HPAEC none
0.0

Two Way MLR 7 day
0.0
HPAEC TNF alpha + IL-1beta
0.0

PBMC rest
0.0
Lung fibroblast none
34.2

PBMC PWM
0.0
Lung fibroblast TNF alpha + IL-
0.0

1beta

PBMC PHA-L
0.0
Lung fibroblast IL-4
0.0

Ramos (B cell) none
0.0
Lung fibroblast IL-9
0.0

Ramos (B cell) ionomycin
0.0
Lung fibroblast IL-13
0.0

B lymphocytes PWM
0.0
Lung fibroblast IFN gamma
13.0

B lymphocytes CD40L
0.0
Dermal fibroblast CCD 1070 rest
0.0

and IL-4

EOL-1 dbcAMP
0.0
Dermal fibroblast CCD1070
0.0

TNF alpha

EOL-1 dbcAMP
0.0
Dermal fibroblast CCD 1070 IL-
0.0

PMA/ionomycin

1beta

Dendritic cells none
0.0
Dermal fibroblast IFN gamma
0.0

Dendritic cells LPS
0.0
Dermal fibroblast IL-4
0.0

Dendritic cells anti-CD40
46.0
IBD Colitis 2
12.4

Monocytes rest
0.0
IBD Crohn's
16.6

Monocytes LPS
0.0
Colon
100.0

Macrophages rest
35.6
Lung
42.9

Macrophages LPS
0.0
Thymus
0.0

HUVEC none
0.0
Kidney
0.0

HUVEC starved
0.0

[0461] Panel 1.3D Summary: Ag2355 The expression of the CG54335-01 gene is low but significant in two breast cancer cell lines. Of note is the fact that the two breast cancer cell lines that express this gene are estrogen receptor positive. Thus, expression of this gene may be indicative of estrogen receptor status on breast cancer cells and may have implications to breast cancer cell biology. In addition, therapeutic modulation of this gene may have utility in the treatment of breast cancer or other breast disease. Ag1635 The expression of gene CG54335-01 is low/undetectable (CT values>35) in all the tissues on this panel.

[0462] Panel 2.2 Summary: Ag1635 Expression of gene CG54335-01 on this panel is too low to be reliable (Ct values>35).

[0463] Panel 2D Summary: Ag2355 Expression of the CG54335-01 gene is highest in a sample derived from an ovarian cancer. Samples in which there is also expression are many fold lower than the ovarian cancer. Thus, this gene may be useful for the diagnosis or therapeutic intervention for ovarian cancer.

[0464] Panel 3D Summary: Ag2355 The expression of the CG54335-01 gene in panel 3D appears to be associated with lung cancer cell lines. Furthermore, the cell line that expresses this gene in most abundance is neuroendocrine in origin. Neuroendocrine tumors are very unique and thus, the CG54335-01 gene may represent a unique marker of this type of cancer. In addition, therapeutic modulation of this gene may be useful for the treatment of neuroendocrine tumors in the lung.

[0465] Panel 4D Summary: Ag2355 The CG54335-01 transcript is expressed in normal colon but not in colons from patients with Crohn's disease or colitis. Protein therapeutics designed with the putative GPCR encoded for by this gene could be used reduce or eliminate inflammation and tissue destruction due to IBD. Ag1635 Expression is low/undetectable (CTs>35) across all of the samples on this panel.

[0466] H. CG55993-03/GMAC011711_G: Olfactory Receptor

[0467] Expression of gene CG55993-03 was assessed using the primer-probe sets Ag2322, Ag2365 and Ag2350, described in Tables HA, HB and HC. Results of the RTQ-PCR runs are shown in Tables HD, HE, HF, and HG.

32TABLE HAProbe Name Ag2322StartSEQ IDPrimersSequencesLengthPositionNO:Forward5′-atcccactgtgcttcatgtatc-3′22132192ProbeTET-5′-atcccgggcaactgcacaattctttt-3′-TAMRA26162193Reverse5′-agtgagcgctctgttttaatga-3′22190194

[0468]

33

TABLE HB

Probe Name Ag2365

Start
SEQ ID

Primers
Seqences
Length
Position
NO:

Forward
5′-atcccactgtgcttcatgtatc-3′
22
132
198

Probe
TET-5′-atcccgggcaactgcacaattctttt-3′TAMRA
26
162
196

Reverse
5′-agtgagcgctctgttttaatga-3′
22
190
197

[0469]

34

TABLE HC

Probe Name Ag2350

Start
SEQ ID

Primers
Sequences
Length
Position
NO:

Forward
5′-atcccactgtgcttcatgtatc-3′
22
132
198

Probe
TET-5′-atcccgggcaactgcacaattctttt-3′-TAMRA
26
162
199

Reverse
5′-agtgagcgctctgttttaatga-3′
22
190
200

[0470]

35

TABLE HD

AI_comprehensive panel_v1.0

Rel. Exp. (%) Ag2322,

Rel. Exp. (%) Ag2322,

Tissue Name
Run 229743870
Tissue Name
Run 229743870

110967 COPD-F
2.2
112427 Match Control
0.0

Psoriasis-F

110980 COPD-F
0.0
112418 Psoriasis-M
0.0

110968 COPD-M
0.0
112723 Match Control
1.5

Psoriasis-M

110977 COPD-M
0.0
112419 Psoriasis-M
0.0

110989 Emphysema-F
0.0
112424 Match Control
3.0

Psoriasis-M

110992 Emphysema-F
0.0
112420 Psoriasis-M
0.0

110993 Emphysema-F
0.0
112425 Match Control
2.3

Psoriasis-M

110994 Emphysema-F
0.0
104689 (MF) OA Bone-
0.0

Backus

110995 Emphysema-F
0.0
104690 (MF) Adj “Normal”
0.0

Bone-Backus

110996 Emphysema-F
1.7
104691 (MF) OA
0.0

Synovium-Backus

110997 Asthma-M
2.5
104692 (BA) OA Cartilage-
0.0

Backus

111001 Asthma-F
0.0
104694 (BA) OA Bone-
0.0

Backus

111002 Asthma-F
0.0
104695 (BA) Adj “Normal”
0.0

Bone-Backus

111003 Atopic Asthma-F
0.0
104696 (BA) OA Synovium-
0.0

Backus

111004 Atopic Asthma-F
0.0
104700 (SS) OA Bone-
0.0

Backus

111005 Atopic Asthma-F
0.0
104701 (SS) Adj “Normal”
0.0

Bone-Backus

111006 Atopic Asthma-F
0.0
104702 (SS) OA Synovium-
0.0

Backus

111417 Allergy-M
0.0
117093 OA Cartilage Rep7
0.0

112347 Allergy-M
4.6
112672 OA Bone5
0.0

112349 Normal Lung-F
4.5
112673 OA Synovium5
0.0

112357 Normal Lung-F
2.0
112674 OA Synovial Fluid
0.0

cells5

112354 Normal Lung-M
0.0
117100 OA Cartilage Rep14
0.0

112374 Crohns-F
0.0
112756 OA Bone9
100.0

112389 Match Control
0.0
112757 OA Synovium9
0.0

Crohns-F

112375 Crohns-F
0.0
112758 OA Synovial Fluid
0.0

Cells9

112732 Match Control
0.0
117125 RA Cartilage Rep2
0.0

Crohns-F

112725 Crohns-M
0.0
113492 Bone2 RA
1.7

112387 Match Control
2.3
113493 Synovium2 RA
0.0

Crohns-M

112378 Crohns-M
11.3
113494 Syn Fluid Cells RA
0.0

112390 Match Control
0.0
113499 Cartilage4 RA
0.0

Crohns-M

112726 Crohns-M
2.6
113500 Bone4 RA
0.0

112731 Match Control
0.0
113501 Synovium4 RA
0.0

Crohns-M

112380 Ulcer Col-F
0.0
113502 Syn Fluid Cells4 RA
0.0

112734 Match Control
0.0
113495 Cartilage3 RA
0.0

Ulcer Col-F

112384 Ulcer Col-F
1.9
113496 Bone3 RA
0.0

112737 Match Control
0.0
113497 Synovium3 RA
0.0

Ulcer Col-F

112386 Ulcer Col-F
0.0
113498 Syn Fluid Cells3 RA
0.0

112738 Match Control
0.0
117106 Normal Cartilage
0.0

Ulcer Col-F

Rep20

112381 Ulcer Col-M
0.0
113663 Bone3 Normal
2.6

112735 Match Control
0.0
113664 Synovium3 Normal
2.6

Ulcer Col-M

112382 Ulcer Col-M
0.0
113665 Syn Fluid Cells3
0.0

Normal

112394 Match Control
2.1
117107 Normal Cartilage
0.0

Ulcer Col-M

Rep22

112383 Ulcer Col-M
1.4
113667 Bone4 Normal
0.0

112736 Match Control
0.0
113668 Synovium4 Normal
2.2

Ulcer Col-M

112423 Psoriasis-F
0.0
113669 Syn Fluid Cells4
2.0

Normal

[0471]

36

TABLE HE

Panel 1.3D

Rel. Exp. (%)
Rel. Exp. (%)

Rel. Exp. (%)
Rel. Exp. (%)

Ag2322, Run
Ag2350, Run

Ag2322, Run
Ag2350, Run

Tissue Name
165627868
165974845
Tissue Name
165627868
165974845

Liver
0.0
0.0
Kidney (fetal)
0.0
0.0

adenocarcinoma

Pancreas
0.0
0.0
Renal ca. 786-0
0.0
0.0

Pancreatic ca.
0.0
0.0
Renal ca. A498
0.0
0.0

CAPAN 2

Adrenal gland
0.0
0.0
Renal ca. RXF
0.0
0.0

393

Thyroid
0.0
0.0
Renal ca. ACHN
0.0
0.0

Salivary gland
0.0
0.0
Renal ca. UO-31
0.0
0.0

Pituitary gland
0.0
7.6
Renal ca. TK-10
0.0
0.0

Brain (fetal)
0.0
0.0
Liver
0.0
0.0

Brain (whole)
0.0
0.0
Liver (fetal)
0.0
0.0

Brain (amygdala)
0.0
0.0
Liver ca.
0.0
0.0

(hepatoblast)

HepG2

Brain (cerebellum)
0.0
0.0
Lung
0.0
0.0

Brain (hippocampus)
0.0
0.0
Lung (fetal)
0.0
0.0

Brain (substantia
0.0
0.0
Lung ca. (small
0.0
0.0

nigra)

cell) LX-1

Brain (thalamus)
0.0
0.0
Lung ca. (small
0.0
31.2

cell) NCI-H69

Cerebral Cortex
0.0
0.0
Lung ca. (s.cell
100.0
100.0

var.) SHP-77

Spinal cord
3.4
0.0
Lung ca. (large
0.0
0.0

cell)NCI-H460

glio/astro U87-MG
0.0
0.0
Lung ca. (non-
0.0
0.0

sm. cell) A549

glio/astro U-118-
0.0
0.0
Lung ca. (non-
0.0
0.0

MG

s.cell) NCI-H23

astrocytoma
0.0
0.0
Lung ca. (non-
0.0
0.0

SW1783

s.cell) HOP-62

neuro*; met SK-N-
0.0
0.0
Lung ca. (non-
0.0
0.0

AS

s.cl) NCI-H522

astrocytoma SF-539
0.0
0.0
Lung ca.
0.0
0.0

(squam.) SW 900

astrocytoma SNB-75
0.0
0.0
Lung ca.
16.6
15.5

(squam.) NCI-

H596

glioma SNB-19
2.5
0.0
Mammary gland
0.0
0.0

glioma U251
0.0
0.0
Breast ca.*
0.0
0.0

(pl.ef) MCF-7

glioma SF-295
0.0
0.0
Breast ca.*
0.0
0.0

(pl.ef) MDA-

MB-231

Heart (fetal)
0.0
0.0
Breast ca.*
0.0
0.0

(pl.ef) T47D

Heart
0.0
0.0
Breast ca. BT-
0.0
0.0

549

Skeletal muscle
0.0
0.0
Breast ca. MDA-N
0.0
0.0

(fetal)

Skeletal muscle
0.0
0.0
Ovary
0.0
0.0

Bone marrow
0.0
0.0
Ovarian ca.
1.7
0.0

OVCAR-3

Thymus
0.0
0.0
Ovarian ca.
0.0
0.0

OVCAR-4

Spleen
0.0
0.0
Ovarian ca.
0.0
0.0

OVCAR-5

Lymph node
0.0
0.0
Ovarian ca.
0.0
4.8

OVCAR-8

Colorectal
1.4
0.0
Ovarian ca.
0.0
0.0

IGROV-1

Stomach
0.0
0.0
Ovarian ca.*
0.0
0.0

(ascites) SK-OV-3

Small intestine
0.0
0.0
Uterus
0.0
0.0

Colon ca. SW480
0.0
0.0
Plancenta
0.0
0.0

Colon ca.*
0.0
0.0
Prostate
0.0
0.0

SW620(SW480 met)

Colon ca. HT29
0.0
0.0
Prostate ca.*
0.0
0.0

(bone met)PC-3

Colon ca. HCT-116
0.0
0.0
Testis
0.0
0.0

Colon ca. CaCo-2
0.0
0.0
Melanoma
0.0
0.0

Hs688(A).T

Colon ca.
0.0
0.0
Melanoma*
0.0
0.0

tissue(ODO3866)

(met)

Hs688(B).T

Colon ca. HCC-2998
0.0
0.0
Melanoma
0.0
0.0

UACC-62

Gastric ca.* (liver
3.3
0.0
Melanoma M14
0.0
0.0

met) NCI-N87

Bladder
0.0
6.9
Melanoma LOX
0.0
0.0

IMVI

Trachea
0.0
0.0
Melanoma*
0.0
0.0

(met) SK-MEL-5

Kidney
0.0
0.0
Adipose
0.0
0.0

[0472]

37

TABLE HF

Panel 2D

Rel. Exp. (%) Ag2350,

Rel. Exp. (%) Ag2350,

Tissue Name
Run 164079723
Tissue Name
Run 164079723

Normal Colon
7.6
Kidney Margin 8120608
0.0

CC Well to Mod Diff
4.1
Kidney Cancer 8120613
0.0

(ODO3866)

CC Margin (ODO3866)
1.5
Kidney Margin 8120614
0.0

CC Gr.2 rectosigmoid
3.0
Kidney Cancer 9010320
0.0

(ODO3868)

CC Margin (ODO3868)
0.0
Kidney Margin 9010321
0.0

CC Mod Diff (ODO3920)
0.0
Normal Uterus
0.0

CC Margin (ODO3920)
7.1
Uterus Cancer 064011
3.7

CC Gr.2 ascend colon
0.0
Normal Thyroid
0.0

(ODO3921)

CC Margin (ODO3921)
0.0
Thyroid Cancer 064010
0.0

CC from Partial Hepatectomy
0.0
Thyroid Cancer A302152
0.0

(ODO4309) Mets

Liver Margin (ODO4309)
0.0
Thyroid Margin A302153
0.0

Colon mets to lung (OD04451-
0.0
Normal Breast
0.0

01)

Lung Margin (OD04451-02)
0.0
Breast Cancer (OD04566)
7.6

Normal Prostate 6546-1
22.4
Breast Cancer (OD04590-
0.0

01)

Prostate Cancer (OD04410)
74.7
Breast Cancer Mets
0.0

(OD04590-03)

Prostate Margin (OD04410)
100.0
Breast Cancer Metastasis
0.0

(OD04655-05)

Prostate Cancer (OD04720-01)
30.1
Breast Cancer 064006
0.0

Prostate Margin (OD04720-02)
38.7
Breast Cancer 1024
0.0

Normal Lung 061010
0.0
Breast Cancer 9100266
0.0

Lung Met to Muscle
0.0
Breast Margin 9100265
1.7

(ODO4286)

Muscle Margin (ODO4286)
0.0
Breast Cancer A209073
7.2

Lung Malignant Cancer
25.5
Breast Margin A2090734
0.0

(OD03126)

Lung Margin (OD03126)
0.0
Normal Liver
0.0

Lung Cancer (OD04404)
6.5
Liver Cancer 064003
0.0

Lung Margin (OD04404)
0.0
Liver Cancer 1025
0.0

Lung Cancer (OD04565)
0.0
Liver Cancer 1026
0.0

Lung Margin (OD04565)
0.0
Liver Cancer 6004-T
0.0

Lung Cancer (OD04237-01)
0.0
Liver Tissue 6004-N
0.0

Lung Margin (OD04237-02)
0.0
Liver Cancer 6005-T
0.0

Ocular Mel Met to Liver
0.0
Liver Tissue 6005-N
0.0

(ODO4310)

Liver Margin (ODO4310)
0.0
Normal Bladder
0.0

Melanoma Mets to Lung
0.0
Bladder Cancer 1023
0.0

(OD04321)

Lung Margin (OD04321)
0.0
Bladder Cancer A302173
6.1

Normal Kidney
0.0
Bladder Cancer
0.0

(OD04718-01)

Kidney Ca, Nuclear grade 2
0.0
Bladder Normal Adjacent
0.0

(OD04338)

(OD04718-03)

Kidney Margin (OD04338)
3.6
Normal Ovary
0.0

Kidney Ca Nuclear grade 1/2
0.0
Ovarian Cancer 064008
0.0

(OD04339)

Kidney Margin (OD04339)
0.0
Ovarian Cancer
0.0

(OD04768-07)

Kidney Ca, Clear cell type
0.0
Ovary Margin (OD04768-
0.0

(OD04340)

08)

Kidney Margin (OD04340)
0.0
Normal Stomach
0.0

Kidney Ca, Nuclear grade 3
1.7
Gastric Cancer 9060358
0.0

(OD04348)

Kidney Margin (OD04348)
0.0
Stomach Margin 9060359
0.0

Kidney Cancer (OD04622-01)
0.0
Gastric Cancer 9060395
0.0

Kidney Margin (OD04622-03)
0.0
Stomach Margin 9060394
0.0

Kidney Cancer (OD04450-01)
0.0
Gastric Cancer 9060397
0.0

Kidney Margin (OD04450-03)
0.0
Stomach Margin 9060396
0.0

Kidney Cancer 8120607
0.0
Gastric Cancer 064005
3.7

[0473]

38

TABLE HG

Panel 4D

Rel. Exp. (%)
Rel. Exp. (%)

Rel. Exp. (%)
Rel. Exp. (%)

Ag2322, Run
Ag2350, Run

Ag2322, Run
Ag2350, Run

Tissue Name
162360932
164145590
Tissue Name
162360932
164145590

Secondary Th1 act
0.0
0.0
HUVEC IL-1beta
0.0
0.0

Secondary Th2 act
0.0
0.0
HUVEC IFN gamma
0.0
0.0

Secondary Tr1 act
0.0
29.3
HUVEC TNF alpha +
0.0
0.0

IFN gamma

Secondary Th1 rest
0.0
0.0
HUVEC TNF alpha +
0.0
0.0

IL4

Secondary Th2 rest
0.0
0.0
HUVEC IL-11
0.0
0.0

Secondary Tr1 rest
0.0
0.0
Long Microvascular
0.0
0.0

EC none

Primary Th1 act
0.0
0.0
Lung Microvascular
0.0
0.0

EC TNF alpha + IL-

1beta

Primary Th2 act
0.0
0.0
Microvascular
0.0
0.0

Dermal EC none

Primary Tr1 act
0.0
0.0
Microsvasular Dermal
0.0
0.0

EC TNF alpha + IL-

1beta

Primary Th1 rest
0.0
0.0
Bronchial epithelium
0.0
0.0

TNF alpha + IL1beta

Primary Th2 rest
0.0
0.0
Small airway
0.0
0.0

epithelium none

Primary Tr1 rest
0.0
0.0
Small airway
0.0
0.0

epithelium TNF alpha +

IL-1beta

CD45RA CD4
0.0
0.0
Coronery artery SMC
0.0
0.0

lymphocyte act

rest

CD45RO CD4
0.0
0.0
Coronery artery SMC
0.0
48.0

lymphocyte act

TNF alpha + IL-1beta

CD8 lymphocyte act
0.0
23.3
Astrocytes rest
0.0
0.0

Secondary CD8
0.0
0.0
Astrocytes TNF alpha +
0.0
0.0

lymphocyte rest

IL-1beta

Secondary CD8
21.2
0.0
KU-812 (Basophil)
0.0
0.0

lymphocyte act

rest

CD4 lymphocyte
0.0
0.0
KU-812 (Basophil)
0.0
0.0

none

PMA/ionomycin

2ry
0.0
0.0
CCD1106
0.0
0.0

Th1/Th2/Tr1_anti-

(Keratinocytes) none

CD95 CH11

LAK cells rest
0.0
0.0
CCD1106
0.0
0.0

(Keratinocytes)

TNF alpha + IL-1beta

LAK cells IL-2
0.0
0.0
Liver cirrhosis
100.0
100.0

LAK cells IL-2 + IL-
0.0
0.0
Lupus kidney
0.0
0.0

12

LAK cells IL-2 + IFN
0.0
0.0
NCI-H292 none
0.0
0.0

gamma

LAK cells IL-2 + IL-
0.0
0.0
NCI-H292 IL-4
0.0
0.0

18

LAK cells
0.0
0.0
NCI-H292 IL-9
0.0
0.0

PMA/ionomycin

NK Cells IL-2 rest
0.0
0.0
NCI-H292 IL-13
0.0
0.0

Two Way MLR 3
0.0
0.0
NCI-H292 IFN
0.0
0.0

day

gamma

Two Way MLR 5
0.0
0.0
HPAEC none
0.0
0.0

day

Two Way MLR 7
0.0
0.0
HPAEC TNF alpha +
0.0
0.0

day

IL-1beta

PBMC rest
0.0
0.0
Lung fibroblast none
0.0
21.0

PBMC PWM
0.0
0.0
Lung fibroblast TNF
0.0
0.0

alpha + IL-1beta

PBMC PHA-L
0.0
0.0
Lung fibroblast IL-4
0.0
0.0

Ramos (B cell) none
0.0
0.0
Lung fibroblast IL-9
0.0
0.0

Ramos (B cell)
0.0
0.0
Lung fibroblast IL-13
0.0
0.0

ionomycin

B lymphocytes PWM
7.1
0.0
Lung fibroblast IFN
0.0
40.1

gamma

B lymphocytes
0.0
0.0
Dermal fibroblast
0.0
0.0

CD40L and IL-4

CCD1070 rest

EOL-1 dbcAMP
0.0
0.0
Dermal fibroblast
0.0
0.0

CCD1070 TNF alpha

EOL-1 dbcAMP
0.0
0.0
Dermal fibroblast
0.0
0.0

PMA/ionomycin

CCD1070 IL-1beta

Dendritic cells none
0.0
0.0
Dermal fibroblast IFN
0.0
0.0

gamma

Dendritic cells LPS
0.0
0.0
Dermal fibroblast IL-4
0.0
0.0

Dendritic cells anti-
0.0
0.0
IBD Colitis 2
43.2
19.2

CD40

Monocytes rest
0.0
0.0
IBD Crohn's
0.0
15.3

Monocytes LPS
0.0
0.0
Colon
0.0
0.0

Macrophages rest
0.0
0.0
Lung
0.0
0.0

Macrophages LPS
0.0
0.0
Thymus
0.0
19.1

HUVEC none
0.0
0.0
Kidney
0.0
0.0

HUVEC starved
0.0
0.0

[0474] AI_comprehensive panel_v1.0 Summary: Ag2322 Low but significant expression of the CG55993-03 gene is detected in an osteoarthritic bone sample(CT=33). The protein encoded by this gene is homologous to a G-protein coupled receptors. These receptors have been shown to be involved in calcium homeostasis and control (reference 1). Changes in bone tissue (eg subchondral bone hardening) are identified in osteoarthritis. Therefore, therapeutic modulation of this gene product may ameliorate non-homeostatic bone-related changes found in osteoarthritis (Gardella and Juppner, Molecular properties of the PTH/PTHrP receptor. Trends Endocrinol Metab 12(5):210-7, 2001)

[0475] CNS_neurodegeneration_v1.0 Summary: Ag2365 Data from one experiment with this probe and primer set and the CG55993-03 gene is not included due to a probable probe failure.

[0476] Panel 1.3D Summary: Ag2322/2350 In two runs with the same probe and primer set, highest expression of the CG55993-03 gene is seen in a sample derived from a small cell lung cancer cell line (SHP-77) (CTs=32-33). There is apparent expression in other small cell lung cancer cell lines as well. Thus, the expression of this gene could be used to distinguish SHP-77 cells and other small cell lung cancer cell lines from other samples on the panel. Moreover, therapeutic modulation of this gene, through the use of small molecule drugs, antibodies or protein therapeutics might be of use in the treatment of small cell lung cancer. Please note that a third experiment with the probe and primer Ag2365 showed low/undetectable expression in all samples on this panel (CTs>35). (Data not shown.)

[0477] Panel 2D Summary: Ag2350 The expression of the CG55993-03 gene is highest in a sample derived from normal prostate tissue adjacent to a prostate cancer (CT=32.3). This pattern holds for another matched pair of prostate cancer and normal tissues. There is low to no expression in other tissues, therefore, this gene appears to show prostate specific expression. Thus, the expression of this gene could be used to distinguish prostate derived tissues from other tissues in the panel. Moreover, therapeutic modulation of this this gene, through the use of small molecule drugs, antibodies or protein therapeutics might be of benefit in the treatment of prostate cancer.

[0478] Panel 3D Summary: Ag2365 Data from one experiment with this probe and primer set and the CG55993-03 gene is not included due to a probable probe failure.

[0479] Panel 4D Summary: Ag2322/Ag2350 The CG55993-03 transcript is only expressed at significant levels in liver cirrhosis. The transcript is not expressed in normal liver in panel 2 or 1. The transcript or the protein it encodes could be used for detection of liver cirrhosis. The putative GPCR encoded for by this transcript may also play an important role in liver cirrhosis. Therapeutics designed with the protein encoded for by this transcript could be important for maintaining or restoring normal function to the liver undergoing cirrhosis. Please note that a third experiment with the probe and primer Ag2365 showed low/undetectable expression in all samples on this panel (CTs>35). (Data not shown.)

[0480] I. CG55978-01/GMAC011711_F: Olfactory Receptor

[0481] Expression of gene CG55978-01 was assessed using the primer-probe set Ag2349, described in Table IA. Results of the RTQ-PCR runs are shown in Tables IB, IC, ID, and IE.

39TABLE IAProbe Name Ag2349StartSEQ IDPrimersSequencesLengthPositionNO:Forward5′-tgtccttctgcagttctatggt-3′22512201ProbeTET-5′-tactgctaccatgttgatctcatcca-3′-TAMRA26547202Reverse5′-cctattgtctgtgcaggagagt-3′22573203

[0482]

40

TABLE IB

Panel 1.3D

Rel. Exp. (%)
Rel. Exp. (%)

Rel. Exp. (%)
Rel. Exp. (%)

Ag2349, Run
Ag2349, Run

Ag2349, Run
Ag2349, Run

Tissue Name
160653021
165632362
Tissue Name
160653021
165632362

Liver
0.0
0.0
Kidney (fetal)
0.6
7.3

adenocarcinoma

Pancreas
0.0
0.0
Renal ca. 786-0
0.0
0.0

Pancreatic ca.
0.0
0.0
Renal ca. A498
0.3
0.0

CAPAN 2

Adrenal gland
0.0
0.0
Renal ca. RXF
0.0
0.0

393

Thyroid
0.0
0.0
Renal ca. ACHN
0.0
0.0

Salivary gland
0.0
0.0
Renal ca. UO-31
0.0
0.0

Pituitary gland
0.0
0.0
Renal ca. TK-10
0.0
0.0

Brain (fetal)
0.0
0.0
Liver
0.0
0.0

Brain (whole)
0.0
1.8
Liver (fetal)
0.0
0.0

Brain (amygdala)
0.0
0.0
Liver ca.
0.0
0.0

(hepatoblast)

HepG2

Brain (cerebellum)
0.0
0.0
Lung
0.0
0.0

Brain
0.0
0.0
Lung (fetal)
0.4
0.0

(hippocampus)

Brain (substantia
0.0
0.0
Lung ca. (small
0.0
0.0

nigra)

cell) LX-1

Brain (thalamus)
0.0
0.0
Lung ca. (small
13.0
4.0

cell) NCI-H69

Cerebral Cortex
0.8
0.0
Lung ca. (s.cell
100.0
100.0

var.) SHP-77

Spinal cord
0.0
0.0
Lung ca. (large
0.0
0.0

cell)NCI-H460

glio/astro U87-MG
0.0
0.0
Lung ca. (non-
1.0
0.0

sm. cell) A549

glio/astro U-118-
0.9
4.0
Lung ca. (non-
0.0
0.0

MG

s.cell) NCI-H23

astrocytoma
0.0
0.0
Lung ca. (non-
0.0
5.0

SW1783

s.cell) HOP-62

neuro*; met SK-N-
0.0
0.0
Lung ca. (non-
0.0
0.0

AS

s.cl) NCI-H522

astrocytoma SF-539
0.0
0.0
Lung ca.
0.0
0.0

(squam.) SW 900

astrocytoma SNB-
0.0
0.0
Lung ca.
6.6
20.2

75

(squam.) NCI-

H596

glioma SNB-19
0.0
0.0
Mammary gland
0.0
0.0

glioma U251
0.0
0.0
Breast ca.*
0.0
0.0

(pl.ef) MCF-7

glioma SF-295
0.6
0.0
Breast ca.*
0.0
0.0

(pl.ef) MDA-

MB-231

Heart (Fetal)
0.0
0.0
Breast ca.* (pl.
0.0
0.0

ef) T47D

Heart
0.0
2.3
Breat ca.* BT-
0.7
0.0

549

Skeletal muscle
1.0
0.0
Breast ca. MDA-N
0.6
0.0

(Fetal)

Skeletal muscle
0.0
4.2
Ovary
0.0
4.5

Bone marrow
0.0
0.0
Ovarian ca.
0.0
0.0

OVCAR-3

Thymus
0.0
0.0
Ovarian ca.
0.0
0.0

OVCAR-4

Spleen
0.0
0.0
Ovarian ca.
0.0
0.0

OVCAR-5

Lymph node
0.0
0.0
Ovarian ca.
0.0
0.0

OVCAR-8

Colorectal
2.7
6.8
Ovarian ca.
0.0
1.9

IGROV-1

Stomach
0.0
2.2
Ovarian ca.
0.0
3.4

(ascites) SK-OV-3

Small intestine
0.0
0.0
Uterus
0.0
0.0

Colon ca. SW480
3.2
2.2
Placenta
0.0
0.0

Colon ca.* SW620
0.0
0.0
Prostate
0.6
3.0

(SW480 met)

Colon ca. HT29
0.5
0.0
Prostate ca.*
0.5
0.0

(bone met) PC-3

Colon ca. HCT-116
0.0
2.3
Testis
0.0
0.0

Colon ca. CaCo-2
0.0
0.0
Melanoma
0.0
0.0

Hs688(A).T

CC Well to Mod
0.0
0.0
Melanoma*
0.0
0.0

Diff (ODO3866)

(met)

Hs688(B).T

Colon ca. HCC-
0.0
0.0
Melanoma
0.0
0.0

2998

UACC-62

Gastric ca. (liver
0.0
4.3
Melanoma M14
0.0
0.0

met) NCI-N87

Bladder
0.0
0.0
Melanoma LOX
0.0
0.0

IMVI

Trachea
0.0
0.0
Melanoma*
0.0
0.0

(met) SK-MEL-5

Kidney
0.0
0.0
Adipose
0.5
0.0

[0483]

41

TABLE IC

Panel 2D

Rel. Exp. (%) Ag2349,

Rel. Exp. (%) Ag2349,

Tissue Name
Run 160653160
Tissue Name
Run 160653160

Normal Colon
7.1
Kidney Margin 8120608
0.0

CC Well to Mod Diff
0.0
Kidney Cancer 8120613
1.1

(ODO3866)

CC Margin (ODO3866)
1.3
Kidney Margin 8120614
0.0

CC Gr.2 rectosigmoid
0.0
Kidney Cancer 9010320
0.0

(ODO3868)

CC Margin (ODO3868)
0.6
Kidney Margin 9010321
0.0

CC Mod Diff (ODO3920)
1.1
Normal Uterus
0.0

CC Margin (ODO3920)
1.6
Uterine Cancer 064011
0.0

CC Gr.2 ascend colon
0.0
Normal Thyroid
0.0

(ODO3921)

CC Margin (ODO3921)
2.5
Thyroid Cancer
0.0

CC from Partial Hepatectomy
0.0
Thyroid Cancer A302152
0.0

(ODO4309) Mets

Liver Margin (ODO4309)
0.0
Thyroid Margin A302153
0.0

Colon mets to lung (OD04451-
0.0
Normal Breast
0.0

01)

Lung Margin (OD04451-02)
0.0
Breast Cancer
0.0

Normal Prostate 6546-1
9.8
Breast Cancer (OD04590-
0.0

01)

Prostate Cancer (OD04410)
100.0
Breast Cancer Mets
0.0

(OD04590-03)

Prostate Margin (OD04410)
77.9
Breast Cancer Metastasis
0.0

Prostate Cancer (OD04720-01)
22.5
Breast Cancer
0.0

Prostate Margin (OD04720-02)
72.7
Breast Cancer
0.0

Normal Lung
0.0
Breast Cancer 9100266
4.0

Lung Met to Muscle
0.0
Breast Margin 9100265
0.0

(ODO4286)

Muscle Margin (ODO4286)
1.3
Breast Cancer A209073
5.0

Lung Malignant Cancer
17.6
Breast Margin A2090734
0.0

(OD03126)

Lung Margin (OD03126)
1.7
Normal Liver
0.0

Lung Cancer (OD04404)
0.0
Liver Cancer
0.0

Lung Margin (OD04404)
1.2
Liver Cancer 1025
0.0

Lung Cancer (OD04565)
0.0
Liver Cancer 1026
0.0

Lung Margin (OD04565)
0.0
Liver Cancer 6004-T
1.1

Lung Cancer (OD04237-01)
0.0
Liver Tissue 6004-N
0.0

Lung Margin (OD04237-02)
0.0
Liver Cancer 6005-T
0.0

Ocular Mel Met to Liver
0.0
Liver Tissue 6005-N
0.0

(ODO4310)

Liver Margin (ODO4310)
0.0
Normal Bladder
1.8

Melanoma Metastasis
0.0
Bladder Cancer
0.0

Lung Margin (OD04321)
0.0
Bladder Cancer
4.3

Normal Kidney
2.5
Bladder Cancer
0.0

(OD04718-01)

Kidney Ca, Nuclear grade 2
0.0
Bladder Normal Adjacent
0.0

(OD04338)

(OD04718-03)

Kidney Margin (OD04338)
1.1
Normal Ovary
0.5

Kidney Ca Nuclear grade 1/2
0.0
Ovarian Cancer
0.0

(OD04339)

Kidney Margin (OD04339)
0.7
Ovarian Cancer
0.0

(OD04768-07)

Kidney Ca, Clear cell type
0.0
Ovary Margin (OD04768-
0.0

(OD04340)

08)

Kidney Margin (OD04340)
0.0
Normal Stomach
1.3

Kidney Ca, Nuclear grade 3
0.0
Gastric Cancer 9060358
0.0

(OD04348)

Kidney Margin (OD04348)
0.0
Stomach Margin 9060359
0.0

Kidney Cancer (OD04622-01)
0.0
Gastric Cancer 9060395
0.0

Kidney Margin (OD04622-03)
0.0
Stomach Margin 9060394
4.4

Kidney Cancer (OD04450-01)
0.0
Gastric Cancer 9060397
0.0

Kidney Margin (OD04450-03)
0.0
Stomach Margin 9060396
0.0

Kidney Cancer 8120607
0.0
Gastric Cancer 064005
1.1

[0484]

42

TABLE ID

Panel 3D

Rel. Exp. (%)

Rel. Exp. (%)

Ag2349, Run

Ag2349, Run

Tissue Name
164843782
Tissue Name
164843782

Daoy-Medulloblastoma
0.0
Ca Ski-Cervical epidermoid
0.0

carcinoma (metastasis)

TE671-Medulloblastoma
0.0
ES-2-Ovarian clear cell carcinoma
0.0

D283 Med-Medulloblastoma
0.0
Ramos-Stimulated with
0.0

PMA/ionomycin 6 h

PFSK-1-Primitive
0.9
Ramos-Stimulated with
0.0

Neuroectodermal

PMA/ionomycin 14 h

XF-498-CNS
0.0
MEG-01-Chronic myelogenous
0.0

leukemia (megokaryoblast)

SNB-78-Glioma
0.0
Raji-Burkitt's lymphoma
0.0

SF-268-Glioblastoma
0.0
Daudi-Burkitt's lymphoma
0.0

T98G-Glioblastoma
0.0
U266-B-cell plasmacytoma
0.0

SK-N-SH-Neuroblastoma
0.0
CA46-Burkitt's lymphoma
0.0

(metastasis)

SF-295-Glioblastoma
0.0
RL-non-Hodgkin's B-cell
0.0

lymphoma

Cerebellum
0.0
JM1-pre-B-cell lymphoma
0.0

Cerebellum
0.0
Jurkat-T cell leukemia
0.0

NCI-H292-Mucoepidermoid
0.0
TF-1-Erythroleukemia
0.0

lung carcinoma

DMS-114-Small cell lung
0.0
HUT 78-T-cell lymphoma
0.0

cancer

DMS-79-Small cell lung
100.0
U937-Histiocytic lymphoma
0.0

cancer

NCI-H146-Small cell lung
6.7
KU-812-Myelogenous leukemia
0.0

cancer

NCI-H526-Small cell lung
0.0
769-P-Clear cell renal carcinoma
0.0

cancer

NCI-N417-Small cell lung
0.0
Caki-2-Clear cell renal carcinoma
0.0

cancer

NCI-H82-Small cell lung
0.0
SW 839-Clear cell renal carcinoma
0.0

cancer

NCI-H157-Squamous cell
0.0
G401-Wilms' tumor
0.0

lung cancer (metastasis)

NCI-H1155-Large cell lung
0.0
Hs766T-Pancreatic carcinoma (LN
0.0

cancer

metastasis)

NCI-H1299-Large cell lung
0.0
CAPAN-1-Pancreatic
0.0

cancer

adenocarcinoma (liver metastasis)

NCI-H727-Lung carcinoid
0.0
SU86.86-Pancreatic carcinoma
0.0

(liver metastasis)

NCI-UMC-11-Lung
23.2
BxPC-3-Pancreatic
0.0

carcinoid

adenocarcinoma

LX-1-Small cell lung cancer
0.0
HPAC-Pancreatic adenocarcinoma
0.0

Colo-205-Colon cancer
0.0
MIA PaCa-2-Pancreatic carcinoma
0.0

KM12-Colon cancer
0.0
CFPAC-1-Pancreatic ductal
0.0

adenocarcinoma

KM20L2-Colon cancer
0.0
PANC-1-Pancreatic epithelioid
0.0

ductal carcinoma

NCI-H716-Colon cancer
0.0
T24-Bladder carcinma (transitional
0.0

cell)

SW-48-Colon
0.0
5637-Bladder carcinoma
0.0

adenocarcinoma

SW1116-Colon
0.0
HT-1197-Bladder carcinoma
0.0

adenocarcinoma

LS 174T-Colon
0.0
UM-UC-3-Bladder carcinma
0.0

adenocarcinoma

(transitional cell)

SW-948-Colon
0.0
A204-Rhabdomyosarcoma
0.0

adenocarcinoma

SW-480-Colon
0.0
HT-1080-Fibrosarcoma
0.0

adenocarcinoma

NCI-SNU-5-Gastric
0.0
MG-63-Osteosarcoma
0.0

carcinoma

KATO III-Gastric carcinoma
0.0
SK-LMS-1-Leiomyosarcoma
0.0

(vulva)

NCI-SNU-16-Gastric
0.0
SJRH30-Rhabdomyosarcoma (met
0.0

carcinoma

to bone marrow)

NCI-SNU-1-Gastric
0.0
A431-Epidermoid carcinoma
0.0

carcinoma

RF-1-Gastric
0.0
WM266-4-Melanoma
0.0

adenocarcinoma

RF-48-Gastric
0.0
DU 145-Prostate carcinoma (brain
0.0

adenocarcinoma

metastasis)

MKN-45-Gastric carcinoma
0.0
MDA-MB-468-Breast
0.0

adenocarcinoma

NCI-N87-Gastric carcinoma
0.0
SCC-4-Squamous cell carcinoma
0.0

of tongue

OVCAR-5-Ovarian
0.8
SCC-9-Squamous cell carcinoma
0.0

carcinoma

of tongue

RL95-2-Uterine carcinoma
0.0
SCC-15-Squamous cell carcinoma
0.0

of tongue

HelaS3-Cervical
0.0
CAL 27-Squamous cell carcinoma
0.0

adenocarcinoma

of tongue

[0485]

43

TABLE IE

Panel 4D

Rel. Exp. (%) Ag2349,

Rel. Exp. (%) Ag2349,

Tissue Name
Run 160657277
Tissue Name
Run 160657277

Secondary Th1 act
0.0
HUVEC IL-1beta
0.0

Secondary Th2 act
0.0
HUVEC IFN gamma
0.0

Secondary Tr1 act
0.0
HUVEC TNF alpha + IFN
0.0

gamma

Secondary Th1 rest
0.0
HUVEC TNF alpha + IL4
0.0

Secondary Th2 rest
0.0
HUVEC IL-11
0.0

Secondary Tr1 rest
0.0
Lung Microvascular EC none
0.0

Primary Th1 act
0.0
Lung Microvascular EC
0.0

TNF alpha + IL-1beta

Primary Th2 act
0.0
Microvascular Dermal EC none
0.0

Primary Tr1 act
0.0
Microsvasular Dermal EC
0.0

TNF alpha + IL-1beta

Primary Th1 rest
0.0
Bronchial epithelium TNF alpha +
0.0

IL1beta

Primary Th2 rest
0.0
Small airway epithelium none
0.0

Primary Tr1 rest
0.0
Small airway epithelium
0.0

TNF alpha + IL-1beta

CD45RA CD4 lymphocyte
0.0
Coronery artery SMC rest
0.0

act

CD45RO CD4 lymphocyte
0.0
Coronery artery SMC TNF alpha +
0.0

act

IL-1beta

CD8 lymphocyte act
0.0
Astrocytes rest
0.0

Secondary CD8
0.0
Astrocytes TNF alpha + IL-1beta
0.0

lymphocyte rest

Secondary CD8
0.0
KU-812 (Basophil) rest
0.0

lymphocyte act

CD4 lymphocyte none
0.0
KU-812 (Basophil)
0.0

PMA/ionomycin

2ry Th1/Th2/Tr1 anti-
0.0
CCD1106 (Keratinocytes) none
0.0

CD95 CH11

LAK cells rest
0.0
CCD1106 (Keratinocytes)
0.0

TNF alpha + IL-1beta

LAK cells IL-2
0.0
Liver cirrhosis
100.0

LAK cells IL-2 + IL-12
0.0
Lupus kidney
0.0

LAK cells IL-2 + IFN
0.0
NCI-H292 none
0.0

gamma

LAK cells IL-2 + IL-18
0.0
NCI-H292 IL-4
0.0

LAK cells
0.0
NCI-H292 IL-9
0.0

PMA/ionomycin

NK Cells IL-2 rest
0.0
NCI-H292 IL-13
25.2

Two Way MLR 3 day
0.0
NCI-H292 IFN gamma
0.0

Two Way MLR 5 day
0.0
HPAEC none
0.0

Two Way MLR 7 day
0.0
HPAEC TNF alpha + IL-1beta
0.0

PBMC rest
0.0
Lung fibroblast none
0.0

PBMC PWM
0.0
Lung fibroblast TNF alpha + IL-
0.0

1beta

PBMC PHA-L
0.0
Lung fibroblast IL-4
0.0

Ramos (B cell) none
0.0
Lung fibroblast IL-9
0.0

Ramos (B cell) ionomycin
0.0
Lung fibroblast IL-13
0.0

B lymphocytes PWM
0.0
Lung fibroblast IFN gamma
29.1

B lymphocytes CD40L
0.0
Dermal fibroblast CCD1070 rest
0.0

and IL-4

EOL-1 dbcAMP
0.0
Dermal fibroblast CCD1070
0.0

TNF alpha

EOL-1 dbcAMP
0.0
Dermal fibroblast CCD1070 IL-
0.0

PMA/ionomycin

1beta

Dendritic cells none
0.0
Dermal fibroblast IFN gamma
0.0

Dendritic cells LPS
36.3
Dermal fibroblast IL-4
0.0

Dendritic cells anti-CD40
0.0
IBD Colitis 2
0.0

Monocytes rest
0.0
IBD Crohn's
0.0

Monocytes LPS
0.0
Colon
0.0

Macrophages rest
0.0
Lung
0.0

Macrophages LPS
0.0
Thymus
0.0

HUVEC none
0.0
Kidney
0.0

HUVEC starved
0.0

[0486] CNS_neurodegeneration_v1.0 Summary: Ag2349 Expression of the CG55978-01 gene is low/undetectable (CTs>35) across all of the samples on this panel.

[0487] Panel 1.3D Summary: Ag2349 Low but significant expression of the CG55978-01 is limited to three lung cancer cell lines, in two experiments with the same probe and primer set. Therefore, expression of this gene may be used to distinguish lung cancer cell lines from the other samples on this panel. Furthermore, therapeutic modulation of the activity of this GPCR could be beneficial in the treatment of lung cancer.

[0488] Panel 2D Summary: Ag2349 The CG55978-01 gene is expressed at significant levels in samples from normal prostate and prostate tumors. This observation suggests that expression of this gene may be used to distinguish prostate for other samples. Expression of this gene is also higher in a single lung cancer sample when compared to the matched adjacent tissue. Therefore, therapeutic modulation of the activity of the GPCR encoded by this gene could be beneficial in the treatment of lung cancer.

[0489] Panel 3D Summary: Ag2349 The CG55978-01 gene is most highly expressed in a small cell lung cancer cell line (CT=29). It is also expressed at low levels in two other lung cancer cell lines. This result is consistent with what is observed in Panel 1.3D. Therefore, expression of this gene may be used to distinguish lung cancer cell lines from the other samples on this panel. Furthermore, therapeutic modulation of the activity of this GPCR could be beneficial in the treatment of lung cancer

[0490] Panel 4D Summary: Ag2349 Low but significant expression of the CG55978-01 gene is limited to a liver cirrhosis sample (CT=34.7). Furthermore, expression of this gene is not detected in normal liver in Panel 1.3D, suggesting that its expression is unique to liver cirrhosis. This gene encodes a putative GPCR; therefore, antibodies or small molecule therapeutics could reduce or inhibit fibrosis that occurs in liver cirrhosis. In addition, antibodies to this putative GPCR could also be used for the diagnosis of liver cirrhosis.

[0491] J. CG56968-02/GMAC011711_E: Olfactory Receptor

[0492] Expression of gene CG56968-02 was assessed using the primer-probe set Ag2348, described in Table JA. Results of the RTQ-PCR runs are shown in Tables JB and JC.

44TABLE JAProbe Name Ag2348StartSEQ IDPrimersSequencesLengthPositionNO:Forward5′-tctctgcagtgctcctcttcta-3′22776204ProbeTET-5′-tccctatgatcctcctggcactgatt-3′26800205Reverse5′-tatgggtatgctgagtgattgg-3′22841206

[0493]

45

TABLE JB

Panel 1.3D

Rel. Exp. (%) Ag2348, Run

Rel. Exp. (%) Ag2348, Run

Tissue Name
165974844
Tissue Name
165974844

Liver adenocarcinoma
0.0
Kidney (fetal)
3.2

Pancreas
0.0
Renal ca. 786-0
0.0

Pancreatic ca. CAPAN 2
0.0
Renal ca. A498
0.0

Adrenal gland
0.0
Renal ca. RXF 393
0.0

Thyroid
0.0
Renal ca. ACHN
0.0

Salivary gland
0.0
Renal ca. UO-31
32.1

Pituitary gland
0.0
Renal ca.TK-10
0.0

Brain (fetal)
0.0
Liver
0.0

Brain (whole)
0.0
Liver (fetal)
0.0

Brain (amygdala)
0.0
Liver ca. (hepatoblast)
0.0

HepG2

Brain (cerebellum)
0.0
Lung
0.0

Brain (hippocampus)
0.0
Lung (fetal)
0.0

Brain (substantia nigra)
0.0
Lung ca. (small cell) LX-1
0.0

Brain (thalamus)
0.0
Lung ca. (small cell)
30.1

NCI-H69

Cerebral Cortex
0.0
Lung ca. (s.cell var.)
90.1

SHP-77

Spinal cord
0.0
Lung ca. (large cell)NCI-
0.0

H460

glio/astro U87-MG
0.0
Lung ca. (non-sm. cell)
0.0

A549

glio/astro U-118-MG
0.0
Lung ca. (non-s.cell)
0.0

NCI-H23

astrocytoma SW1783
0.0
Lung ca. (non-s.cell)
0.0

HOP-62

neuro*; met SK-N-AS
0.0
Lung ca. (non-s.cl) NCI-
0.0

H522

astrocytoma SF-539
0.0
Lung ca. (squam.) SW
0.0

900

astrocytoma SNB-75
0.0
Lung ca. (squam.) NCI-
100.0

H596

glioma SNB-19
0.0
Mammary gland
0.0

glioma U251
0.0
Breast ca.* (pl.ef) MCF-7
0.0

glioma SF-295
0.0
Breast ca.* (pl.ef) MDA-
0.0

MB-231

Heart (Fetal)
0.0
Breast ca.* (pl. ef) T47D
0.0

Heart
0.0
Breast ca. BT-549
0.0

Skeletal muscle (Fetal)
0.0
Breast ca. MDA-N
0.0

Skeletal muscle
0.0
Ovary
0.0

Bone marrow
0.0
Ovarian ca. OVCAR-3
0.0

Thymus
0.0
Ovarian ca. OVCAR-4
0.0

Spleen
0.0
Ovarian ca. OVCAR-5
0.0

Lymph node
0.0
Ovarian ca. OVCAR-8
0.0

Colorectal
18.6
Ovarian ca. IGROV-1
0.0

Stomach
0.0
Ovarian ca. (ascites) SK-
0.0

OV-3

Small intestine
0.0
Uterus
0.0

Colon ca. SW480
0.0
Placenta
0.0

Colon ca.* SW620
0.0
Prostate
0.0

(SW480 met)

Colon ca. HT29
0.0
Prostate ca.* (bone met)
0.0

PC-3

Colon ca. HCT-116
0.0
Testis
0.0

Colon ca. CaCo-2
0.0
Melanoma Hs688(A).T
0.0

CC Well to Mod Diff
0.0
Melanoma* (met)
0.0

(ODO3866)

Hs688(B).T

Colon ca. HCC-2998
0.0
Melanoma UACC-62
0.0

Gastric ca. (liver met)
0.0
Melanoma M14
0.0

NCI-N87

Bladder
0.0
Melanoma LOX IMVI
0.0

Trachea
0.0
Melanoma* (met) SK-
0.0

MEL-5

Kidney
0.0
Adipose
0.0

[0494]

46

TABLE JC

Panel 4D

Ref. Exp. (%) Ag2348,

Rel. Exp. (%) Ag2348,

Tissue Name
Run 164023294
Tissue Name
Run 164023294

Secondary Th1 act
0.0
HUVEC IL-1beta
0.0

Secondary Th2 act
0.0
HUVEC IFN gamma
0.0

Secondary Tr1 act
0.0
HUVEC TNF alpha + IFN
0.0

gamma

Secondary Th1 rest
0.0
HUVEC TNF alpha + IL4
0.0

Secondary Th2 rest
0.0
HUVEC IL-11
0.0

Secondary Tr1 rest
0.0
Lung Microvascular EC none
0.0

Primary Th1 act
23.5
Lung Microvascular EC
0.0

TNF alpha + IL-1beta

Primary Th2 act
0.0
Microvascular Dermal EC none
0.0

Primary Tr1 act
0.0
Microsvasular Dermal EC
0.0

TNF alpha + IL-1beta

Primary Th1 rest
0.0
Bronchial epithelium TNF alpha +
0.0

IL1beta

Primary Th2 rest
0.0
Small airway epithelium none
0.0

Primary Tr1 rest
0.0
Small airway epithelium
0.0

TNF alpha + IL-1beta

CD45RA CD4 lymphocyte
0.0
Coronery artery SMC rest
0.0

act

CD45RO CD4 lymphocyte
0.0
Coronery artery SMC TNF alpha +
0.0

act

IL-1beta

CD8 lymphocyte act
0.0
Astrocytes rest
0.0

Secondary CD8
0.0
Astrocytes TNF alpha + IL-1beta
0.0

lymphocyte rest

Secondary CD8
0.0
KU-812 (Basophil) rest
0.0

lymphocyte act

CD4 lymphocyte none
0.0
KU-812 (Basophil)
0.0

PMA/ionomycin

2ry Th1/Th2/Tr1_anti-
0.0
CCD1106 (Keratinocytes) none
0.0

CD95 CH11

LAK cells rest
0.0
CCD1106 (Keratinocytes)
0.0

TNF alpha + IL-1beta

LAK cells IL-2
0.0
Liver cirrhosis
100.0

LAK cells IL-2 + IL-12
0.0
Lupus kidney
0.0

LAK cells IL-2 + IFN
0.0
NCI-H292 none
0.0

gamma

LAK cells IL-2 + IL-18
0.0
NCI-H292 IL-4
0.0

LAK cells
0.0
NCI-H292 IL-9
0.0

PMA/ionomycin

NK Cells IL-2 rest
0.0
NCI-H292 IL-13
0.0

Two Way MLR 3 day
0.0
NCI-H292 IFN gamma
0.0

Two Way MLR 5 day
0.0
HPAEC none
0.0

Two Way MLR 7 day
0.0
HPAEC TNF alpha + IL-1beta
0.0

PBMC rest
0.0
Lung fibroblast none
0.0

PBMC PWM
0.0
Lung fibroblast TNF alpha + IL-
0.0

1beta

PBMC PHA-L
0.0
Lung fibroblast IL-4
5.3

Ramos (B cell) none
0.0
Lung fibroblast IL-9
0.0

Ramos (B cell) ionomycin
0.0
Lung fibroblast IL-13
0.0

B lymphocytes PWM
0.0
Lung fibroblast IFN gamma
0.0

B lymphocytes CD40L
0.0
Dermal fibroblast CCD1070 rest
0.0

and IL-4

EOL-1 dbcAMP
0.0
Dermal fibroblast CCD1070
0.0

TNF alpha

EOL-1 dbcAMP
0.0
Dermal fibroblast CCD1070 IL-
0.0

PMA/ionomycin

1beta

Dendritic cells none
0.0
Dermal fibroblast IFN gamma
0.0

Dendritic cells LPS
0.0
Dermal fibroblast IL-4
0.0

Dendritic cells anti-CD40
0.0
IBD Colitis 2
0.0

Monocytes rest
0.0
IBD Crohn's
0.0

Monocytes LPS
0.0
Colon
0.0

Macrophages rest
0.0
Lung
13.8

Macrophages LPS
0.0
Thymus
0.0

HUVEC none
0.0
Kidney
0.0

HUVEC starved
0.0

[0495] Panel 1.3D Summary: Ag2348 Low but significant expression of the CG56968-02 gene is limited to two lung cancer cell lines (CTs=34.1). Therefore, expression of this gene may be used to distinguish lung cancer cell lines from the other samples on this panel. Furthermore, therapeutic modulation of the activity of this GPCR could be beneficial in the treatment of lung cancer.

[0496] Panel 2.2 Summary: Ag2348 Expression of the CG56968-02 gene is low/undetectable (CTs>35) across all of the samples on this panel. (Data not shown.)

[0497] Panel 4D Summary: Ag2348 Low but significant expression of the CG56968-02 gene is limited to a liver cirrhosis sample (CT=33). Furthermore, expression of this gene is not detected in normal liver in Panel 1.3D, suggesting that its expression is unique to liver cirrhosis. This gene encodes a putative GPCR; therefore, antibodies or small molecule therapeutics could reduce or inhibit fibrosis that occurs in liver cirrhosis. In addition, antibodies to this putative GPCR could also be used for the diagnosis of liver cirrhosis.

[0498] K CG50197-03/GMAC011711_D: Olfactory Receptor

[0499] Expression of gene CG50197-03 was assessed using the primer-probe sets Ag2347 and Ag2482, described in Tables KA and KB. Results of the RTQ-PCR runs are shown in Tables KC, KD, KE, KF, and KG.

47TABLE KAProbe Name Ag2347StartSEQ IDPrimersSequencesLengthPositionNO:Forward5′-gtctccatggctggatctctat-3′2273207ProbeTET-5′-tcccttctgcttcatctacctgacag-3′-TAMRA2695208Reverse5′-tacaaatgacgtggagaatggt-3′22138209

[0500]

48

TABLE KB

Probe Name Ag2482

Start
SEQ ID

Primers
Sequences
Length
Position
NO:

Forward
5′-gatgccaatctctctggtatca-3′
22
269
210

Probe
TET-5′-aaccagaaaatgcccagcacagtg-3′-TAMRA
24
245
211

Reverse
5′-ctgtcacagacttaggcctttg-3′
22
208
212

[0501]

49

TABLE KC

Panel 1.3D

Rel. Exp. (%)
Rel. Exp. (%)

Rel. Exp. (%)
Rel. Exp. (%)

Ag2482, Run
Ag2482, Run

Ag2482, Run
Ag2482, Run

Tissue Name
163724418
165639464
Tissue Name
163724418
165639464

Liver
0.0
0.0
Kidney (fetal)
2.4
0.0

adenocarcinoma

Pancreas
0.0
0.0
Renal ca. 786-0
0.0
0.0

Pancreatic ca.
0.0
0.0
Renal ca. A498
0.0
0.0

CAPAN 2

Adrenal gland
0.0
0.0
Renal ca. RXF
0.0
0.0

393

Thyroid
0.0
0.0
Renal ca. ACHN
0.0
0.0

Salivary gland
0.0
0.0
Renal ca. UO-31
6.4
0.0

Pituitary gland
0.0
0.0
Renal ca.TK-10
0.0
0.0

Brain (fetal)
0.0
0.0
Liver
0.0
0.0

Brain (whole)
0.0
0.0
Liver (fetal)
0.0
0.0

Brain (amygdala)
0.0
0.0
Liver ca.
0.0
0.0

(hepatoblast)

HepG2

Brain (Cerebellum)
0.0
0.0
Lung
0.0
0.0

Brain
0.0
0.0
Lung (fetal)
0.0
0.0

(hippocampus)

Brain (substantia
0.0
0.0
Lung ca. (small
0.0
0.0

nigra)

cell) LX-1

Brain (thalamus)
0.0
0.0
Lung ca. (small
16.0
2.2

cell) NCI-H69

Cerebral Cortex
0.0
0.0
Lung ca. (s.cell
100.0
100.0

var.) SHP-77

Spinal cord
0.0
0.0
Lung ca. (large
0.0
0.0

cell)NCI-H460

glio/astro U87-MG
0.0
0.0
Lung ca. (non-
0.0
0.0

sm. cell) A549

glio/astro U-118-
0.0
0.0
Lung ca. (non-
0.0
0.0

MG

s.cell) NCI-H23

astrocytoma
0.0
0.0
Lung ca. (non-
0.0
0.0

SW1783

s.cell) HOP-62

neuro*; met SK-N-
0.0
2.4
Lung ca. (non-
0.0
0.0

AS

s.cl) NCI-H522

astrocytoma SF-539
0.0
3.8
Lung ca.
0.0
0.0

(squam.) SW 900

astrocytoma SNB-
0.0
0.0
Lung ca.
0.0
11.5

75

(squam.) NCI-

H596

glioma SNB-19
0.0
0.0
Mammary gland
0.0
0.0

glioma U251
0.0
0.0
Breast ca.*
0.0
0.0

(pl.ef) MCF-7

glioma SF-295
0.0
0.0
Breast ca.*
0.0
0.0

(pl.ef) MDA-

MB-231

Heart (Fetal)
0.0
0.0
Breast ca.* (pl.
0.0
0.0

ef) T47D

Heart
0.0
0.0
Breast ca. BT-
0.0
3.7

549

Skeletal muscle
0.0
0.0
Breast ca. MDA-N
0.0
0.0

(Fetal)

Skeletal muscle
0.0
0.0
Ovary
0.0
0.0

Bone marrow
0.0
0.0
Ovarian ca.
0.0
0.0

OVCAR-3

Thymus
0.0
0.0
Ovarian ca.
0.0
0.0

OVCAR-4

Spleen
0.0
0.0
Ovarian ca.
0.0
0.0

OVCAR-5

Lymph node
0.0
0.0
Ovarian ca.
0.0
0.0

OVCAR-8

Colorectal
2.6
0.0
Ovarian ca.
0.0
0.0

IGROV-1

Stomach
0.0
0.0
Ovarian ca.
0.0
0.0

(ascites) SK-OV-3

Small intestine
0.0
0.0
Uterus
0.0
0.0

Colon ca. SW480
1.9
0.0
Placenta
0.0
0.0

Colon ca.* SW620
0.0
0.0
Prostate
5.7
0.0

(SW480 met)

Colon ca. HT29
0.0
0.0
Prostate ca.*
0.0
0.0

(bone met) PC-3

Colon ca. HCT-116
0.0
0.0
Testis
2.3
3.4

Colon ca. CaCo-2
0.0
0.0
Melanoma
0.0
0.0

Hs688(A).T

CC Well to Mod
0.0
4.1
Melanoma*
0.0
0.0

Diff (ODO3866)

(met)

Hs688(B).T

Colon ca. HCC-
0.0
0.0
Melanoma
0.0
0.0

2998

UACC-62

Gastric ca. (liver
0.0
0.0
Melanoma M14
0.0
0.0

met) NCI-N87

Bladder
4.9
0.0
Melanoma LOX
0.0
0.0

IMVI

Trachea
2.0
0.0
Melanoma*
0.0
0.0

(met) SK-MEL-5

Kidney
0.0
0.0
Adipose
0.0
0.0

[0502]

50

TABLE KD

Panel 2.2

Rel. Exp. (%) Ag2347,

Rel. Exp. (%) Ag2347,

Tissue Name
Run 174294963
Tissue Name
Run 174294963

Normal Colon
0.0
Kidney Margin (OD04348)
0.0

Colon cancer (OD06064)
0.0
Kidney malignant cancer
0.0

(OD06204B)

Colon Margin (OD06064)
0.0
Kidney normal adjacent
0.0

tissue (OD06204E)

Colon cancer (OD06159)
0.0
Kidney Cancer (OD04450-
0.0

01)

Colon Margin (OD06159)
0.0
Kidney Margin (OD04450-
0.0

03)

Colon cancer (OD06297-04)
0.0
Kidney Cancer 8120613
0.0

Colon Margin (OD06297-
0.0
Kidney Margin 8120614
0.0

015)

CC Gr.2 ascend colon
100.0
Kidney Cancer 9010320
0.0

(ODO3921)

CC Margin (ODO3921)
0.0
Kidney Margin 9010321
0.0

Colon cancer metastasis
0.0
Kidney Cancer 8120607
0.0

(OD06104)

Lung Margin (OD06104)
0.0
Kidney Margin 8120608
0.0

Colon mets to lung
0.0
Normal Uterus
0.0

(OD04451-01)

Lung Margin (OD04451-02)
0.0
Uterine Cancer 064011
0.0

Normal Prostate
0.0
Normal Thyroid
0.0

Prostate Cancer (OD04410)
0.5
Thyroid Cancer
0.0

Prostate Margin (OD04410)
0.9
Thyroid Cancer A302152
0.0

Normal Ovary
0.0
Thyroid Margin A302153
0.8

Ovarian cancer (OD06283-
0.0
Normal Breast
0.4

03)

Ovarian Margin (OD06283-
0.0
Breast Cancer
0.0

07)

Ovarian Cancer
0.6
Breast Cancer
0.0

Ovarian cancer (OD06145)
0.0
Breast Cancer (OD04590-
0.0

01)

Ovarian Margin (OD06145)
0.0
Breast Cancer Mets
0.0

(OD04590-03)

Ovarian cancer (OD06455-
0.0
Breast Cancer Metastasis
0.0

03)

Ovarian Margin (OD06455-
0.0
Breast Cancer
0.0

07)

Normal Lung
0.0
Breast Cancer 9100266
0.0

Invasive poor diff. lung
0.0
Breast Margin 9100265
0.0

adeno (ODO4945-01

Lung Margin (ODO4945-03)
0.0
Breast Cancer A209073
0.0

Lung Malignant Cancer
0.0
Breast Margin A2090734
0.0

(OD03126)

Lung Margin (OD03126)
0.0
Breast cancer (OD06083)
0.0

Lung Cancer (OD05014A)
0.0
Breast cancer node
0.0

metastasis (OD06083)

Lung Margin (OD05014B)
0.0
Normal Liver
0.0

Lung cancer (OD06081)
0.0
Liver Cancer 1026
0.0

Lung Margin (OD06081)
0.0
Liver Cancer 1025
0.0

Lung Cancer (OD04237-01)
0.0
Liver Cancer 6004-T
0.0

Lung Margin (OD04237-02)
0.0
Liver Tissue 6004-N
0.0

Ocular Mel Met to Liver
0.0
Liver Cancer 6005-T
0.0

(ODO4310)

Liver Margin (ODO4310)
0.0
Liver Tissue 6005-N
0.0

Melanoma Metastasis
0.0
Liver Cancer
0.0

Lung Margin (OD04321)
0.0
Normal Bladder
0.0

Normal Kidney
0.0
Bladder Cancer
0.0

Kidney Ca, Nuclear grade 2
0.0
Bladder Cancer
0.0

(OD04338)

Kidney Margin (OD04338)
0.0
Normal Stomach
0.0

Kidney Ca Nuclear grade 1/2
0.0
Gastric Cancer 9060397
0.0

(OD04339)

Kidney Margin (OD04339)
0.0
Stomach Margin 9060396
0.0

Kidney Ca, Clear cell type
0.0
Gastric Cancer 9060395
0.0

(OD04340)

Kidney Margin (OD04340)
0.0
Stomach Margin 9060394
0.0

Kidney Ca, Nuclear grade 3
0.0
Gastric Cancer 064005
0.0

(OD04348)

[0503]

51

TABLE KE

Panel 2D

Rel. Exp.

Rel. Exp.

(%) Ag2482,

(%) Ag2482,

Tissue Name
Run 162558432
Tissue Name
Run 162558432

Normal Colon
8.4
Kidney Margin 8120608
0.0

CC Well to Mod Diff
7.4
Kidney Cancer 8120613
0.0

(ODO3866)

CC Margin (ODO3866)
6.7
Kidney Margin 8120614
0.0

CC Gr.2 rectosigmoid
2.5
Kidney Cancer 9010320
0.0

(ODO3868)

CC Margin (ODO3868)
0.0
Kidney Margin 9010321
0.0

CC Mod Diff (ODO3920)
0.0
Normal Uterus
0.0

CC Margin (ODO3920)
0.0
Uterine Cancer 064011
0.0

CC Gr.2 ascend colon
0.0
Normal Thyroid
0.0

(ODO3921)

CC Margin (ODO3921)
4.6
Thyroid Cancer
0.0

CC from Partial Hepatectomy
0.0
Thyroid Cancer A302152
0.0

(ODO4309) Mets

Liver Margin (ODO4309)
0.0
Thyroid Margin A302153
0.0

Colon mets to lung (OD04451-
0.0
Normal Breast
0.0

01)

Lung Margin (OD04451-02)
0.0
Breast Cancer
0.0

Normal Prostate 6546-1
32.8
Breast Cancer (OD04590-
0.0

01)

Prostate Cancer (OD04410)
100.0
Breast Cancer Mets
0.0

(OD04590-03)

Prostate Margin (OD04410)
44.8
Breast Cancer Metastasis
7.2

Prostate Cancer (OD04720-01)
9.7
Breast Cancer
0.0

Prostate Margin (OD04720-02)
62.0
Breast Cancer
0.0

Normal Lung
0.0
Breast Cancer 9100266
0.0

Lung Met to Muscle
12.2
Breast Margin 9100265
0.0

(ODO4286)

Muscle Margin (ODO4286)
0.0
Breast Cancer A209073
0.0

Lung Malignant Cancer
25.5
Breast Margin A2090734
0.0

(OD03126)

Lung Margin (OD03126)
0.0
Normal Liver
0.0

Lung Cancer (OD04404)
0.0
Liver Cancer
0.0

Lung Margin (OD04404)
0.0
Liver Cancer 1025
0.0

Lung Cancer (OD04565)
0.0
Liver Cancer 1026
0.0

Lung Margin (OD04565)
0.0
Liver Cancer 6004-T
2.4

Lung Cancer (OD04237-01)
0.0
Liver Tissue 6004-N
0.0

Lung Margin (OD04237-02)
0.0
Liver Cancer 6005-T
0.0

Ocular Mel Met to Liver
0.0
Liver Tissue 6005-N
0.0

(ODO4310)

Liver Margin (ODO4310)
0.0
Normal Bladder
2.1

Melanoma Metastasis
0.0
Bladder Cancer
0.0

Lung Margin (OD04321)
0.0
Bladder Cancer
18.7

Normal Kidney
0.0
Bladder Cancer
0.0

(OD04718-01)

Kidney Ca, Nuclear grade 2
0.0
Bladder Normal Adjacent
0.0

(OD04338)

(OD04718-03)

Kidney Margin (OD04338)
3.3
Normal Ovary
0.0

Kidney Ca Nuclear grade 1/2
0.0
Ovarian Cancer
0.0

(OD04339)

Kidney Margin (OD04339)
0.0
Ovarian Cancer
0.0

(OD04768-07)

Kidney Ca, Clear cell type
5.1
Ovary Margin (OD04768-
0.0

(OD04340)

08)

Kidney Margin (OD04340)
0.0
Normal Stomach
0.0

Kidney Ca, Nuclear grade 3
0.0
Gastric Cancer 9060358
0.0

(OD04348)

Kidney Margin (OD04348)
0.0
Stomach Margin 9060359
0.0

Kidney Cancer (OD04622-01)
0.0
Gastric Cancer 9060395
0.0

Kidney Margin (OD04622-03)
0.0
Stomach Margin 9060394
0.0

Kidney Cancer (OD04450-01)
0.0
Gastric Cancer 9060397
2.6

Kidney Margin (OD04450-03)
0.0
Stomach Margin 9060396
0.0

Kidney Cancer 8120607
0.0
Gastric Cancer 064005
0.0

[0504]

52

TABLE KF

Panel 3D

Rel. Exp. (%)

Rel. Exp. (%)

Ag2482, Run

Ag2482, Run

Tissue Name
164632281
Tissue Name
164632281

Daoy-Medulloblastoma
0.0
Ca Ski-Cervical epidermoid
0.0

carcinoma (metastasis)

TE671-Medulloblastoma
0.0
ES-2-Ovarian clear cell carcinoma
0.0

D283 Med-Medulloblastoma
0.0
Ramos-Stimulated with
0.0

PMA/ionomycin 6 h

PFSK-1-Primitive
0.0
Ramos-Stimulated with
0.0

Neuroectodermal

PMA/ionomycin 14 h

XF-498-CNS
0.0
MEG-01-Chronic myelogenous
0.0

leukemia (megokaryoblast)

SNB-78-Glioma
0.0
Raji-Burkitt's lymphoma
0.0

SF-268-Glioblastoma
0.0
Daudi-Burkitt's lymphoma
0.0

T98G-Glioblastoma
0.0
U266-B-cell plasmacytoma
0.0

SK-N-SH-Neuroblastoma
0.0
CA46-Burkitt's lymphoma
0.0

(metastasis)

SF-295-Glioblastoma
0.0
RL-non-Hodgkin's B-cell
0.0

lymphoma

Cerebellum
0.0
JM1-pre-B-cell lymphoma
0.0

Cerebellum
0.0
Jurkat-T cell leukemia
0.0

NCI-H292-Mucoepidermoid
0.0
TF-1-Erythroleukemia
0.0

lung carcinoma

DMS-114-Small cell lung
0.0
HUT 78-T-cell lymphoma
0.0

cancer

DMS-79-Small cell lung
100.0
U937-Histiocytic lymphoma
0.0

cancer

NCI-H146-Small cell lung
12.7
KU-812-Myelogenous leukemia
0.0

cancer

NCI-H526-Small cell lung
4.4
769-P-Clear cell renal carcinoma
0.0

cancer

NCI-N417-Small cell lung
0.0
Caki-2-Clear cell renal carcinoma
0.0

cancer

NCI-H82-Small cell lung
0.0
SW 839-Clear cell renal carcinoma
0.0

cancer

NCI-H157-Squamous cell
0.0
G401-Wilms' tumor
0.0

lung cancer (metastasis)

NCI-H1155-Large cell lung
0.0
Hs766T-Pancreatic carcinoma (LN
0.0

cancer

metastasis)

NCI-H1299-Large cell lung
0.0
CAPAN-1-Pancreatic
0.0

cancer

adenocarcinoma (liver metastasis)

NCI-H727-Lung carcinoid
0.0
SU86.86-Pancreatic carcinoma
0.0

(liver metastasis)

NCI-UMC-11-Lung
8.8
BxPC-3-Pancreatic
0.0

carcinoid

adenocarcinoma

LX-1-Small cell lung cancer
0.0
HPAC-Pancreatic adenocarcinoma
0.0

Colo-205-Colon cancer
0.0
MIA PaCa-2-Pancreatic carcinoma
0.0

KM12-Colon cancer
0.0
CFPAC-1-Pancreatic ductal
0.0

adenocarcinoma

KM20L2-Colon cancer
0.0
PANC-1-Pancreatic epithelioid
0.0

ductal carcinoma

NCI-H716-Colon cancer
0.0
T24-Bladder carcinma (transitional
0.0

cell)

SW-48-Colon
0.0
5637-Bladder carcinoma
0.0

adenocarcinoma

SW1116-Colon
0.0
HT-1197-Bladder carcinoma
0.0

adenocarcinoma

LS 174T-Colon
0.0
UM-UC-3-Bladder carcinoma
0.0

adenocarcinoma

(transitional cell)

SW-948-Colon
0.0
A204-Rhabdomyosarcoma
0.0

adenocarcinoma

SW-480-Colon
0.0
HT-1080-Fibrosarcoma
0.0

adenocarcinoma

NCI-SNU-5-Gastric
0.0
MG-63-Osteosarcoma
8.5

carcinoma

KATO III-Gastric carcinoma
0.0
SK-LMS-1-Leiomyosarcoma
0.0

(vulva)

NCI-SNU-16-Gastric
0.0
SJRH30-Rhabdomyosarcoma (met
0.0

carcinoma

to bone marrow)

NCI-SNU-1-Gastric
0.0
A431-Epidermoid carcinoma
0.0

carcinoma

RF-1-Gastric
0.0
WM266-4-Melanoma
0.0

adenocarcinoma

RF-48-Gastric
0.0
DU 145-Prostate carcinoma (brain
0.0

adenocarcinoma

metastasis)

MKN-45-Gastric carcinoma
0.0
MDA-MB-468-Breast
0.0

adenocarcinoma

NCI-N87-Gastric carcinoma
0.0
SCC-4-Squamous cell carcinoma
0.0

of tongue

OVCAR-5-Ovarian
0.0
SCC-9-Squamous cell carcinoma
0.0

carcinoma

of tongue

RL95-2-Uterine carcinoma
0.0
SCC-15-Squamous cell carcinoma
0.0

of tongue

HelaS3-Cervical
0.0
CAL 27-Squamous cell carcinoma
0.0

adenocarcinoma

of tongue

[0505]

53

TABLE KG

Panel 4.1D

Rel. Exp.

Rel. Exp.

(%) Ag2347,

(%) Ag2347,

Tissue Name
Run 224781589
Tissue Name
Run 224781589

Secondary Th1 act
0.0
HUVEC IL-1beta
0.0

Secondary Th2 act
0.0
HUVEC IFN gamma
0.0

Secondary Tr1 act
0.0
HUVEC TNF alpha + IFN
0.0

gamma

Secondary Th1 rest
0.0
HUVEC TNF alpha + IL4
0.0

Secondary Th2 rest
0.0
HUVEC IL-11
0.0

Secondary Tr1 rest
0.0
Lung Microvascular EC none
0.0

Primary Th1 act
0.0
Lung Microvascular EC
0.7

TNF alpha + IL-1beta

Primary Th2 act
0.0
Microvascular Dermal EC none
0.0

Primary Tr1 act
0.0
Microsvasular Dermal EC
0.0

TNF alpha + IL-1beta

Primary Th1 rest
0.0
Bronchial epithelium
0.0

TNF alpha + IL1beta

Primary Th2 rest
0.0
Small airway epithelium none
0.0

Primary Tr1 rest
0.0
Small airway epithelium
0.0

TNF alpha + IL-1beta

CD45RA CD4 lymphocyte
0.0
Coronery artery SMC rest
0.0

act

CD45RO CD4 lymphocyte
0.0
Coronery artery SMC TNF
0.0

act

alpha + IL-1beta

CD8 lymphocyte act
0.0
Astrocytes rest
0.0

Secondary CD8
0.0
Astrocytes TNF alpha + IL-1beta
0.0

lymphocyte rest

Secondary CD8
0.0
KU-812 (Basophil) rest
0.0

lymphocyte act

CD4 lymphocyte none
0.0
KU-812 (Basophil)
0.0

PMA/ionomycin

2ry Th1/Th2/Tr1_anti-
0.0
CCD1106 (Keratinocytes) none
0.0

CD95 CH11

LAK cells rest
0.0
CCD1106 (Keratinocytes)
0.0

TNF alpha + IL-1beta

LAK cells IL-2
0.0
Liver cirrhosis
0.0

LAK cells IL-2 + IL-12
0.0
NCI-H292 none
0.0

LAK cells IL-2 + IFN
0.0
NCI-H292 IL-4
0.0

gamma

LAK cells IL-2 + IL-18
0.0
NCI-H292 IL-9
0.0

LAK cells
0.0
NCI-H292 IL-13
0.0

PMA/ionomycin

NK Cells IL-2 rest
0.0
NCI-H292 IFN gamma
0.0

Two Way MLR 3 day
0.0
HPAEC none
0.0

Two Way MLR 5 day
0.5
HPAEC TNF alpha + IL-1beta
0.0

Two Way MLR 7 day
0.0
Lung fibroblast none
0.0

PBMC rest
0.0
Lung fibroblast TNF alpha + IL-
0.0

1beta

PBMC PWM
0.0
Lung fibroblast IL-4
0.0

PBMC PHA-L
0.0
Lung fibroblast IL-9
0.0

Ramos (B cell) none
0.0
Lung fibroblast IL-13
0.0

Ramos (B cell) ionomycin
0.0
Lung fibroblast IFN gamma
0.0

B lymphocytes PWM
0.0
Dermal fibroblast CCD1070 rest
0.0

B lymphocytes CD40L
0.0
Dermal fibroblast CCD1070
0.0

and IL-4

TNF alpha

EOL-1 dbcAMP
0.0
Dermal fibroblast CCD1070 IL-
0.0

1beta

EOL-1 dbcAMP
0.0
Dermal fibroblast IFN gamma
0.0

PMA/ionomycin

Dendritic cells none
0.0
Dermal fibroblast IL-4
0.0

Dendritic cells LPS
0.0
Dermal Fibroblasts rest
0.0

Dendritic cells anti-CD40
0.0
Neutrophils TNFa + LPS
1.0

Monocytes rest
0.0
Neutrophils rest
1.4

Monocytes LPS
0.0
Colon
0.8

Macrophages rest
0.0
Lung
1.3

Macrophages LPS
0.0
Thymus
9.0

HUVEC none
0.0
Kidney
100.0

HUVEC starved
0.0

[0506] CNS_neurodegeneration_v1.0 Summary: Ag2347 Expression of the CG50197-03 gene in panel CNS_neurodegeneration_v1.0 is low/undetectable (CT values>35) in all samples (data not shown).

[0507] General_screening_panel_v1.5 Summary: Ag2347 Data from one experiment with this probe and primer set is not included because the amp plot suggests that there were experimental difficulties with this run. (Data not shown.)

[0508] Panel 1.3D Summary: Ag2482 Expression of the CG50197-03 gene in two independent runs is highest in a sample derived from a lung cancer cell line (SHP-77). Its expression in this panel is almost exclusive to this sample. Thus, the expression of the CG50197-03 gene could be used to distinguish samples derived from this cell line and other samples. Furthemore, therapeutic modulation of the expression or function of the protein encoded by the CG50197-03 gene, through the use of small molecule drugs or antibodies, may be useful in the treatment of lung cancer. Ag2347 Expression of the CG50197-03 gene in panel 1.3D is low/undetectable (CT values>35) in all samples (data not shown).

[0509] Panel 2.2 Summary: Ag2347 Expression of the CG50197-03 gene is limited to a sample derived from a colon cancer (CT=31.4). This result suggests that expression of the CG50197-03 gene could be used as a diagnostic marker for the presence of colon cancer. Furthermore, therapeutic modulation of the expression or function of the CG50197-03 gene product, through the use of small molecule drugs or antibodies, may be effective in the treatment of colon cancer. Ag2482 Expression of the CG50197-03 gene in Panel 2.2 is low/undetectable (CT values>35) in all samples (data not shown).

[0510] Panel 2D Summary: Ag2482 Expression of the CG50197-03 gene is highest in a sample derived from a prostate cancer and overall, its expression appears to be specific for prostate tissue. In addition, one sample derived from prostate cancer shows substantial over expression when compared to a matched sample derived from normal adjacent tissue. Thus, the expression of this gene could be used to distinguish prostate derived tissue from other tissues. Moreover, therapeutic modulation of the expression or function of the CG50197-03 gene or its protein product, through the use of small molecule drugs, antibodies or protein therapeutics, might be of use in the treatment of prostate cancer.

[0511] Panel 3D Summary: Ag2482 Significant expression of the CG50197-03 gene is limited to a sample derived from a lung cancer cell line. This preferential expression in lung cancer is also seen in the expression profiles from Panel 1.3D. This result suggests that expression of the CG50197-03 gene could be used to distinguish this cell line from other samples. Furthermore, therapeutic modulation of the gene or its protein product could potentially be useful in the treatment of lung cancer.

[0512] Panel 4.1D Summary: Ag2347 The CG50197-03 gene is only expressed at detectable levels in the kidney(CT=32). The putative GPCR encoded for by this gene could allow cells within the kidney to respond to specific microenvironmental signals (For example, ref. 1). Therefore, antibody or small molecule therapies designed with the protein encoded for by this gene could modulate kidney function and be important in the treatment of inflammatory or autoimmune diseases that affect the kidney, including lupus and glomerulonephritis (Mark et al., G protein modulation of recombinant P/Q-type calcium channels by regulators of G protein signalling proteins. J. Physiol. 528 Pt 1: 65-77, 2000).

[0513] Panel 4D Summary: Ag2482/Ag2347 Expression of the CG50197-03 gene in this panel is low/undetectable (CT values>35) in all samples (data not shown).

[0514] L. CG50217-02/GMAC011711_B: Olfactory Receptor

[0515] Expression of gene CG50217-02 was assessed using the primer-probe sets Ag2345 and Ag2494, described in Tables LA and LB. Results of the RTQ-PCR runs are shown in Tables LC, and LD.

54TABLE LAProbe Name Ag2345StartSEQ IDPrimersSequencesLengthPositionNO:Forward5′-gattttgagagctgtgcttcag-3′22672213ProbeTET-5′-cctcaaagcttttagcacacgtgcct-3′-TAMRA26714214Reverse5′-agccaagatgacacagatatgg-3′22741215

[0516]

55

TABLE LB

Probe Name Ag2494

Start
SEQ ID

Primers
Sequences
Length
Position
NO:

Forward
5′-ttccacaacatccttggataac-3′
22
919
216

Probe
TET-5′-ccccgatctgtttggttctaactcca-3′-TAMRA
26
888
217

Reverse
5′-tactgatacctcccatgctcaa-3′
22
854
218

[0517]

56

TABLE LC

Panel 1.3D

Rel. Exp. (%)
Rel. Exp. (%)

Rel. Exp. (%)
Rel. Exp. (%)

Ag2345, Run
Ag2494, Run

Ag2345, Run
Ag2494, Run

Tissue Name
165974938
165630640
Tissue Name
165974938
165630640

Liver
0.0
0.0
Kidney (fetal)
0.0
0.0

adenocarcinoma

Pancreas
0.0
0.0
Renal ca. 786-0
0.0
0.0

Pancreatic ca.
0.0
0.0
Renal ca. A498
1.4
0.0

CAPAN 2

Adrenal gland
0.0
7.4
Renal ca. RXF
0.0
0.0

393

Thyroid
0.0
0.0
Renal ca. ACHN
0.0
0.0

Salivary gland
0.0
0.0
Renal ca. UO-31
0.0
0.0

Pituitary gland
0.0
0.0
Renal ca. TK-10
0.0
0.0

Brain (fetal)
0.0
0.0
Liver
0.0
0.0

Brain (whole)
0.0
0.0
Liver (fetal)
0.0
0.0

Brain (amygdala)
0.0
0.0
Liver ca.
0.0
0.0

(hepatoblast)

HepG2

Brain (cerebellum)
0.0
0.0
Lung
0.0
0.0

Brain
0.0
0.0
Lung (fetal)
0.0
0.0

(hippocampus)

Brain (substantia
0.0
17.9
Lung ca. (small
0.0
0.0

nigra)

cell) LX-1

Brain (thalamus)
0.0
0.0
Lung ca. (small
48.3
0.0

cell) NCI-H69

Cerebral Cortex
0.0
0.0
Lung ca. (s.cell
100.0
0.0

var.) SHP-77

Spinal cord
0.0
21.3
Lung ca. (large
0.0
0.0

cell)NCI-H460

glio/astro U87-MG
0.0
0.0
Lung ca. (non-
0.0
0.0

sm. cell) A549

glio/astro U-118-
0.0
0.0
Lung ca. (non-
0.0
0.0

MG

s.cell) NCI-H23

astrocytoma
4.8
0.0
Lung ca. (non-
0.0
0.0

SW1783

s.cell) HOP-62

neuro*; met SK-N-
0.0
0.0
Lung ca. (non-
0.0
0.0

AS

s.cl) NCI-H522

astrocytoma SF-539
0.0
0.0
Lung ca.
0.0
15.6

(squam.) SW 900

astrocytoma SNB-
0.0
0.0
Lung ca.
63.7
0.0

75

(squam.) NCI-

H596

glioma SNB-19
2.3
0.0
Mammary gland
0.0
0.0

glioma U251
0.0
0.0
Breast ca.*
0.0
0.0

(pl.ef) MCF-7

glioma SF-295
0.0
0.0
Breast ca.*
0.0
15.5

(pl.ef) MDA-

MB-231

Heart (Fetal)
0.0
0.0
Breast ca.* (pl.
0.0
0.0

ef) T47D

Heart
0.0
0.0
Breast ca. BT-
0.0
0.0

549

Skeletal muscle
0.0
0.0
Breast ca. MDA-N
0.0
0.0

(Fetal)

Skeletal muscle
0.0
0.0
Ovary
0.0
0.0

Bone marrow
0.0
34.2
Ovarian ca.
0.0
0.0

OVCAR-3

Thymus
0.0
0.0
Ovarian ca.
0.0
0.0

OVCAR-4

Spleen
0.0
0.0
Ovarian ca.
0.0
0.0

OVCAR-5

Lymph node
0.0
4.5
Ovarian ca.
0.0
0.0

OVCAR-8

Colorectal
0.0
7.0
Ovarian ca.
4.7
0.0

IGROV-1

Stomach
0.0
0.0
Ovarian ca.
0.0
0.0

(ascites) SK-OV-3

Small intestine
0.0
0.0
Uterus
0.0
0.0

Colon ca. SW480
0.0
0.0
Placenta
0.0
13.0

Colon ca.* SW620
0.0
0.0
Prostate
2.6
0.0

(SW480 met)

Colon ca. HT29
0.0
15.6
Prostate ca.*
2.5
0.0

(bone met) PC-3

Colon ca. HCT-116
0.0
0.0
Testis
2.0
0.0

Colon ca. CaCo-2
0.0
0.0
Melanoma
0.0
0.0

Hs688(A).T

CC Well to Mod
0.0
14.3
Melanoma*
0.0
0.0

Diff (ODO3866)

(met)

Hs688(B).T

Colon ca. HCC-
0.0
0.0
Melanoma
0.0
0.0

2998

UACC-62

Gastric ca. (liver
0.0
100.0
Melanoma M14
0.0
0.0

met) NCI-N87

Bladder
0.8
49.3
Melanoma LOX
0.0
0.0

IMVI

Trachea
0.0
0.0
Melanoma*
0.0
0.0

(met) SK-MEL-5

Kidney
0.0
0.0
Adipose
0.0
0.0

[0518]

57

TABLE LD

Panel 2.2

Rel. Exp.

Rel. Exp.

(%) Ag2345,

(%) Ag2345,

Tissue Name
Run 174294755
Tissue Name
Run 174294755

Normal Colon
5.3
Kidney Margin (OD04348)
0.0

Colon cancer (OD06064)
0.0
Kidney malignant cancer
0.0

(OD06204B)

Colon Margin (OD06064)
0.0
Kidney normal adjacent
0.0

tissue (OD06204E)

Colon cancer (OD06159)
0.0
Kidney Cancer (OD04450-
0.0

01)

Colon Margin (OD06159)
0.0
Kidney Margin (OD04450-
0.0

03)

Colon cancer (OD06297-04)
0.0
Kidney Cancer 8120613
0.0

Colon Margin (OD06297-
0.0
Kidney Margin 8120614
0.0

015)

CC Gr.2 ascend colon
0.0
Kidney Cancer 9010320
0.0

(ODO3921)

CC Margin (ODO3921)
0.0
Kidney Margin 9010321
0.0

Colon cancer metastasis
0.0
Kidney Cancer 8120607
0.0

(OD06104)

Lung Margin (OD06104)
0.0
Kidney Margin 8120608
0.0

Colon mets to lung
0.0
Normal Uterus
0.0

(OD04451-01)

Lung Margin (OD04451-02)
0.0
Uterine Cancer 064011
6.8

Normal Prostate
18.2
Normal Thyroid
0.0

Prostate Cancer (OD04410)
6.5
Thyroid Cancer
0.0

Prostate Margin (OD04410)
100.0
Thyroid Cancer A302152
0.0

Normal Ovary
0.0
Thyroid Margin A302153
3.1

Ovarian cancer (OD06283-
0.0
Normal Breast
0.0

03)

Ovarian Margin (OD06283-
0.0
Breast Cancer
0.0

07)

Ovarian Cancer
24.3
Breast Cancer
15.6

Ovarian cancer (OD06145)
0.0
Breast Cancer (OD04590-
0.0

01)

Ovarian Margin (0D06145)
0.0
Breast Cancer Mets
0.0

(OD04590-03)

Ovarian cancer (OD06455-
0.0
Breast Cancer Metastasis
0.0

03)

Ovarian Margin (OD06455-
0.0
Breast Cancer
0.0

07)

Normal Lung
0.0
Breast Cancer 9100266
8.0

Invasive poor diff. lung
13.4
Breast Margin 9100265
0.0

adeno (ODO4945-01)

Lung Margin (ODO4945-03)
0.0
Breast Cancer A209073
0.0

Lung Malignant Cancer
7.4
Breast Margin A2090734
0.0

(OD03126)

Lung Margin (OD03126)
0.0
Breast cancer (OD06083)
0.0

Lung Cancer (OD05014A)
0.0
Breast cancer node
0.0

metastasis (OD06083)

Lung Margin (OD05014B)
5.2
Normal Liver
0.0

Lung cancer (OD06081)
0.0
Liver Cancer 1026
0.0

Lung Margin (OD06081)
0.0
Liver Cancer 1025
7.3

Lung Cancer (OD04237-01)
0.0
Liver Cancer 6004-T
0.0

Lung Margin (OD04237-02)
0.0
Liver Tissue 6004-N
0.0

Ocular Mel Met to Liver
0.0
Liver Cancer 6005-T
0.0

(ODO4310)

Liver Margin (ODO4310)
0.0
Liver Tissue 6005-N
0.0

Melanoma Metastasis
0.0
Liver Cancer
0.0

Lung Margin (OD04321)
0.0
Normal Bladder
0.0

Normal Kidney
0.0
Bladder Cancer
0.0

Kidney Ca, Nuclear grade 2
0.0
Bladder Cancer
0.0

(OD04338)

Kidney Margin (OD04338)
0.0
Normal Stomach
0.0

Kidney Ca Nuclear grade 1/2
0.0
Gastric Cancer 9060397
0.0

(OD04339)

Kidney Margin (OD04339)
0.0
Stomach Margin 9060396
0.0

Kidney Ca, Clear cell type
0.0
Gastric Cancer 9060395
0.0

(OD04340)

Kidney Margin (OD04340)
0.0
Stomach Margin 9060394
0.0

Kidney Ca, Nuclear grade 3
0.0
Gastric Cancer 064005
0.0

(OD04348)

[0519] Panel 1.3D Summary: Ag2494/Ag2345 Two experiments with two different probe and primer sets show highest expression of the CG50217-02 gene in a sample derived from lung cancer cell line and a gastric cancer cell line. This result suggests that expression of the CG50217-02 gene could be used to differentiate between lung and gastric cancer cell lines and other tissues and to detect the presence of gastric and lung cancers. Moreover, therapeutic modulation of the expression or function of this gene, through the use of small molecule drugs or antibodies, might be of use in the treatment of gastric and lung cancers.

[0520] Panel 2.2 Summary: Ag2345 Significant expression of the CG50217-02 gene is limited to normal prostate tissue adjacent to a prostate cancer (CT-33.5). The gene appears to be overexpressed in normal prostate tissue when compared to the adjacent tumor. Thus, therapeutic upregulation of the activity of the CG50217-02 gene product, through the application of the protein product or agonists might be of use in the treatment of prostate cancer

[0521] Panel 4D Summary: Ag2494/Ag2345 Expression of the CG50217-02 gene in this panel is low/undetectable (CT values>35) in all samples (data not shown).

[0522] M. CG55788-02/GMAC009758_B: Olfactory Receptor

[0523] Expression of gene CG55788-02 was assessed using the primer-probe sets Ag2319 and Ag2337, described in Tables MA and MB. Results of the RTQ-PCR runs are shown in Table MC.

58TABLE MAProbe Name Ag2319StartSEQ IDPrimersSequencesLengthPositionNO:Forward5′-attccacacaccttttgtgaac-3′22528219ProbeTET-5′-acattggcctagccaaatatgcatgt-3′-TAMRA26550220Reverse5′-ggaaaacccataccaaatgttt-3′22590221

[0524]

59

TABLE MB

Probe Name Ag2337

Start
SEQ ID

Primers
Sequences
Length
Position
NO:

Forward
5′-attccacacaccttttgtgaac-3′
22
528
222

Probe
TET-5′-acattggcctagccaaatatgcatgt-3′-TAMRA
26
550
223

Reverse
5′-ggaaaacccataccaaatgttt-3′
22
590
224

[0525]

60

TABLE MC

Panel 4.1D

Rel. Exp. (%)
Rel. Exp. (%)

Rel. Exp. (%)
Rel. Exp. (%)

Ag2319, Run
Ag2337, Run

Ag2319, Run
Ag2337, Run

Tissue Name
224781588
224781634
Tissue Name
224781588
224781634

Secondary Th1 act
0.0
0.0
HUVEC IL-1beta
0.0
0.0

Secondary Th2 act
0.0
0.0
HUVEC IFN gamma
0.0
0.0

Secondary Tr1 act
0.0
0.0
HUVEC TNF alpha +
0.0
0.0

IFN gamma

Secondary Th1 rest
0.0
0.0
HUVEC TNF alpha +
0.0
0.0

IL4

Secondary Th2 rest
0.0
0.0
HUVEC IL-11
0.0
0.0

Secondary Tr1 rest
0.0
0.0
Lung Microvascular
0.0
0.0

EC none

Primary Th1 act
0.0
0.0
Lung Microvascular
0.0
0.0

EC TNF alpha + IL-

1beta

Primary Th2 act
0.0
0.0
Microvascular
0.0
0.0

Dermal EC none

Primary Tr1 act
0.0
0.0
Microsvasular Dermal
0.0
0.0

EC TNF alpha + IL-

1beta

Primary Th1 rest
0.0
0.0
Bronchial epithelium
0.0
0.0

TNF alpha + IL1beta

Primary Th2 rest
0.0
0.0
Small airway
0.0
0.0

epithelium none

Primary Tr1 rest
0.0
0.0
Small airway
0.0
0.0

epithelium TNF alpha +

IL-1beta

CD45RA CD4
0.0
0.0
Coronery artery SMC
0.0
0.0

lymphocyte act

rest

CD45RO CD4
0.0
0.0
Coronery artery SMC
0.0
0.0

lymphocyte act

TNF alpha + IL-1beta

CD8 lymphocyte act
0.0
0.0
Astrocytes rest
0.0
0.0

Secondary CD8
0.0
0.0
Astrocytes TNF alpha +
0.0
0.0

lymphocyte rest

IL-1beta

Secondary CD8
0.0
0.0
KU-812 (Basophil)
0.0
0.0

lymphocyte act

rest

CD4 lymphocyte
0.0
0.0
KU-812 (Basophil)
0.0
0.0

none

PMA/ionomycin

2ry
0.0
0.0
CCD1106
0.0
0.0

Th1/Th2/Tr1_anti-

(Keratinocytes) none

CD95 CH11

LAK cells rest
0.0
0.0
CCD1106
0.0
0.0

(Keratinocytes)

TNF alpha + IL-1beta

LAK cells IL-2
0.0
0.0
Liver cirrhosis
0.0
0.0

LAK cells IL-2 + IL-
0.0
0.0
NCI-H292 none
0.0
0.0

12

LAK cells IL-2 + IFN
0.0
0.0
NCI-H292 IL-4
0.0
0.0

gamma

LAK cells IL-2 + IL-
0.0
0.0
NCI-H292 IL-9
0.0
0.0

18

LAK cells
0.0
0.0
NCI-H292 IL-13
0.0
0.0

PMA/ionomycin

NK Cells IL-2 rest
0.0
0.0
NCI-H292 IFN
0.0
0.0

gamma

Two Way MLR 3
0.0
0.0
HPAEC none
0.0
0.0

day

Two Way MLR 5
0.0
0.0
HPAEC TNF alpha +
0.0
0.0

day

IL-1beta

Two Way MLR 7
0.0
0.0
Lung fibroblast none
0.0
0.0

day

PBMC rest
0.0
0.0
Lung fibroblast TNF
0.0
0.0

alpha + IL-1beta

PBMC PWM
0.0
0.0
Lung fibroblast IL-4
0.0
0.0

PBMC PHA-L
0.0
0.0
Lung fibroblast IL-9
0.0
0.0

Ramos (B cell) none
0.0
0.0
Lung fibroblast IL-13
0.0
0.0

Ramos (B cell)
0.0
0.0
Lung fibroblast IFN
0.0
0.0

ionomycin

gamma

B lymphocytes PWM
0.0
0.0
Dermal fibroblast
0.0
0.0

CCD1070 rest

B lymphocytes
0.0
0.0
Dermal fibroblast
0.0
0.0

CD40L and IL-4

CCD1070 TNF alpha

EOL-1 dbcAMP
0.0
0.0
Dermal fibroblast
0.0
0.0

CCD1070 IL-1beta

EOL-1 dbcAMP
0.0
0.0
Dermal fibroblast IFN
0.0
1.0

PMA/ionomycin

gamma

Dendritic cells none
0.0
0.0
Dermal fibroblast IL-4
0.0
0.0

Dendritic cells LPS
0.0
0.0
Dermal Fibroblasts
0.0
0.0

rest

Dendritic cells anti-
0.0
0.0
Neutrophils
0.0
1.2

CD40

TNFa + LPS

Monocytes rest
0.0
0.0
Neutrophils rest
0.0
1.1

Monocytes LPS
0.0
0.0
Colon
0.0
0.0

Macrophages rest
0.0
0.0
Lung
1.7
1.8

Macrophages LPS
0.0
0.0
Thymus
9.6
14.5

HUVEC none
0.0
0.0
Kidney
100.0
100.0

HUVEC starved
0.0
0.0

[0526] CNS_neurodegeneration_v1.0 Summary: Ag2319/Ag2337 Expression of the CG55788-02 gene is low/undetectable (CTs>35) across all of the samples on this panel (data not shown).

[0527] General_screening_panel_v1.5 Summary: Ag2337 Data from one experiment indicates that there was a probable probe failure. (Data not shown.)

[0528] Panel 1.3D Summary: Ag2319/Ag2337 Data from two experiments indicates that there was a probable probe failure. (Data not shown.)

[0529] Panel 2.2 Summary: Ag2337 Data from one experiment indicates that there was a probable probe failure. (Data not shown.)

[0530] Panel 4.1D Summary: Ag2319/Ag2337 The CG55788-02 gene is only expressed at detectable levels in the kidney. The putative GPCR encoded for by this gene could allow cells within the kidney to respond to specific microenvironmental signals (For example, ref. 1). Therefore, antibody or small molecule therapies designed with the protein encoded for by this gene could modulate kidney function and be important in the treatment of inflammatory or autoimmune diseases that affect the kidney, including lupus and glomerulonephritis (Mark et al., G protein modulation of recombinant P/Q-type calcium channels by regulators of G protein signalling proteins. J. Physiol. 528 Pt 1: 65-77, 2000).

[0531] Panel 4D Summary: Ag2319/Ag2337 Expression of this gene is low/undetectable (CTs>35) in all of the tissues on this panel (data not shown).

[0532] N. CG149194-01/GMAC023106_C: Olfactory Receptor

[0533] Expression of gene CG149194-01 was assessed using the primer-probe set Ag2325, described in Table NA. Results of the RTQ-PCR runs are shown in Tables NB and NC.

61TABLE NAProbe Name Ag2325StartSEQ IDPrimersSequencesLengthPositionNO:Forward5′-cagtcattgctgagttcatcct-3′2266225ProbeTET-5′-ctgcagccagttgtctttgtgctcct-3′-TAMRA26113226Reverse5′-cgtgaccaggtaggcaaag-3′19142227

[0534]

62

TABLE NB

CNS_neurodegeneration_v1.0

Rel. Exp.

Rel. Exp.

(%) Ag2325, Run

(%) Ag2325, Run

Tissue Name
207929054
Tissue Name
207929054

AD 1 Hippo
16.4
Control (Path) 3
4.2

Temporal Ctx

AD 2 Hippo
16.5
Control (Path) 4
27.7

Temporal Ctx

AD 3 Hippo
8.2
AD 1 Occipital Ctx
7.5

AD 4 Hippo
10.2
AD 2 Occipital Ctx
0.0

(Missing)

AD 5 hippo
59.5
AD 3 Occipital Ctx
7.5

AD 6 Hippo
23.5
AD 4 Occipital Ctx
7.5

Control 2 Hippo
10.5
AD 5 Occipital Ctx
14.4

Control 4 Hippo
8.6
AD 6 Occipital Ctx
4.4

Control (Path) 3 Hippo
13.0
Control 1 Occipital Ctx
8.5

AD 1 Temporal Ctx
23.3
Control 2 Occipital Ctx
16.8

AD 2 Temporal Ctx
9.9
Control 3 Occipital Ctx
3.4

AD 3 Temporal Ctx
2.6
Control 4 Occipital Ctx
4.5

AD 4 Temporal Ctx
18.4
Control (Path) 1
34.9

Occipital Ctx

AD 5 Inf Temporal Ctx
100.0
Control (Path) 2
7.3

Occipital Ctx

AD 5 SupTemporal Ctx
23.2
Control (Path) 3
7.0

Occipital Ctx

AD 6 Inf Temporal Ctx
28.7
Control (Path) 4
17.1

Occipital Ctx

AD 6 Sup Temporal Ctx
18.2
Control 1 Parietal Ctx
26.2

Control 1 Temporal Ctx
15.1
Control 2 Parietal Ctx
23.5

Control 2 Temporal Ctx
4.5
Control 3 Parietal Ctx
7.7

Control 3 Temporal Ctx
6.6
Control (Path) 1
32.3

Parietal Ctx

Control 4 Temporal Ctx
2.9
Control (Path) 2
8.2

Parietal Ctx

Control (Path) 1
13.1
Control (Path) 3
5.8

Temporal Ctx

Parietal Ctx

Control (Path) 2
14.7
Control (Path) 4
2.9

Temporal Ctx

Parietal Ctx

[0535]

63

TABLE NC

Panel 4D

Rel. Exp.

Rel. Exp.

(%) Ag2325,

(%) Ag2325,

Tissue Name
Run 164022571
Tissue Name
Run 164022571

Secondary Th1 act
5.0
HUVEC IL-1beta
0.0

Secondary Th2 act
14.9
HUVEC IFN gamma
0.0

Secondary Tr1 act
0.0
HUVEC TNF alpha + IFN
1.8

gamma

Secondary Th1 rest
0.0
HUVEC TNF alpha + IL4
11.5

Secondary Th2 rest
20.7
HUVEC IL-11
0.0

Secondary Tr1 rest
15.4
Lung Microvascular EC none
0.0

Primary Th1 act
29.1
Lung Microvascular EC
7.9

TNF alpha + IL-1beta

Primary Th2 act
35.6
Microvascular Dermal EC none
0.0

Primary Tr1 act
56.6
Microsvasular Dermal EC
0.0

TNF alpha + IL-1beta

Primary Th1 rest
14.8
Bronchial epithelium TNF
0.0

alpha + IL1beta

Primary Th2 rest
22.4
Small airway epithelium none
0.0

Primary Tr1 rest
25.7
Small airway epithelium
5.1

TNF alpha + IL-1beta

CD45RA CD4 lymphocyte
11.1
Coronery artery SMC rest
0.0

act

CD45RO CD4 lymphocyte
31.9
Coronery artery SMC TNF
0.0

act

alpha + IL-1beta

CD8 lymphocyte act
5.6
Astrocytes rest
0.0

Secondary CD8
21.2
Astrocytes TNF alpha + IL-1beta
0.0

lymphocyte rest

Secondary CD8
0.0
KU-812 (Basophil) rest
0.0

lymphocyte act

CD4 lymphocyte none
5.6
KU-812 (Basophil)
0.0

PMA/ionomycin

2ry Th1/Th2/Tr1_anti-
16.0
CCD1106 (Keratinocytes) none
22.7

CD95 CH11

LAK cells rest
4.3
CCD1106 (Keratinocytes)
4.9

TNF alpha + IL-1beta

LAK cells IL-2
0.0
Liver cirrhosis
15.4

LAK cells IL-2 + IL-12
24.5
Lupus kidney
0.0

LAK cells IL-2 + IFN
13.8
NCI-H292 none
0.0

gamma

LAK cells IL-2 + IL-18
30.4
NCI-H292 IL-4
3.8

LAK cells
0.0
NCI-H292 IL-9
0.0

PMA/ionomycin

NK cells IL-2 rest
5.6
NCI-H292 IL-13
0.0

Two Way MLR 3 day
0.0
NCI-H292 IFN gamma
0.0

Two Way MLR 5 day
6.5
HPAEC none
0.0

Two Way MLR 7 day
0.0
HPAEC TNF alpha + IL-1beta
0.0

PBMC rest
0.0
Lung fibroblast none
4.9

PBMC PWM
50.7
Lung fibroblast TNF alpha + IL-
5.7

1beta

PBMC PHA-L
26.8
Lung fibroblast IL-4
5.4

Ramos (B cell) none
0.0
Lung fibroblast IL-9
5.6

Ramos (B cell) ionomycin
0.0
Lung fibroblast IL-13
4.0

B lymphocytes PWM
31.0
Lung fibroblast IFN gamma
6.5

B lymphocytes CD40L
12.2
Dermal fibroblast CCD1070 rest
14.2

and IL-4

EOL-1 dbcAMP
100.0
Dermal fibroblast CCD1070
5.3

TNF alpha

EOL-1 dbcAMP
63.3
Dermal fibroblast CCD1070 IL-
0.0

PMA/ionomycin

1beta

Dendritic cells none
0.0
Dermal fibroblast IFN gamma
0.0

Dendritic cells LPS
0.0
Dermal fibroblast IL-4
3.3

Dendritic cells anti-CD40
11.6
IBD Colitis 2
0.0

Monocytes rest
0.0
IBD Crohn's
8.3

Monocytes LPS
0.0
Colon
2.3

Macrophages rest
0.0
Lung
0.0

Macrophages LPS
0.0
Thymus
4.8

HUVEC none
0.0
Kidney
44.8

HUVEC starved
10.4

[0536] CNS_neurodegeneration_v1.0 Summary: Ag2325 The CG149194-01 gene represents a novel G-protein coupled receptor (GPCR) with expression in the brain, as seen in this panel. The GPCR family of receptors contains a large number of neurotransmitter receptors, including the dopamine, serotonin, a and b-adrenergic, acetylcholine muscarinic, histamine, peptide, and metabotropic glutamate receptors. GPCRs are excellent drug targets in various neurologic and psychiatric diseases. All antipsychotics have been shown to act at the dopamine D2 receptor; similarly novel antipsychotics also act at the serotonergic receptor, and often the muscarinic and adrenergic receptors as well. While the majority of antidepressants can be classified as selective serotonin reuptake inhibitors, blockade of the 5-HT1A and a2 adrenergic receptors increases the effects of these drugs. The GPCRs are also of use as drug targets in the treatment of stroke. Blockade of the glutamate receptors may decrease the neuronal death resulting from excitotoxicity; further more the purinergic receptors have also been implicated as drug targets in the treatment of cerebral ischemia. The b-adrenergic receptors have been implicated in the treatment of ADHD with Ritalin, while the a-adrenergic receptors have been implicated in memory. Therefore this gene may be of use as a small molecule target for the treatment of any of the described diseases (El Yacoubi et al., Adenosine A2A receptor antagonists are potential antidepressants: evidence based on pharmacology and A2A receptor knockout mice. Br J Pharmacol 134(1):68-77, 2001; Blier, Pharmacology of rapid-onset antidepressant treatment strategies. Clin Psychiatry 62 Suppl 15:12-7, 2001; Tranquillini and Reggiani, Glycine-site antagonists and stroke. Expert Opin Investig Drugs 8(11):1837-1848, 1999; Monopoli et al., Blockade of adenosine A2A receptors by SCH 58261 results in neuroprotective effects in cerebral ischaemia in rats. Neuroreport 9(17):3955-9, 1998).

[0537] Panel 1.3D Summary: Ag2325 Expression of the CG149194-01 gene is low/undetectable (CTs>35) across all of the samples on this panel (data not shown).

[0538] Panel 2.2 Summary: Ag2325 Expression of the CG149194-01 gene is low/undetectable (CTs>35) across all of the samples on this panel (data not shown).

[0539] Panel 4D Summary: Ag2325 The highest level of expression of the CG149194-01 is detected in resting EOL-1 eosinophil cells (CT=32.3). Lower but still significant levels of expression are found in T cells including activated primary Th1, Th2 and Tr1 cells, resting primary Th2 and Tr1 cells, CD45RO CD4 lymphocytes, resting secondary CD8 lymphocytes, and IL2+IL12 and IL2+IL18 stimulated lymphokine activated killer (LAK) cells. Additional low level expression is detected in peripheral blood mononuclear cells (PBMC), pokeweed mitogen stimulated B lymphocytes, resting CCD1106 keratinocytes, and normal kidney. Since eosinophils and T cells play an important role in lung pathology, inflammatory bowel disease and autoimmune disorders including rheumatoid arthritis, antibody or small molecule therapies designed with the protein encoded by this gene could block or inhibit inflammation or tissue damage due to lung conditions including asthma, allergies, hypersensitivity reactions, inflammatory bowel disease, viral infections and autoimmune disease. The expression of this gene in T cells, B cells, and PBMCs also suggests that therapeutic modulation of the gene product may ameliorate symptoms of synovitis associated with osteoarthritis. Detection of this gene in LAK cells suggests that modulation of the function of the protein encoded by this gene with a small molecule drug or antibody may lead to improvement of symptoms associated with tumor immunology and tumor cell clearance, as well as removal of virally and bacterial infected cells.

[0540] O. CG56715-02/GMAC026083_E: Olfactory Receptor

[0541] Expression of gene CG56715-02 was assessed using the primer-probe set Ag2619, described in Table OA. Results of the RTQ-PCR runs are shown in Tables OB, OC, and OD.

64TABLE OAProbe Name Ag2619SEQ IDPrimersSequencesLengthStartPositionNO:Forward5′-acgttcgtagaatcagctctga-3′22271228ProbeTET-5′-tcttccttcatggattctcctttatgg-3′-TAMRA27313229Reverse5′-atagccaggaggacagaagact-3′22340230

[0542]

65

TABLE OB

Panel 1.3D

Rel. Exp.

Rel. Exp.

(%) Ag2619, Run

(%) Ag2619, Run

Tissue Name
167644079
Tissue Name
167644079

Liver adenocarcinoma
2.1
Kidney (fetal)
2.0

Pancreas
0.0
Renal ca.786-0
0.0

Pancreatic ca. CAPAN 2
0.0
Renal ca. A498
0.0

Adrenal gland
0.0
Renal ca. RXF 393
0.7

Thyroid
0.6
Renal ca. ACHN
0.4

Salivary gland
0.0
Renal ca. UO-31
0.0

Pituitary gland
0.0
Renal ca. TK-10
0.0

Brain (fetal)
1.8
Liver
0.0

Brain (whole)
1.9
Liver (fetal)
0.0

Brain (amygdala)
0.0
Liver ca. (hepatoblast)
0.6

HepG2

Brain (cerebellum)
1.5
Lung
0.0

Brain (hippocampus)
0.6
Lung (fetal)
0.0

Brain (substantia nigra)
0.0
Lung ca. (small cell) LX-1
39.0

Brain (thalamus)
0.0
Lung ca. (small cell)
0.0

NCI-H69

Cerebral Cortex
0.0
Lung ca. (s. cell var.)
7.6

SHP-77

Spinal cord
2.0
Lung ca. (large cell)NCI-
0.0

H460

glio/astro U87-MG
0.0
Lung ca. (non-sm. cell)
1.7

A549

glio/astro U-118-MG
0.6
Lung ca. (non-s. cell)
0.0

NCI-H23

astrocytoma SW1783
0.0
Lung ca. (non-s. cell)
0.0

HOP-62

neuro*; met SK-N-AS
0.0
Lung ca. (non-c. cl) NCI-
0.0

H522

astrocytoma SF-539
0.0
Lung ca. (squam.) SW
0.7

900

astrocytoma SNB-75
0.0
Lung ca. (squam.) NCI-
0.0

H596

glioma SNB-19
0.0
Mammary gland
0.6

glioma U251
0.6
Breast ca.* (pl. ef) MCF-7
0.0

glioma SF-295
0.0
Breast ca.* (pl. ef) MDA-
0.0

MB-231

Heart (fetal)
0.0
Breast ca.* (pl. ef) T47D
0.0

Heart
0.0
Breast ca. BT-549
0.8

Skeletal muscle (fetal)
0.0
Breast ca. MDA-N
13.2

Skeletal muscle
0.0
Ovary
0.0

Bone marrow
0.0
Ovarian ca. OVCAR-3
1.2

Thymus
1.4
Ovarian ca. OVCAR-4
0.0

Spleen
0.0
Ovarian ca. OVCAR-5
1.0

Lymph node
0.0
Ovarian ca. OVCAR-8
0.0

Colorectal
0.4
Ovarian ca. IGROV-1
0.0

Stomach
0.0
Ovarian ca.* (ascites)
5.4

SK-OV-3

Small intestine
0.0
Uterus
1.6

Colon ca. SW480
0.0
Plancenta
0.4

Colan ca.* SW620(SW480
8.0
Prostate
0.0

met)

Colon ca. HT29
0.0
Prostate ca.* (bone
0.0

met)PC-3

Colon ca. HCT-116
100.0
Testis
0.0

Colon ca. CaCo-2
0.0
Melanoma Hs688(A).T
0.0

Colon ca.
0.0
Melanoma* (met)
0.0

tissue(ODO3866)

Hs688(B).T

Colon ca. HCC-2998
0.0
Melanoma UACC-62
0.0

Gastric ca.* (liver met)
0.0
Melanoma M14
1.4

NCI-N87

Bladder
0.9
Melanoma LOX IMVI
7.6

Trachea
0.0
Melanoma* (met) SK-
48.6

MEL-5

Kidney
0.0
Adipose
0.7

[0543]

66

TABLE OC

Panel 2.2

Rel. Exp.

Rel. Exp.

(%) Ag2619,

(%) Ag2619,

Tissue Name
Run 175135477
Tissue Name
Run 175135477

Normal Colon
0.0
Kidney Margin (OD04348)
71.2

Colon cancer (OD06064)
0.0
Kidney malignant cancer
0.0

(OD06204B)

Colon Margin (OD06064)
0.0
Kidney normal adjacent
0.0

tissue (OD06204E)

Colon cancer (OD06159)
0.0
Kidney Cancer (OD04450-
5.0

01)

Colon Margin (OD06159)
0.0
Kidney Margin (OD04450-
7.6

03)

Colon cancer (OD06297-04)
0.0
Kidney Cancer 8120613
0.0

Colon Margin (OD06297-
0.0
Kidney Margin 8120614
0.0

015)

CC Gr.2 ascend colon
0.0
Kidney Cancer 9010320
0.0

(ODO3921)

CC Margin (ODO3921)
0.0
Kidney Margin 9010321
4.7

Colon cancer metastasis
0.0
Kidney Cancer 8120607
0.0

(OD06104)

Lung Margin (OD06104)
0.0
Kidney Margin 8120608
0.0

Colon mets to lung
3.5
Normal Uterus
5.9

(OD04451-01)

Lung Margin (OD04451-02)
2.3
Uterine Cancer 064011
4.0

Normal Prostate
3.5
Normal Thyroid
0.0

Prostate Cancer (OD04410)
0.0
Thyroid Cancer 064010
0.0

Prostate Margin (OD04410)
0.0
Thyroid Cancer A302152
14.9

Normal Ovary
0.0
Thyroid Margin A302153
0.0

Ovarian cancer (OD06283-
0.0
Normal Breast
0.0

03)

Ovarian Margin (OD06283-
3.0
Breast Cancer (OD04566)
0.0

07)

Ovarian Cancer 064008
15.7
Breast Cancer 1024
1.7

Ovarian cancer (OD06145)
0.0
Breast Cancer (OD04590-
4.0

01)

Ovarian Margin (OD06145)
3.5
Breast Cancer Mets
0.0

(OD04590-03)

Ovarian cancer (OD06455-
0.0
Breast Cancer Metastasis
0.0

03)

(OD04655-05)

Ovarian Margin (OD06455-
1.7
Breast Cancer 064006
0.0

07)

Normal Lung
0.0
Breast Cancer 9100266
10.3

Invasive poor diff. lung
100.0
Breast Margin 9100265
0.0

adeno (ODO4945-01)

Lung Margin (ODO4945-03)
0.0
Breast Cancer A209073
0.0

Lung Malignant Cancer
2.8
Breast Margin A2090734
9.4

(OD03126)

Lung Margin (OD03126)
0.0
Breast cancer (OD06083)
3.2

Lung Cancer (OD05014A)
0.0
Breast cancer node
0.0

metastasis (OD06083)

Lung Margin (OD05014B)
3.1
Normal Liver
0.0

Lung cancer (OD06081)
0.0
Liver Cancer 1026
0.0

Lung Margin (OD06081)
4.7
Liver Cancer 1025
14.1

Lung Cancer (OD04237-01)
7.3
Liver Cancer 6004-T
0.0

Lung Margin (OD04237-02)
0.0
Liver Tissue 6004-N
0.0

Ocular Melanoma Metastasis
0.0
Liver Cancer 6005-T
0.0

Ocular Melanoma Margin
4.7
Liver Tissue 6005-N
0.0

(Liver)

Melanoma Metastasis
0.0
Liver Cancer 064003
3.8

Melanoma Margin (Lung)
7.9
Normal Bladder
9.8

Normal Kidney
4.6
Bladder Cancer 1023
0.0

Kidney Ca, Nuclear grade 2
30.1
Bladder Cancer A302173
0.0

(OD04338)

Kidney Margin (OD04338)
0.0
Normal Stomach
5.3

Kidney Ca Nuclear grade 1/2
38.7
Gastric Cancer 9060397
0.0

(OD04339)

Kidney Margin (OD04339)
0.0
Stomach Margin 9060396
0.0

Kidney Ca, Clear cell type
0.0
Gastric Cancer 9060395
0.0

(OD04340)

Kidney Margin (OD04340)
5.1
Stomach Margin 9060394
0.0

Kidney Ca, Nuclear grade 3
0.0
Gastric Cancer 064005
0.0

(OD04348)

[0544]

67

TABLE OD

Panel 4D

Rel. Exp.

Rel. Exp.

(%) Ag2619,

(%) Ag2619,

Tissue Name
Run 164401285
Tissue Name
Run 164401285

Secondary Th1 act
9.7
HUVEC IL-1beta
0.0

Secondary Th2 act
0.0
HUVEC IFN gamma
0.0

Secondary Tr1 act
0.0
HUVEC TNF alpha + IFN
0.0

gamma

Secondary Th1 rest
0.0
HUVEC TNF alpha + IL4
0.0

Secondary Th2 rest
0.0
HUVEC IL-11
0.0

Secondary Tr1 rest
0.0
Lung Microvascular EC none
4.4

Primary Th1 act
0.0
Lung Microvascular EC
0.0

TNF alpha + IL-1beta

Primary Th2 act
0.0
Microvascular Dermal EC none
1.6

Primary Tr1 act
0.0
Microsvasular Dermal EC
0.0

TNF alpha + IL-1beta

Primary Th1 rest
0.0
Bronchial epithelium TNF
0.0

alpha + IL1beta

Primary Th2 rest
0.0
Small airway epithelium none
0.0

Primary Tr1 rest
0.8
Small airway epithelium
2.6

TNF alpha + IL-1beta

CD45RA CD4 lymphocyte
0.0
Coronery artery SMC rest
0.0

act

CD45RO CD4 lymphocyte
0.0
Coronery artery SMC TNF
0.0

act

alpha + IL-1beta

CD8 lymphocyte act
2.5
Astrocytes rest
0.0

Secondary CD8
1.6
Astrocytes TNF alpha + IL-1beta
0.0

lymphocyte rest

Secondary CD8
1.7
KU-812 (Basophil) rest
0.0

lymphocyte act

CD4 lymphocyte none
0.0
KU-812 (Basophil)
0.0

PMA/ionomycin

2ry Th1/Th2/Tr1_anti-
0.0
CCD1106 (Keratinocytes) none
0.0

CD95 CH11

LAK cells rest
2.0
CCD1106 (Keratinocytes)
0.0

TNF alpha + IL-1beta

LAK cells IL-2
0.0
Liver cirrhosis
9.4

LAK cells IL-2 + IL-12
0.0
Lupus kidney
6.0

LAK cells IL-2 + IFN
0.0
NCI-H292 none
0.0

gamma

LAK cells IL-2 + IL-18
0.0
NCI-H292 IL-4
0.0

LAK cells
1.8
NCI-H292 IL-9
0.0

PMA/ionomycin

NK Cells IL-2 rest
0.0
NCI-H292 IL-13
0.0

Two Way MLR 3 day
5.0
NCI-H292 IFN gamma
0.0

Two Way MLR 5 day
0.0
HPAEC none
0.0

Two Way MLR 7 day
0.0
HPAEC TNF alpha + IL-1beta
0.0

PBMC rest
0.0
Lung fibroblast none
0.0

PBMC PWM
0.0
Lung fibroblast TNF alpha + IL-
0.0

1beta

PBMC PHA-L
0.0
Lung fibroblast IL-4
2.5

Ramos (B cell) none
37.9
Lung fibroblast IL-9
1.6

Ramos (B cell) ionomycin
100.0
Lung fibroblast IL-13
0.0

B lymphocytes PWM
1.4
Lung fibroblast IFN gamma
0.0

B lymphocytes CD40L
2.8
Dermal fibroblast CCD1070 rest
0.0

and IL-4

EOL-1 dbcAMP
0.0
Dermal fibroblast CCD1070
2.0

TNF alpha

EOL-1 dbcAMP
0.0
Dermal fibroblast CCD1070 IL-
0.0

PMA/ionomycin

1beta

Dendritic cells none
0.0
Dermal fibroblast IFN gamma
0.0

Dendritic cells LPS
2.3
Dermal fibroblast IL-4
0.0

Dendritic cells anti-CD40
0.0
IBD Colitis 2
3.6

Monocytes rest
0.0
IBD Crohn's
1.9

Monocytes LPS
0.0
Colon
0.0

Macrophages rest
0.0
Lung
0.0

Macrophages LPS
0.0
Thymus
17.7

HUVEC none
0.0
Kidney
3.4

HUVEC starved
0.0

[0545] Panel 1.3D Summary: Ag2619 The CG56715-02 gene shows highest expression in a sample derived from a colon cancer cell line (HCT116)(CT=31.4). In addition, there is substantial expression in a melanoma cell line (SK-MEL-5) and a lung cancer cell line (LX-1). Thus, this gene could be used to distinguish these cell lines form other samples in the panel. Moreover, therapeutic modulation of this gene, through the use of small molecule drugs, antibodies or protein therapeutics might be of use in the treatment of melanoma, lung cancer or colon cancer.

[0546] Panel 2.2 Summary: Ag2619 The expression of the CG56715-02 gene is highest in a sample derived from a poorly differentiated lung cancer (CT=33.8). In addition, there is substantial expression in a sample of normal kidney adjacent to a kidney malignancy. Thus, the expression of this gene could be used to distinguish these tow samples from the other samples in the panel. Moreover, therapeutic modulation of this gene, through the use of small molecule drugs, antibodies or protein therapeutics might be of benefit in the treatment of lung cancer.

[0547] Panel 4D Summary: Ag2619 The highest expression of the CG56715-02 (CT=32.8) is detected in Ramos B cells stimulated with ionomycin. Lower but still significant levels of expression are seen in untreated Ramos B cells (CT=34.16). B cells represent a principle component of immunity and contribute to the immune response in a number of important functional roles, including antibody production. Production of antibodies against self-antigens is a major component of autoimmune disorders. Since these cells are important in inflammatory processes and inflammatory cascades, therapeutic modulation of this gene product may reduce or eliminate the symptoms of patients suffering from asthma, allergies, chronic obstructive pulmonary disease, emphysema, Crohn's disease, ulcerative colitis, rheumatoid arthritis, psoriasis, osteoarthritis, and other autoimmune disorders.

[0548] P. CG56720-01/GMAC026083_D: Olfactory Receptor

[0549] Expression of gene CG56720-01 was assessed using the primer-probe set Ag2618, described in Table PA. Results of the RTQ-PCR runs are shown in Tables PB, PC and PD.

68TABLE PAProbe Name Ag2618SEQ IDPrimersSequencesLengthStartPositionNO:Forward5′-cgctttgtggctatttgttatc-3′22389231ProbeTET-5′-cgctatgtcactgtgctcactcacaa-3′-TAMRA26416232Reverse5′-cccagacccatagccaatatac-3′22444234

[0550]

69

TABLE PB

Panel 1.3D

Rel. Exp.

Rel. Exp.

(%) Ag2618, Run

(%) Ag2618, Run

Tissue Name
167644790
Tissue Name
167644790

Liver adenocarcinoma
0.0
Kidney (fetal)
2.1

Pancreas
0.3
Renal ca.786-0
0.3

Pancreatic ca. CAPAN 2
0.0
Renal ca. A498
0.0

Adrenal gland
0.4
Renal ca. RXF 393
0.5

Thyroid
0.0
Renal ca. ACHN
0.0

Salivary gland
0.3
Renal ca. UO-31
0.0

Pituitary gland
0.0
Renal ca. TK-10
0.3

Brain (fetal)
0.3
Liver
0.0

Brain (whole)
0.0
Liver (fetal)
0.0

Brain (amygdala)
0.3
Liver ca. (hepatoblast)
0.3

HepG2

Brain (cerebellum)
0.0
Lung
0.3

Brain (hippocampus)
0.0
Lung (fetal)
1.0

Brain (substantia nigra)
0.0
Lung ca. (small cell) LX-1
28.1

Brain (thalamus)
0.0
Lung ca. (small cell)
0.0

NCI-H69

Cerebral Cortex
0.0
Lung ca. (s. cell var.)
2.7

SHP-77

Spinal cord
0.0
Lung ca. (large cell)NCI-
0.0

H460

glio/astro U87-MG
0.0
Lung ca. (non-sm. cell)
0.4

A549

glio/astro U-118-MG
0.6
Lung ca. (non-s. cell)
0.0

NCI-H23

astrocytoma SW1783
0.2
Lung ca. (non-s. cell)
0.0

HOP-62

neuro*; met SK-N-AS
0.0
Lung ca. (non-c. cl) NCI-
0.0

H522

astrocytoma SF-539
1.0
Lung ca. (squam.) SW
0.0

900

astrocytoma SNB-75
0.0
Lung ca. (squam.) NCI-
0.0

H596

glioma SNB-19
0.0
Mammary gland
0.0

glioma U251
0.6
Breast ca.* (pl. ef) MCF-7
0.0

glioma SF-295
1.2
Breast ca.* (pl. ef) MDA-
0.0

MB-231

Heart (fetal)
0.3
Breast ca.* (pl. ef) T47D
0.0

Heart
0.0
Breast ca. BT-549
0.0

Skeletal muscle (fetal)
0.5
Breast ca. MDA-N
12.0

Skeletal muscle
0.0
Ovary
0.0

Bone marrow
0.0
Ovarian ca. OVCAR-3
0.7

Thymus
0.0
Ovarian ca. OVCAR-4
0.0

Spleen
0.0
Ovarian ca. OVCAR-5
0.5

Lymph node
0.5
Ovarian ca. OVCAR-8
0.0

Colorectal
0.5
Ovarian ca. IGROV-1
0.0

Stomach
0.0
Ovarian ca.* (ascites)
4.9

SK-OV-3

Small intestine
0.0
Uterus
0.0

Colon ca. SW480
0.2
Plancenta
0.0

Colan ca.* SW620(SW480
8.4
Prostate
0.0

met)

Colon ca. HT29
0.0
Prostate ca.* (bone
0.0

met)PC-3

Colon ca. HCT-116
100.0
Testis
0.0

Colon ca. CaCo-2
0.0
Melanoma Hs688(A).T
0.0

Colon ca.
0.0
Melanoma* (met)
0.0

tissue(ODO3866)

Hs688(B).T

Colon ca. HCC-2998
0.0
Melanoma UACC-62
0.0

Gastric ca.* (liver met)
0.7
Melanoma M14
0.3

NCI-N87

Bladder
0.9
Melanoma LOX IMVI
10.9

Trachea
0.0
Melanoma* (met) SK-
38.2

MEL-5

Kidney
0.5
Adipose
0.3

[0551]

70

TABLE PC

Panel 2.2

Rel. Exp. (%)

Rel. Exp. (%)

Ag2618,

Ag2618,

Run

Run

Tissue Name
175063680
Tissue Name
175063680

Normal Colon
0.0
Kidney Margin (OD04348)
100.0

Colon cancer (OD06064)
6.3
Kidney malignant cancer
0.0

(OD06204B)

Colon Margin (OD06064)
0.0
Kidney normal adjacent
0.0

tissue (OD06204E)

Colon cancer (OD06159)
0.0
Kidney Cancer (OD04450-
24.7

01)

Colon Margin (OD06159)
10.1
Kidney Margin (OD04450-
11.8

03)

Colon cancer (OD06297-04)
0.0
Kidney Cancer 8120613
0.0

Colon Margin (OD06297-
9.1
Kidney Margin 8120614
0.0

015)

CC Gr.2 ascend colon
17.8
Kidney Cancer 9010320
0.0

(ODO3921)

CC Margin (ODO3921)
0.0
Kidney Margin 9010321
0.0

Colon cancer metastasis
0.0
Kidney Cancer 8120607
0.0

(OD06104)

Lung Margin (OD06104)
0.0
Kidney Margin 8120608
0.0

Colon mets to lung
0.0
Normal Uterus
16.8

(OD04451-01)

Lung Margin (OD04451-02)
0.0
Uterine Cancer 064011
0.0

Normal Prostate
0.0
Normal Thyroid
0.0

Prostate Cancer (OD04410)
0.0
Thyroid Cancer 064010
0.0

Prostate Margin (OD04410)
0.0
Thyroid Cancer A302152
12.2

Normal Ovary
0.0
Thyroid Margin A302153
0.0

Ovarian cancer (OD06283-
0.0
Normal Breast
13.2

03)

Ovarian Margin (OD06283-
10.0
Breast Cancer (OD04566)
0.0

07)

Ovarian Cancer 064008
76.3
Breast Cancer 1024
0.0

Ovarian cancer (OD06145)
0.0
Breast Cancer (OD04590-
0.0

01)

Ovarian Margin (OD06145)
19.5
Breast Cancer Mets
19.1

(OD04590-03)

Ovarian cancer (OD06455-
0.0
Breast Cancer Metastasis
21.3

03)

(OD04655-05)

Ovarian Margin (OD06455-
0.0
Breast Cancer 064006
3.6

07)

Normal Lung
14.9
Breast Cancer 9100266
0.0

Invasive poor diff. lung
15.0
Breast Margin 9100265
0.0

adeno (ODO4945-01

Lung Margin (ODO4945-03)
8.6
Breast Cancer A209073
0.0

Lung Malignant Cancer
0.0
Breast Margin A2090734
0.0

(OD03126)

Lung Margin (OD03126)
8.2
Breast cancer (OD06083)
0.0

Lung Cancer (OD05014A)
0.0
Breast cancer node
0.0

metastasis (OD06083)

Lung Margin (OD05014B)
6.8
Normal Liver
12.6

Lung cancer (OD06081)
0.0
Liver Cancer 1026
0.0

Lung Margin (OD06081)
0.0
Liver Cancer 1025
5.4

Lung Cancer (OD04237-01)
0.0
Liver Cancer 6004-T
0.0

Lung Margin (OD04237-02)
0.0
Liver Tissue 6004-N
0.0

Ocular Melanoma Metastasis
0.0
Liver Cancer 6005-T
0.0

Ocular Melanoma Margin
0.0
Liver Tissue 6005-N
0.0

(Liver)

Melanoma Metastasis
0.0
Liver Cancer 064003
0.0

Melanoma Margin (Lung)
0.0
Normal Bladder
0.0

Normal Kidney
7.2
Bladder Cancer 1023
0.0

Kidney Ca, Nuclear grade 2
31.0
Bladder Cancer A302173
0.0

(OD04338)

Kidney Margin (OD04338)
14.3
Normal Stomach
12.2

Kidney Ca Nuclear grade 1/2
11.0
Gastric Cancer 9060397
0.0

(OD04339)

Kidney Margin (OD04339)
14.2
Stomach Margin 9060396
0.0

Kidney Ca, Clear cell type
10.2
Gastric Cancer 9060395
0.0

(OD04340)

Kidney Margin (OD04340)
5.3
Stomach Margin 9060394
0.0

Kidney Ca, Nuclear grade 3
0.0
Gastric Cancer 064005
0.0

(OD04348)

[0552]

71

TABLE PD

Panel 4D

Rel. Exp. (%)

Rel. Exp. (%)

Ag2618,

Ag2618,

Run

Run

Tissue Name
164299474
Tissue Name
164299474

Secondary Th1 act
0.0
HUVEC IL-1beta
0.0

Secondary Th2 act
0.0
HUVEC IFN gamma
0.0

Secondary Tr1 act
0.0
HUVEC TNFalpha + IFN
0.0

gamma

Secondary Th1 rest
0.0
HUVEC TNFalpha + IL4
0.0

Secondary Th2 rest
0.0
HUVEC IL-11
0.0

Secondary Tr1 rest
1.8
Lung Microvascular EC none
0.0

Primary Th1 act
0.0
Lung Microvascular EC
0.0

TNFalpha + IL-1beta

Primary Th2 act
0.0
Microvascular Dermal EC none
0.0

Primary Tr1 act
0.0
Microsvasular Dermal EC
0.0

TNFalpha + IL-1beta

Primary Th1 rest
4.8
Bronchial epithelium
0.0

TNF alpha + IL1beta

Primary Th2 rest
1.2
Small airway epithelium none
0.0

Primary Tr1 rest
1.1
Small airway epithelium
0.5

TNFalpha + IL-1beta

CD45RA CD4 lymphocyte
1.0
Coronery artery SMC rest
0.0

act

CD45RO CD4 lymphocyte
0.0
Coronery artery SMC
0.0

act

TNF alpha + IL-1beta

CD8 lymphocyte act
0.0
Astrocytes rest
0.0

Secondary CD8
1.2
Astrocytes TNFalpha + IL-1beta
0.0

lymphocyte rest

Secondary CD8
0.0
KU-812 (Basophil) rest
0.0

lymphocyte act

CD4 lymphocyte none
0.0
KU-812 (Basophil)
1.2

PMA/ionomycin

2ry Th1/Th2/Tr1_anti-
0.0
CCD1106 (Keratinocytes) none
0.0

CD95 CH11

LAK cells rest
0.0
CCD1106 (Keratinocytes)
0.0

TNFalpha + IL-1beta

LAK cells IL-2
1.4
Liver cirrhosis
3.9

LAK cells IL-2 + IL-12
0.0
Lupus kidney
1.0

LAK cells IL-2 + IFN
1.2
NCI-H292 none
0.0

gamma

LAK cells IL-2 + IL-18
1.1
NCI-H292 IL-4
0.0

LAK cells
0.0
NCI-H292 IL-9
0.0

PMA/ionomycin

NK Cells IL-2 rest
0.0
NCI-H292 IL-13
1.2

Two Way MLR 3 day
2.4
NCI-H292 IFN gamma
1.5

Two Way MLR 5 day
0.0
HPAEC none
0.0

Two Way MLR 7 day
0.0
HPAEC TNFalpha + IL-1beta
0.0

PBMC rest
0.0
Lung fibroblast none
1.5

PBMC PWM
1.2
Lung fibroblast TNFalpha + IL-
1.2

1beta

PBMC PHA-L
0.0
Lung fibroblast IL-4
0.0

Ramos (B cell) none
34.4
Lung fibroblast IL-9
0.0

Ramos (B cell) ionomycin
100.0
Lung fibroblast IL-13
0.0

B lymphocytes PWM
1.2
Lung fibroblast IFN gamma
0.0

B lymphocytes CD40L
0.8
Dermal fibroblast CCD1070 rest
0.0

and IL-4

EOL-1 dbcAMP
0.0
Dermal fibroblast CCD1070
1.0

TNF alpha

EOL-1 dbcAMP
0.0
Dermal fibroblast CCD1070 IL-
0.0

PMA/ionomycin

1beta

Dendritic cells none
0.0
Dermal fibroblast IFN gamma
2.1

Dendritic cells LPS
1.2
Dermal fibroblast IL-4
0.0

Dendritic cells anti-CD40
0.0
IBD Colitis 2
1.4

Monocytes rest
0.0
IBD Crohn's
1.0

Monocytes LPS
0.0
Colon
1.2

Macrophages rest
0.0
Lung
0.0

Macrophages LPS
0.0
Thymus
6.8

HUVEC none
2.1
Kidney
5.4

HUVEC starved
0.0

[0553] Panel 1.3D Summary: Ag2618 The CG56720-01 gene shows highest expression in a sample derived from a colon cancer cell line (HCT116)(CT=28.3). In addition, there is substantial expression in other colon cancer cell lines (SW620), melanoma cell lines (SK-MEL-5, LOXIMVI) and lung cancer cell lines (LX-1, SHP-77). Thus, this gene could be used to distinguish these cell lines from other samples in the panel. Moreover, therapeutic modulation of this gene, through the use of small molecule drugs, antibodies or protein therapeutics might be of use in the treatment of melanoma, lung cancer or colon cancer. In addition, this gene is expressed at much higher levels in the SW620 colon cancer cell line, than in the SW480 cell line, where the SW620 sample is derived from a metastasis of the SW480 primary tumor. Thus, expression of this gene could be used to differentiate between the two forms of colon cancer.

[0554] Panel 2.2 Summary: Ag2618 The expression of the CG56720-01 gene is highest in a sample derived from a sample of normal kidney adjacent to a kidney malignancy. (CT=33.2) In addition, there is expression associated with an ovarian cancer. Thus, the expression of this gene could be used to distinguish these two samples from the other samples in the panel. Moreover, therapeutic modulation of this gene, through the use of small molecule drugs, antibodies or protein therapeutics might be of benefit in the treatment of ovarian cancer.

[0555] Panel 4D Summary: Ag2618 The highest expression of CG56720-01 is detected in Ramos B cells stimulated with ionomycin (CT=30.1). Lower but still significant levels of expression are seen in untreated Ramos B cells (CT=31.6), normal thymus and kidney tissue samples. B cells represent a principle component of immunity and contribute to the immune response in a number of important functional roles, including antibody production. For example, production of antibodies against self-antigens is a major component in autoimmune disorders. Since these cells play an important role in inflammatory processes and inflammatory cascades, therapeutic modulation of this gene product may reduce or eliminate the symptoms of patients suffering from asthma, allergies, chronic obstructive pulmonary disease, emphysema, Crohn's disease, ulcerative colitis, rheumatoid arthritis, psoriasis, osteoarthritis, and other autoimmune disorders.

[0556] Q. CG148698-02/GMAC026083_C: Olfactory Receptor

[0557] Expression of gene CG148698-02 was assessed using the primer-probe sets Ag1533, Ag2617 and Ag2862, described in Tables QA, QB and QC. Results of the RTQ-PCR runs are shown in Tables QD, QE, QF, QG, QH, QI and QJ.

72TABLE QAProbe Name Ag1533StartSEQ IDPrimersSequencesLengthPositionNO:Forward5′-accatcatcaagagtgctatgg-3′22446235ProbeTET-5′-tcctttcgaagcttctgcatcatcct-3′-TAMRA26473236Reverse5′-aggcatgtcagcaagaatacat-3′22504237

[0558]

73

TABLE QB

Probe Name Ag2617

Start
SEQ ID

Primers
Sequences
Length
Position
NO:

Forward
5′-ccatggcatttgatcactatgt-3′
22
378
238

Probe
TET-5′-tgagatataccaccatcttgactccca-3′-TAMRA
27
417
239

Reverse
5′-ccatagcactcttgatgatggt-3′
22
446
240

[0559]

74

TABLE QC

Probe Name Ag2862

Start
SEQ ID

Primers
Sequences
Lenght
Position
NO:

Forward
5′-ccatggcatttgatcactatgt-3′
22
378
241

Probe
TET-5′-tgagatataccaccatcttgactccca-3′-TAMRA
27
417
242

Reverse
5′-ccatagcactcttgatgatggt-3′
22
446
243

[0560]

75

TABLE QD

CNS_neurodegeneration_v1.0

Rel. Exp. (%)

Rel. Exp. (%)

Ag1533, Run

Ag1533, Run

Tissue Name
225432469
Tissue Name
225432469

AD 1 Hippo
36.6
Control (Path) 3
16.5

Temporal Ctx

AD 2 Hippo
11.6
Control (Path) 4
42.9

Temporal Ctx

AD 3 Hippo
50.7
AD 1 Occipital Ctx
39.2

AD 4 Hippo
75.3
AD 2 Occipital Ctx (Missing)
0.0

AD 5 Hippo
20.4
AD 3 Occipital Ctx
25.9

AD 6 Hippo
69.3
AD 4 Occipital Ctx
82.9

Control 2 Hippo
22.5
AD 5 Occipital Ctx
11.0

Control 4 Hippo
53.2
AD 5 Occipital Ctx
7.1

Control (Path) 3 Hippo
29.7
Control 1 Occipital Ctx
11.9

AD 1 Temporal Ctx
96.6
Control 2 Occipital Ctx
29.1

AD 2 Temporal Ctx
34.6
Control 3 Occipital Ctx
41.2

AD 3 Temporal Ctx
54.3
Control 4 Occipital Ctx
11.9

AD 4 Temporal Ctx
68.3
Control (Path) 1
23.3

Occipital Ctx

AD 5 Inf Temporal Ctx
90.8
Control (Path) 2
14.3

Occipital Ctx

AD 5 Sup Temporal
53.2
Control (Path) 3
35.1

Ctx

Occipital Ctx

AD 6 Inf Temporal Ctx
62.0
Control (Path) 4
30.8

Occipital Ctx

AD 6 Sup Temporal
55.5
Control 1 Parietal Ctx
18.2

Ctx

Control 1 Temporal Ctx
5.3
Control 2 Parietal Ctx
69.3

Control 2 Temporal Ctx
19.6
Control 3 Parietal Ctx
6.3

Control 3 Temporal Ctx
38.7
Control (Path) 1
33.9

Parietal Ctx

Control 3 Temporal Ctx
17.0
Control (Path) 2
18.4

Parietal Ctx

Control (Path) 1
29.1
Control (Path) 3
26.8

Temporal Ctx

Parietal Ctx

Control (Path) 2
23.7
Control (Path) 4
100.0

Temporal Ctx

Parietal Ctx

[0561]

76

TABLE QE

General_screening_panel_v1.5

Rel. Exp. (%)

Rel. Exp. (%)

Ag1533, Run

Ag1533, Run

Tissue Name
228632846
Tissue Name
228632846

Adipose
12.9
Renal ca. TK-10
2.4

Melanoma* Hs688(A).T
1.5
Bladder
43.8

Melanoma* Hs688(B).T
0.4
Gastric ca. (liver met.)
54.7

NCI-N87

Melanoma* M14
0.0
Gastric ca. KATO III
0.4

Melanoma* LOXIMVI
0.5
Colon ca. SW-948
0.0

Melanoma* SK-MEL-5
0.3
Colon ca. SW480
0.0

Squamous cell
0.0
Colon ca.* (SW480 met)
0.0

carcinoma SCC-4

SW620

Testis Pool
15.1
Colon ca. HT29
0.0

Prostate ca.* (bone met)
1.9
Colon ca. HCT-116
2.8

PC-3

Prostate Pool
23.7
Colon ca. CaCo-2
0.7

Placenta
2.9
Colon cancer tissue
1.5

Uterus Pool
11.5
Colon ca. SW1116
0.3

Ovarian ca. OVCAR-3
25.5
Colon ca. Colo-205
0.0

Ovarian ca. SK-OV-3
87.7
colon ca. SW-48
0.0

Ovarian ca. OVCAR-4
0.0
Colon Pool
37.4

Ovarian ca. OVCAR-5
10.1
Small Intestine Pool
43.5

Ovarian ca. IGROV-1
0.0
Stomach Pool
27.4

Ovarian ca. OVCAR-8
2.4
Bone Marrow Pool
22.1

Ovary
22.2
Fetal Heart
57.0

Breast ca. MCF-7
0.0
Heart Pool
12.2

Breast ca. MDA-MB-
1.3
Lymph Node Pool
66.9

231

Breast ca. BT 549
1.4
Fetal Skeletal Muscle
11.4

Breast ca. T47D
0.8
Skeletal Muscle Pool
7.5

Breast ca. MDA-N
0.7
Spleen Pool
21.3

Breast Pool
59.5
Thymus Pool
38.7

Trachea
19.1
CNS cancer (glio/astro)
1.6

U87-MG

Lung
27.2
CNS cancer (glio/astro) U-
1.5

118-MG

Fetal Lung
100.0
CNS cancer (neuro;met)
0.0

SK-N-AS

Lung ca. NCI-N417
0.0
CNS cancer (astro) SF-539
1.1

Lung ca. LX-1
0.4
CNS cancer (astro) SNB-75
4.5

Lung ca. NCI-H146
4.7
CNS cancer (glio) SNB-19
1.1

Lung ca. SHP-77
0.0
CNS cancer (glio) SF-295
20.6

Lung ca. A549
2.9
Brain (Amygdala) Pool
2.1

Lung ca. NCI-H526
0.0
Brain (cerebellum)
0.9

Lung ca. NCI-H23
6.7
Brain (fetal)
7.9

Lung ca. NCI-H460
9.2
Brain (Hippocampus) Pool
1.6

Lung ca. HOP-62
8.4
Cerebral Cortex Pool
2.6

Lung ca. NCI-H522
0.0
Brain (Substantia nigra)
2.0

Pool

Liver
0.4
Brain (Thalamus) Pool
2.5

Fetal Liver
8.7
Brain (whole)
3.8

Liver ca. HepG2
0.7
Spinal Cord Pool
5.0

Kidney Pool
67.8
Adrenal Gland
11.0

Fetal Kidney
94.6
Pituitary gland Pool
5.3

Renal ca. 786-0
1.2
Salivary Gland
1.7

Renal ca. A498
1.8
Thyroid (female)
2.5

Renal ca. ACHN
3.6
Pancreatic ca. CAPAN2
4.7

Renal ca. UO-31
5.8
Pancreas Pool
39.8

[0562]

77

TABLE QF

Panel 1.2

Rel. Exp. (%)

Rel. Exp. (%)

Ag1533, Run

Ag1533, Run

Tissue Name
142223910
Tissue Name
142223910

Endothelial cells
4.4
Renal ca. 786-0
1.8

Heart (Fetal)
3.9
Renal ca. A498
7.5

Pancreas
4.8
Renal ca. RXF 393
1.8

Pancreatic ca. CAPAN 2
1.1
Renal ca. ACHN
5.0

Adrenal Gland
20.9
Renal ca. UO-31
9.0

Thyroid
1.1
Renal ca. TK-10
2.3

Salivary gland
52.1
Liver
9.8

Pituitary gland
0.7
Liver (fetal)
6.0

Brain (fetal)
0.7
Liver ca. (hepatoblast)
1.0

HepG2

Brain (whole)
0.0
Lung
1.5

Brain (amygdala)
1.0
Lung (fetal)
2.9

Brain (cerebellum)
0.3
Lung ca. (small cell) LX-1
0.2

Brain (hippocampus)
6.3
Lung ca. (small cell)
5.2

NCI-H69

Brain (thalamus)
1.9
Lung ca. (s.cell var.)
0.0

SHP-77

Cerebral Cortex
9.2
Lung ca. (large cell)NCI-
2.8

H460

Spinal cord
2.9
Lung ca. (non-sm. cell)
5.1

A549

glio/astro U87-MG
1.6
Lung ca. (non-s.cell)
10.2

NCI-H23

glio/astro U-118-MG
1.1
Lung ca. (non-s.cell)
28.3

HOP-62

astrocytoma SW 1783
1.2
Lung ca. (non-s.cl) NCI-
2.0

H522

neuro*; met SK-N-AS
0.2
Lung ca. (squam.) SW
3.6

900

astrocytoma SF-539
7.4
Lung ca. (squam.) NCI-
0.7

H596

astrocytoma SNB-75
1.6
Mammary gland
3.4

glioma SNB-19
8.1
Breast ca.* (pl.ef) MCF-7
1.3

glioma U251
5.9
Breast ca.* (pl.ef) MDA-
0.3

MB-231

glioma SF-295
12.2
Breast ca.* (pl. ef) T47D
12.8

Heart
15.3
Breast ca. BT-549
2.9

Skeletal Muscle
4.9
Breast ca. MDA-N
0.8

Bone marrow
4.4
Ovary
9.3

Thymus
0.8
Ovarian ca. OVCAR-3
42.0

Spleen
5.6
Ovarian ca. OVCAR-4
1.4

Lymph node
1.6
Ovarian ca. OVCAR-5
18.2

Colorectal
10.9
Ovarian ca. OVCAR-8
0.0

Stomach
3.2
Ovarian ca. IGROV-1
0.0

Small intestine
7.8
Ovarian ca. (ascites) SK-
100.0

OV-3

Colon ca. SW480
0.0
Uterus
6.5

Colon ca.* SW620
0.0
Placenta
1.8

(SW480 met)

Colon ca. HT29
0.9
Prostate
23.7

Colon ca. HCT-116
1.8
Prostate ca.* (bone met)
4.5

PC-3

Colon ca. CaCo-2
1.5
Testis
2.7

CC Well to Mod Diff
2.9
Melanoma Hs688(A).T
0.2

(ODO3866)

Colon ca. HCC-2998
11.6
Melanoma* (met)
0.9

Hs688(B).T

Gastric ca. (liver met)
44.4
Melanoma UACC-62
4.5

NCI-N87

Bladder
98.6
Melanoma M14
4.2

Trachea
1.5
Melanoma LOX IMVI
0.4

Kidney
40.1
Melanoma* (met) SK-
0.0

MEL-5

Kidney (fetal)
25.5

[0563]

78

TABLE QG

Panel 1.3D

Rel. Exp. (%)

Rel. Exp. (%)

Ag2617, Run

Ag2617, Run

Tissue Name
167644078
Tissue Name
167644078

Liver adenocarcinoma
2.3
Kidney (fetal)
15.0

Pancreas
2.2
Renal ca. 786-0
1.5

Pancreatic ca. CAPAN 2
0.0
Renal ca. A498
3.1

Adrenal gland
0.0
Renal ca. RXF 393
0.0

Thyroid
1.5
Renal ca. ACHN
2.2

Salivary gland
3.2
Renal ca. UO-31
0.0

Pituitary gland
6.1
Renal ca. TK-10
2.4

Brain (fetal)
2.9
Liver
2.5

Brain (whole)
0.0
Liver (fetal)
0.0

Brain (amygdala)
0.0
Liver ca. (hepatoblast)
0.0

HepG2

Brain (cerebellum)
0.0
Lung
2.6

Brain (hippocampus)
3.2
Lung (fetal)
4.0

Brain (substantia nigra)
2.3
Lung ca. (small cell) LX-1
0.0

Brain (thalamus)
0.0
Lung ca. (small cell)
0.0

NCI-H69

Cerebral Cortex
1.3
Lung ca. (s.cell var.)
0.0

SHP-77

Spinal cord
3.4
Lung ca. (large cell)NCI-
0.0

H460

glio/astro U87-MG
0.0
Lung ca. (non-sm. cell)
0.0

A549

glio/astro U-118-MG
1.5
Lung ca. (non-s.cell)
4.7

NCI-H23

astrocytoma SW1783
2.0
Lung ca. (non-s.cell)
2.1

HOP-62

neuro*; met SK-N-AS
0.0
Lung ca. (non-s.cl) NCI-
0.0

H522

astrocytoma SF-539
6.3
Lung ca. (squam.) SW
3.0

900

astrocytoma SNB-75
1.4
Lung ca. (squam.) NCI-
0.0

H596

glioma SNB-19
2.5
Mammary gland
3.3

glioma U251
16.6
Breast ca.* (pl.ef) MCF-7
0.0

glioma SF-295
12.5
Breast ca.* (pl.ef) MDA-
0.0

MB-231

Heart (Fetal)
0.0
Breast ca.* (pl. ef) T47D
1.4

Heart
2.1
Breast ca. BT-549
1.3

Skeletal muscle (Fetal)
3.1
Breast ca. MDA-N
1.7

Skeletal muscle
0.0
Ovary
0.0

Bone marrow
1.9
Ovarian ca. OVCAR-3
9.2

Thymus
4.1
Ovarian ca. OVCAR-4
0.0

Spleen
1.6
Ovarian ca. OVCAR-5
9.2

Lymph node
7.1
Ovarian ca. OVCAR-8
5.1

Colorectal
3.8
Ovarian ca. IGROV-1
0.0

Stomach
0.0
Ovarian ca. (ascites) SK-
100.0

OV-3

Small intestine
1.4
Uterus
5.1

Colon ca. SW480
1.5
Placenta
0.0

Colon ca.* SW620
0.0
Prostate
4.5

(SW480 met)

Colon ca. HT29
1.0
Prostate ca.* (bone met)
1.0

PC-3

Colon ca. HCT-116
0.0
Testis
5.4

Colon ca. CaCo-2
0.0
Melanoma Hs688(A).T
0.0

CC Well to Mod Diff
0.0
Melanoma* (met)
0.9

(ODO3866)

Hs688(B).T

Colon ca. HCC-2998
0.0
Melanoma UACC-62
0.0

Gastric ca. (liver met)
12.7
Melanoma M14
0.0

NCI-N87

Bladder
16.4
Melanoma LOX IMVI
0.0

Trachea
4.1
Melanoma* (met) SK-
0.0

MEL-5

Kidney
0.0
Adipose
12.3

[0564]

79

TABLE QH

Panel 2D

Rel. Exp. (%)

Rel. Exp. (%)

Ag1533, Run

Ag1533, Run

Tissue Name
145165498
Tissue Name
145165498

Normal Colon
48.0
Kidney Margin 8120608
6.1

CC Well to Mod Diff
5.6
Kidney Cancer 8120613
0.0

(ODO3866)

CC Margin (ODO3866)
4.6
Kidney Margin 8120614
6.9

CC Gr.2 rectosigmoid
3.5
Kidney Cancer 9010320
10.2

(ODO3868)

CC Margin (ODO3868)
1.0
Kidney Margin 9010321
15.7

CC Mod Diff (ODO3920)
4.4
Normal Uterus
36.3

CC Margin (ODO3920)
6.6
Uterine Cancer 064011
45.7

CC Gr.2 ascend colon
1.6
Normal Thyroid
8.1

(ODO3921)

CC Margin (ODO3921)
6.1
Thyroid Cancer
7.7

CC from Partial Hepatectomy
0.0
Thyroid Cancer A302152
27.0

(ODO4309) Mets

Liver Margin (ODO4309)
4.8
Thyroid Margin A302153
29.5

Colon mets to lung (OD04451-
4.3
Normal Breast
40.9

01)

Lung Margin (OD04451-02)
1.2
Breast Cancer
3.5

Normal Prostate 6546-1
26.1
Breast Cancer (OD04590-
18.9

01)

Prostate Cancer (OD04410)
64.2
Breast Cancer Mets
34.6

(OD04590-03)

Prostate Margin (OD04410)
43.8
Breast Cancer Metastasis
19.3

Prostate Cancer (OD04720-01)
42.0
Breast Cancer
21.2

Prostate Margin (OD04720-02)
34.2
Breast Cancer
35.1

Normal Lung
31.9
Breast Cancer 9100266
7.7

Lung Met to Muscle
0.0
Breast Margin 9100265
2.9

(ODO4286)

Muscle Margin (ODO4286)
1.9
Breast Cancer A209073
14.8

Lung Malignant Cancer
13.1
Breast Margin A2090734
23.3

(OD03126)

Lung Margin (OD03126)
26.8
Normal Liver
21.9

Lung Cancer (OD04404)
5.4
Liver Cancer
22.4

Lung Margin (OD04404)
37.9
Liver Cancer 1025
0.0

Lung Cancer (OD04565)
2.9
Liver Cancer 1026
2.1

Lung Margin (OD04565)
9.3
Liver Cancer 6004-T
13.5

Lung Cancer (OD04237-01)
24.8
Liver Tissue 6004-N
5.9

Lung Margin (OD04237-02)
10.5
Liver Cancer 6005-T
0.0

Ocular Mel Met to Liver
10.1
Liver Tissue 6005-N
0.0

(ODO4310)

Liver Margin (ODO4310)
8.7
Normal Bladder
42.0

Melanoma Metastasis
0.0
Bladder Cancer
5.8

Lung Margin (OD04321)
25.0
Bladder Cancer
65.1

Normal Kidney
100.0
Bladder Cancer
5.6

(OD04718-01)

Kidney Ca, Nuclear grade 2
52.1
Bladder Normal Adjacent
32.8

(OD04338)

(OD04718-03)

Kidney Margin (OD04338)
22.1
Normal Ovary
5.7

Kidney Ca Nuclear grade 1/2
41.5
Ovarian Cancer
9.4

(OD04339)

Kidney Margin (OD04339)
46.0
Ovarian Cancer
8.8

(OD04768-07)

Kidney Ca, Clear cell type
27.0
Ovary Margin (OD04768-
3.3

(OD04340)

08)

Kidney Margin (OD04340)
24.7
Normal Stomach
30.6

Kidney Ca, Nuclear grade 3
9.1
Gastric Cancer 9060358
0.0

(OD04348)

Kidney Margin (OD04348)
92.7
Stomach Margin 9060359
0.0

Kidney Cancer (OD04622-01)
5.0
Gastric Cancer 9060395
5.3

Kidney Margin (OD04622-03)
2.0
Stomach Margin 9060394
3.1

Kidney Cancer (OD04450-01)
10.9
Gastric Cancer 9060397
4.7

Kidney Margin (OD04450-03)
17.6
Stomach Margin 9060396
0.0

Kidney Cancer 8120607
0.8
Gastric Cancer 064005
11.3

[0565]

80

TABLE QI

Panel 4.1D

Rel. Exp. (%)

Rel. Exp. (%)

Ag1533, Run

Ag1533, Run

Tissue Name
223794696
Tissue Name
223794696

Secondary Th1 act
0.0
HUVEC IL-1beta
11.8

Secondary Th2 act
20.4
HUVEC IFN gamma
11.5

Secondary Tr1 act
13.1
HUVEC TNF alpha + IFN
4.0

gamma

Secondary Th1 rest
25.3
HUVEC TNF alpha + IL4
18.7

Secondary Th2 rest
8.0
HUVEC IL-11
29.1

Secondary Tr1 rest
33.9
Lung Microvascular EC none
83.5

Primary Th1 act
0.0
Lung Microvascular EC
14.0

TNF alpha + IL-1beta

Primary Th2 act
25.0
Microvascular Dermal EC none
8.5

Primary Tr1 act
7.9
Microsvasular Dermal EC
0.0

TNF alpha + IL-1beta

Primary Th1 rest
14.5
Bronchial epithelium TNF
15.2

alpha + IL1beta

Primary Th2 rest
4.3
Small airway epithelium none
0.0

Primary Tr1 rest
20.7
Small airway epithelium
20.9

TNF alpha + IL-1beta

CD45RA CD4 lymphocyte
21.2
Coronery artery SMC rest
0.0

act

CD45RO CD4 lymphocyte
65.5
Coronery artery SMC TNF
0.0

act

alpha + IL-1beta

CD8 lymphocyte act
25.9
Astrocytes rest
20.3

Secondary CD8
19.6
Astrocytes TNF alpha +
10.3

lymphocyte rest

IL-1beta

Secondary CD8
4.5
KU-812 (Basophil) rest
10.9

lymphocyte act

CD4 lymphocyte none
53.2
KU-812 (Basophil)
34.2

PMA/ionomycin

2ry Th1/Th2/Tr1_anti-
19.2
CCD1106 (Keratinocytes) none
0.0

CD95 CH11

LAK cells rest
22.1
CCD1106 (Keratinocytes)
8.5

TNF alpha + IL-1beta

LAK cells IL-2
51.1
Liver cirrhosis
26.6

LAK cells IL-2 + IL-12
3.1
NCI-H292 none
19.9

LAK cells IL-2 + IFN
24.0
NCI-H292 IL-4
27.5

gamma

LAK cells IL-2 + IL-18
14.6
NCI-H292 IL-9
29.3

LAK cells
0.0
NCI-H292 IL-13
16.2

PMA/ionomycin

NK Cells IL-2 rest
38.7
NCI-H292 IFN gamma
44.1

Two Way MLR 3 day
61.6
HPAEC none
8.5

Two Way MLR 5 day
35.6
HPAEC TNF alpha + IL-1beta
10.7

Two Way MLR 7 day
9.9
Lung fibroblast none
60.7

PBMC rest
26.2
Lung fibroblast TNF alpha + IL-
40.6

1beta

PBMC PWM
3.6
Lung fibroblast IL-4
8.8

PBMC PHA-L
4.5
Lung fibroblast IL-9
21.3

Ramos (B cell) none
0.0
Lung fibroblast IL-13
4.8

Ramos (B cell) ionomycin
0.0
Lung fibroblast IFN gamma
4.6

B lymphocytes PWM
5.1
Dermal fibroblast CCD1070 rest
0.0

B lymphocytes CD40L
17.0
Dermal fibroblast CCD1070
20.0

and IL-4

TNF alpha

EOL-1 dbcAMP
10.9
Dermal fibroblast CCD1070 IL-
4.0

1beta

EOL-1 dbcAMP
0.0
Dermal fibroblast IFN gamma
10.7

PMA/ionomycin

Dendritic cells none
4.1
Dermal fibroblast IL-4
26.6

Dendritic cells LPS
17.7
Dermal Fibroblasts rest
10.4

Dendritic cells-CD40
14.8
Neutrophils TNFa + LPS
10.0

Monocytes rest
10.7
Neutrophils rest
21.3

Monocytes LPS
16.4
Colon
0.0

Macrophages rest
18.6
Lung
4.2

Macrophages LPS
8.0
Thymus
100.0

HUVEC none
5.4
Kidney
81.8

HUVEC starved
2.6

[0566]

81

TABLE QJ

Panel 4D

Rel. Exp. (%)

Rel. Exp. (%)

Ag2862, Run

Ag2862, Run

Tissue Name
164299494
Tissue Name
164299494

Secondary Th1 act
0.0
HUVEC IL-1beta
0.0

Secondary Th2 act
10.9
HUVEC IFN gamma
4.2

Secondary Tr1 act
2.6
HUVEC TNF alpha + IFN
2.5

gamma

Secondary Th1 rest
4.5
HUVEC TNF alpha + IL4
4.6

Secondary Th2 rest
13.8
HUVEC IL-11
2.4

Secondary Tr1 rest
9.8
Lung Microvascular EC none
28.9

Primary Th1 act
0.0
Lung Microvascular EC
13.2

TNF alpha + IL-1beta

Primary Th2 act
0.0
Microvascular Dermal EC none
11.7

Primary Tr1 act
0.0
Microsvasular Dermal EC
4.3

TNF alpha + IL-1beta

Primary Th1 rest
65.5
Bronchial epithelium TNF
9.7

alpha + IL1beta

Primary Th2 rest
24.0
Small airway epithelium none
1.7

Primary Tr1 rest
6.8
Small airway epithelium
36.1

TNF alpha + IL-1beta

CD45RA CD4 lymphocyte
12.9
Coronery artery SMC rest
11.1

act

CD45RO CD4 lymphocyte
7.4
Coronery artery SMC TNF
6.7

act

alpha + IL-1beta

CD8 lymphocyte act
0.0
Astrocytes rest
9.4

Secondary CD8
13.7
Astrocytes TNF alpha +
8.0

lymphocyte rest

IL-1beta

Secondary CD8
0.0
KU-812 (Basophil) rest
5.9

lymphocyte act

CD4 lymphocyte none
22.1
KU-812 (Basophil)
25.9

PMA/ionomycin

2ry Th1/Th2/Tr1_anti-
20.2
CCD1106 (Keratinocytes) none
6.5

CD95 CH11

LAK cells rest
20.7
CCD1106 (Keratinocytes)
2.9

TNF alpha + IL-1beta

LAK cells IL-2
53.2
Liver cirrhosis
17.8

LAK cells IL-2 + IL-12
27.4
Lupus kidney
13.9

LAK cells IL-2 + IFN
73.2
NCI-H292 none
23.2

gamma

LAK cells IL-2 + IL-18
12.6
NCI-H292 IL-4
28.3

LAK cells
0.0
NCI-H292 IL-9
24.1

PMA/ionomycin

NK Cells IL-2 rest
26.8
NCI-H292 IL-13
2.3

Two Way MLR 3 day
82.9
NCI-H292 IFN gamma
24.1

Two Way MLR 5 day
12.1
HPAEC none
8.0

Two Way MLR 7 day
0.0
HPAEC TNF alpha + IL-1beta
6.3

PBMC rest
8.9
Lung fibroblast none
9.9

PBMC PWM
17.4
Lung fibroblast TNF alpha + IL-
18.8

1beta

PBMC PHA-L
7.0
Lung fibroblast IL-4
11.7

Ramos (B cell) none
0.0
Lung fibroblast IL-9
18.7

Ramos (B cell) ionomycin
4.5
Lung fibroblast IL-13
20.0

B lymphocytes PWM
2.5
Lung fibroblast IFN gamma
17.3

B lymphocytes CD40L
12.9
Dermal fibroblast CCD1070 rest
3.0

and IL-4

EOL-1 dbcAMP
4.7
Dermal fibroblast CCD1070
9.9

TNF alpha

EOL-1 dbcAMP
0.0
Dermal fibroblast CCD1070 IL-
5.1

PMA/ionomycin

1beta

Dendritic cells none
1.8
Dermal fibroblast IFN gamma
1.7

Dendritic cells LPS
2.8
Dermal fibroblast IL-4
24.5

Dendritic cells anti-CD40
0.0
IBD Colitis 2
1.8

Monocytes rest
6.0
IBD Crohn's
2.1

Monocytes LPS
0.0
Colon
14.9

Macrophages rest
0.0
Lung
11.7

Macrophages LPS
11.9
Thymus
82.9

HUVEC none
0.0
Kidney
100.0

HUVEC starved
3.1

[0567] CNS_neurodegeneration_v1.0 Summary: Ag1533 The CG148698-02 gene shows widespread expression across all regions of the brain, with highest expression in the parietal cortex of a control patient (CT=33.5). This gene appears to be upregulated in the temporal cortex of patients with Alzheimer's disease. The temporal cortex is a region that shows severe degeneration in Alzheimer's disease, suggesting the expression of this gene may play a role in the pathogenesis of this disease. Therapeutic modulation of this gene or treatment with an antagonist to the receptor may be of benefit in treating Alzheimer's disease or dementia.

[0568] Ag2862 Expression is low/undetected in all samples in this panel (CT>35). (Data not shown.)

[0569] General_screening_panel_v1.5 Summary: Ag1533 Highest expression of the CG148698-02 gene is in the fetal lung (CT=30). Significant levels of expression are also detected in adult lung. This expression profile suggests that the gene product may be involved in the normal homeostasis of the lung. Therefore, therapeutic modulation of the expression or function of this gene may be effective in the treatment of diseases of that affect the lung including asthma, emphysema, and acute respiratory distress syndrome (ARDS).

[0570] This gene is also moderately expressed in a variety of metabolic tissues including adipose, adult and fetal heart, adult and fetal skeletal muscle, adrenal, pituitary, thyroid and pancreas. Thus, this gene product may be a small molecule drug target for the treatment of metabolic disease, including obesity and Types 1 and 2 diabetes. Furthermore, this gene is differentially expressed in adult (CT value=37) versus fetal liver (CT values=33.5), and may be used to differentiate between the adult and fetal phenotype in this tissue.

[0571] There is moderate expression in some tissues of the central nervous system, including the fetal brain and the spinal cord. Please see CNS_neurodegeneration_vl.0 Summary for discussion of potential utility in the central nervous system.

[0572] Overall, the expression of this gene is largely associated with normal tissues. However, significant expression of this gene is seen in cell lines derived ovarian and gastric cancers.

[0573] Thus therapeutic modulation of this gene, through the use of small molecule drugs, antibodies or protein therapeutics might be of use in the treatment of ovarian or gastric cancer.

[0574] Panel 1.2 Summary: Ag1533 The expression of the CG148698-02 gene is highest in a sample derived from an ovarian cancer cell line (CT=29. l). This particular cell line was derived from a unique form of ovarian cancer, that being ascites. In addition, there appears to be substantial expression of this gene in samples derived from other ovarian cancer cell lines as well as normal bladder tissue, normal kidney tissue and a cell lined derived from a gastric cancer. Thus, the expression of this gene in these tissues could be used to distinguish these samples from other samples in the panel. Additionally, the expression of this gene could be used to distinguish ascites derived samples from other samples in the panel. Furthermore, therapeutic modulation of this gene, through the use of small molecule drugs, antibodies or protein therapeutics might be of benefit in the treatment of ovarian cancer.

[0575] This gene is also expressed in a variety of metabolic tissues including Ag1533 is modestly expressed (CT values=31-34) in a variety of metabolic tissues including adult and fetal liver, adult and fetal heart, skeletal muscle, adrenal and pancreas. As is seen from General_screening_panel_v0.5, this suggests a role of the gene product in metabolic function. Thus, this gene product may be a small molecule drug target for the treatment of metabolic disease, including obesity and Types 1 and 2 diabetes.

[0576] Panel 1.3D Summary: Ag2617 Expression of the CG148698-02 gene is exclusive to an ovarian cancer cell line (SK-OV-3)(CT=33.1). Expression in this cell line is also detected in Panel 1.2. Interestingly, this cell line was derived from a unique form of ovarian cancer, that being ascites. Thus, the expression of this gene could be used to distinguish samples derived from this cell line from other samples in the panel in addition to distinguishing ascites from other samples in the panel. Moreover, therapeutic modulation of this gene, through the use of small molecule drugs, antibodies or protein therapeutics might be of benefit in the treatment of ovarian cancer.

[0577] Panel 2D Summary: Ag1533 The expression of the CG148698-02 gene appears to be highest in a sample derived from normal kidney tissue (CT=31.9). In addition there is substantial expression in samples derived from other samples of normal kidney tissue adjacent to malignant kidney. Moreover, there also appears to be expression associated with tissues, normal or malignant, derived from uterus, prostate, breast, bladder and thyroid. Thus, the expression of this gene could be used to distinguish samples derived from these tissue types when compared to other samples in the panel. Further, therapeutic modulation of this gene, or gene product, through the use of small molecule drugs, antibodies or protein therapeutics might be of benefit in the treatment of cancers of the above listed tissues.

[0578] Panel 4.1D Summary: Ag1533 The CG148698-02 transcript is expressed on most tissues in panel 4.1 D regardless of treatment, with highest expression in the thymus(CT=32.8). This transcript encodes a GPCR-like molecule with potential signaling activity and may important in maintaining normal cellular functions in a number of tissues. Therapies designed with the protein encoded for by this transcript could be important in regulating cellular viability or function.

[0579] Panel 4D Summary: Ag2862 The CG148698-02 transcript appears to be expressed in this panel regardless of treatment, with highest expression in the kidney (CT=33.2). This result is concordant with the results from Panel 2D. This transcript encodes a GPCR-like molecule with potential signaling activity and may important in maintaining normal cellular functions in a number of tissues. Therapies designed with the protein encoded for by this transcript could be important in regulating cellular viability or function.

[0580] R. CG153463-01/GMAC026083_B: Olfactory Receptor

[0581] Expression of gene CG 153463-01 was assessed using the primer-probe set Ag2616, described in Table RA. Results of the RTQ-PCR runs are shown in Tables RB and RC.

82TABLE RAProbe Name Ag2616StartSEQ IDPrimersSequencesLengthPositionNO:Forward5′-tcatcttcttatggccaatgtc-3′22830244ProbeTET-5′-tgcctcccatgcttaacccaatcata-3′-TAMRA26862245Reverse5′-ggtggatctccttggtcttaat-3′22894246

[0582]

83

TABLE RB

Panel 1.3D

Rel. Exp. (%)

Rel. Exp. (%)

Ag2616, Run

Ag2616, Run

Tissue Name
167644789
Tissue Name
167644789

Liver adenocarcinoma
11.0
Kidney (fetal)
2.3

Pancreas
0.0
Renal ca. 786-0
0.0

Pancreatic ca. CAPAN 2
0.0
Renal ca. A498
0.0

Adrenal gland
0.0
Renal ca. RXF 393
0.0

Thyroid
0.0
Renal ca. ACHN
0.0

Salivary gland
0.0
Renal ca. UO-31
0.0

Pituitary gland
0.6
Renal ca. TK-10
0.0

Brain (fetal)
0.0
Liver
0.4

Brain (whole)
0.0
Liver (fetal)
0.0

Brain (amygdala)
0.0
Liver ca. (hepatoblast)
0.0

HepG2

Brain (cerebellum)
0.7
Lung
0.0

Brain (hippocampus)
0.3
Lung (fetal)
1.1

Brain (substantia nigra)
0.0
Lung ca. (small cell) LX-1
32.1

Brain (thalamus)
0.0
Lung ca. (small cell)
0.0

NCI-H69

Cerebral Cortex
0.3
Lung ca. (s.cell var.)
12.9

SHP-77

Spinal cord
0.4
Lung ca. (large cell)NCI-
0.0

H460

glio/astro U87-MG
0.0
Lung ca. (non-sm. cell)
0.6

A549

glio/astro U-118-MG
0.8
Lung ca. (non-s.cell)
0.0

NCI-H23

astrocytoma SW1783
0.4
Lung ca. (non-s.cell)
0.0

HOP-62

neuro*; met SK-N-AS
0.0
Lung ca. (non-s.cl) NCI-
0.0

H522

astrocytoma SF-539
1.2
Lung ca. (squam.) SW
0.0

900

astrocytoma SNB-75
0.0
Lung ca. (squam.) NCI-
0.0

H596

glioma SNB-19
0.0
Mammary gland
0.2

glioma U251
1.3
Breast ca.* (pl.ef) MCF-7
0.0

glioma SF-295
1.5
Breast ca.* (pl.ef) MDA-
0.0

MB-231

Heart (Fetal)
0.0
Breast ca.* (pl. ef) T47D
0.0

Heart
0.0
Breast ca. BT-549
0.0

Skeletal muscle (Fetal)
0.5
Breast ca. MDA-N
11.9

Skeletal muscle
0.0
Ovary
0.0

Bone marrow
0.0
Ovarian ca. OVCAR-3
1.8

Thymus
0.0
Ovarian ca. OVCAR-4
0.5

Spleen
0.6
Ovarian ca. OVCAR-5
0.8

Lymph node
0.0
Ovarian ca. OVCAR-8
0.0

Colorectal
0.9
Ovarian ca. IGROV-1
0.5

Stomach
0.6
Ovarian ca. (ascites) SK-
8.4

OV-3

Small intestine
0.0
Uterus
0.3

Colon ca. SW480
0.0
Placenta
0.5

Colon ca.* SW620
13.1
Prostate
0.0

(SW480 met)

Colon ca. HT29
0.0
Prostate ca.* (bone met)
0.0

PC-3

Colon ca. HCT-116
100.0
Testis
0.0

Colon ca. CaCo-2
0.0
Melanoma Hs688(A).T
0.0

CC Well to Mod Diff
0.0
Melanoma* (met)
0.0

(ODO3866)

Hs688(B).T

Colon ca. HCC-2998
0.5
Melanoma UACC-62
0.8

Gastric ca. (liver met)
0.0
Melanoma M14
0.8

NCI-N87

Bladder
0.8
Melanoma LOX IMVI
14.2

Trachea
0.0
Melanoma* (met) SK-
51.4

MEL-5

Kidney
1.1
Adipose
1.1

[0583]

84

TABLE RC

Panel 2.2

Rel. Exp. (%) Ag2616,

Rel. Exp. (%) Ag2616,

Tissue Name
Run 175063646
Tissue Name
Run 175063646

Normal Colon
5.4
Kidney Margin (OD04348)
100.0

Colon cancer (OD06064)
0.0
Kidney malignant cancer
0.0

(OD06204B)

Colon Margin (OD06064)
0.0
Kidney normal adjacent
0.0

tissue (OD06204E)

Colon cancer (OD06159)
0.0
Kidney Cancer (OD04450-
0.0

01)

Colon Margin (OD06159)
4.9
Kidney Margin (OD04450-
2.5

03)

Colon cancer (OD06297-04)
0.0
Kidney Cancer 8120613
0.0

Colon Margin (OD06297-
2.9
Kidney Margin 8120614
0.0

015)

CC Gr.2 ascend colon
0.0
Kidney Cancer 9010320
5.7

(ODO3921)

CC Margin (ODO3921)
0.0
Kidney Margin 9010321
0.0

Colon cancer metastasis
0.0
Kidney Cancer 8120607
0.0

(OD06104)

Lung Margin (OD06104)
0.0
Kidney Margin 8120608
0.0

Colon mets to lung
5.1
Normal Uterus
11.8

(OD04451-01)

Lung Margin (OD04451-02)
0.0
Uterine Cancer 064011
14.0

Normal Prostate
0.0
Normal Thyroid
0.0

Prostate Cancer (OD04410)
0.0
Thyroid Cancer
0.0

Prostate Margin (OD04410)
13.9
Thyroid Cancer A302152
6.6

Normal Ovary
14.1
Thyroid Margin A302153
4.4

Ovarian cancer (OD06283-
0.0
Normal Breast
0.0

03)

Ovarian Margin (OD06283-
22.1
Breast Cancer
8.4

07)

Ovarian Cancer
20.0
Breast Cancer
0.0

Ovarian cancer (OD06145)
0.0
Breast Cancer (OD04590-
5.6

01)

Ovarian Margin (OD06145)
5.6
Breast Cancer Mets
0.0

(OD04590-03)

Ovarian cancer (OD06455-
0.0
Breast Cancer Metastasis
0.0

03)

Ovarian Margin (OD06455-
0.0
Breast Cancer
0.0

07)

Normal Lung
7.7
Breast Cancer 9100266
0.0

Invasive poor diff. lung
59.9
Breast Margin 9100265
0.0

adeno (ODO4945-01

Lung Margin (ODO4945-03)
0.0
Breast Cancer A209073
0.0

Lung Malignant Cancer
0.0
Breast Margin A2090734
0.0

(OD03126)

Lung Margin (OD03126)
3.1
Breast cancer (OD06083)
22.4

Lung Cancer (OD05014A)
0.0
Breast cancer node
7.5

metastasis (OD06083)

Lung Margin (OD05014B)
2.5
Normal Liver
3.6

Lung cancer (OD06081)
0.0
Liver Cancer 1026
0.0

Lung Margin (OD06081)
0.0
Liver Cancer 1025
0.0

Lung Cancer (OD04237-01)
0.0
Liver Cancer 6004-T
0.0

Lung Margin (OD04237-02)
6.2
Liver Tissue 6004-N
5.0

Ocular Mel Met to Liver
0.0
Liver Cancer 6005-T
0.0

(ODO4310)

Liver Margin (OD04310)
7.9
Liver Tissue 6005-N
0.0

Melanoma Metastasis
0.0
Liver Cancer
7.2

Lung Margin (OD04321)
0.0
Normal Bladder
5.3

Normal Kidney
5.5
Bladder Cancer
0.0

Kidney Ca, Nuclear grade 2
12.8
Bladder Cancer
6.9

(OD04338)

Kidney Margin (OD04338)
0.8
Normal Stomach
11.6

Kidney Ca Nuclear grade 1/2
42.9
Gastric Cancer 9060397
0.0

(OD04339)

Kidney Margin (OD04339)
7.3
Stomach Margin 9060396
0.0

Kidney Ca, Clear cell type
0.0
Gastric Cancer 9060395
0.0

(OD04340)

Kidney Margin (OD04340)
26.1
Stomach Margin 9060394
0.0

Kidney Ca, Nuclear grade 3
0.0
Gastric Cancer 064005
0.0

(OD04348)

[0584] Panel 1.3D Summary: Ag2616 The expression of the CG153463-01 gene appears to be highest in a sample derived from a colon cancer cell lint (HCT116)(CT=29.3). In addition, there is expression evident in another colon cancer cell line (SW620), two melanoma cell lines (SK-MEL-5, LOX IMVI), two lung cancer cell lines (LX-1, SHP-77), and ovarian cancer cell line, a breast cancer cell line and a liver cancer. Thus, the expression of this gene could be used to distinguish HCT-116 cells for the other cells in the panel. Moreover, therapeutic modulation of this gene, through the use of antibodies, small molecule drugs or protein therapeutics might be of benefit in the treatment of colon cancer, melanoma, lung cancer, ovarian cancer, breast cancer or liver cancer.

[0585] Panel 2.2 Summary: Ag2616 The expression of the CG153463-01 gene is highest in a sample derived from normal kidney tissue adjacent to a malignancy (CT=32.9). In addition, there appears to be expression in another normal kidney tissue sample, a sample of malignant kidney and a sample from a lung cancer. Thus, the expression of this gene could be used to distinguish these samples from other samples in the panel. Moreover, therapeutic modulation of this gene, through the use of antibodies, small molecule drugs or protein therapeutics might be of benefit in the treatment of lung cancer or kidney cancer

[0586] Panel 4D Summary: Ag2616 Data from one experiment with this probe and primer is not included because the amp plot indicates that there were experimental difficulties. (Data not shown.)

[0587] S. CG56808-01/GMAC026083 A: Olfactory Receptor

[0588] Expression of gene CG56808-01 was assessed using the primer-probe set Ag2615, described in Table SA. Results of the RTQ-PCR runs are shown in Tables SB, SC and SD.

85TABLE SAProbe Name Ag2615StartSEQ IDPrimersSequencesLengthPositionNO:Forward5′-ctatgtgctcattctgcgttct-3′22662247ProbeTET-5′-actgcttcccgtgaggaacgcct-3′-TAMRA23693248Reverse5′-cacacatgtgttgagagctttg-3′22716249

[0589]

86

TABLE SB

Panel 1.3D

Rel. Exp. (%) Ag2615, Run

Rel. Exp. (%) Ag2615, Run

Tissue Name
167644077
Tissue Name
167644077

Liver adenocarcinoma
0.0
Kidney (fetal)
1.1

Pancreas
0.0
Renal ca. 786-0
0.0

Pancreatic ca. CAPAN 2
0.0
Renal ca. A498
0.0

Adrenal gland
0.6
Renal ca. RXF 393
0.2

Thyroid
0.4
Renal ca. ACHN
0.0

Salivary gland
0.6
Renal ca. UO-31
0.0

Pituitary gland
0.0
Renal ca. TK-10
0.0

Brain (fetal)
1.0
Liver
0.3

Brain (whole)
0.6
Liver (fetal)
0.0

Brain (amygdala)
0.0
Liver ca. (hepatoblast)
0.5

HepG2

Brain (cerebellum)
0.0
Lung
0.9

Brain (hippocampus)
0.0
Lung (fetal)
0.1

Brain (substantia nigra)
0.8
Lung ca. (small cell) LX-1
37.1

Brain (thalamus)
0.0
Lung ca. (small cell)
0.0

NCI-H69

Cerebral Cortex
0.6
Lung ca. (s. cell var.)
3.6

SHP-77

Spinal cord
0.4
Lung ca. (large cell) NCI-
0.0

H460

glio/astro U87-MG
0.0
Lung ca. (non-sm. cell)
0.7

A549

glio/astro U-118-MG
0.2
Lung ca. (non-s. cell)
0.0

NCI-H23

astrocytoma SW1783
0.0
Lung ca. (non-s. cell)
0.0

HOP-62

neuro*; met SK-N-AS
0.0
Lung ca. (non-s. cl) NCI-
0.0

H522

astrocytoma SF-539
0.2
Lung ca. (squam.) SW
0.0

900

asrocytoma SNB-75
0.0
Lung ca. (squam.) NCI-
0.0

H596

glioma SNB-19
0.2
Mammary gland
0.2

glioma U251
0.8
Breast ca.* (pl.ef) MCF-7
0.0

glioma SF-295
1.0
Breast ca.* (pl.ef) MDA-
0.0

MB-231

Heart (fetal)
0.0
Breast ca.* (pl.ef) T47D
0.0

Heart
0.6
Breast ca. BT-549
0.1

Skeletal muscle (fetal)
0.3
Breast ca. MDA-N
10.6

Skeletal muscle
0.0
Ovary
0.0

Bone marrow
0.2
Ovarian ca. OVCAR-3
0.7

Thymus
0.2
Ovarian ca. OVCAR-4
0.0

Spleen
0.3
Ovarian ca. OVCAR-5
0.5

Lymph node
0.2
Ovarian ca. OVCAR-8
0.0

Colorectal
0.1
Ovarian ca. IGROV-1
0.0

Stomach
0.0
Ovarian ca.* (ascites)
5.8

SK-OV-3

Small intestine
0.0
Uterus
0.6

Colon ca. SW480
0.4
Placenta
0.0

Colon ca.* SW620(SW480
9.9
Prostate
0.5

met)

Colon ca. HT29
0.0
Prostate ca.* (bone
0.0

met)PC-3

Colon ca. HCT-116
100.0
Testis
0.4

Colon ca. CaCo-2
0.2
Melanoma Hs688(A).T
0.0

Colon ca.
0.0
Melanoma* (met)
0.0

tissue(ODO3866)

Hs688(B).T

Colon ca. HCC-2998
0.4
Melanoma UACC-62
0.0

Gastric ca.* (liver met)
0.0
Melanoma M14
0.2

NCI-N87

Bladder
2.5
Melanoma LOX IMVI
9.4

Trachea
0.0
Melanoma* (met) SK-
36.3

MEL-5

Kidney
1.5
Adipose
1.2

[0590]

87

TABLE SC

Panel 2.2

Rel. Exp. (%) Ag2615,

Rel. Exp. (%) Ag2615,

Tissue Name
Run 175128279
Tissue Name
Run 175128279

Normal Colon
6.7
Kidney Margin (OD04348)
100.0

Colon cancer (OD06064)
0.0
Kidney malignant cancer
0.0

(OD06204B)

Colon Margin (0D06064)
0.0
Kidney normal adjacent
0.0

tissue (OD06204E)

Colon cancer (OD06159)
0.0
Kidney Cancer (OD04450-
0.0

01)

Colon Margin (OD06159)
0.0
Kidney Margin (OD04450-
13.0

03)

Colon cancer (OD06297-04)
0.0
Kidney Cancer 8120613
0.0

Colon Margin (OD06297-
0.0
Kidney Margin 8120614
0.0

015)

CC Gr.2 ascend colon
0.0
Kidney Cancer 9010320
0.0

(ODO3921)

CC Margin (ODO3921)
0.0
Kidney Margin 9010321
0.0

Colon cancer metastasis
0.0
Kidney Cancer 8120607
0.0

(OD06104)

Lung Margin (OD06104)
0.0
Kidney Margin 8120608
0.0

Colon mets to lung
4.9
Normal Uterus
40.9

(OD04451-01)

Lung Margin (OD04451-02)
27.2
Uterine Cancer 064011
11.6

Normal Prostate
19.2
Normal Thyroid
0.0

Prostate Cancer (OD04410)
13.3
Thyroid Cancer 064010
0.0

Prostate Margin (OD04410)
22.7
Thyroid Cancer A302152
24.7

Normal Ovary
6.8
Thyroid Margin A302153
0.0

Ovarian cancer (OD06283-
0.0
Normal Breast
25.5

03)

Ovarian Margin (OD06283-
19.8
Breast Cancer (OD04566)
0.0

07)

Ovarian Cancer 064008
0.0
Breast Cancer 1024
0.0

Ovarian cancer (OD06145)
0.0
Breast Cancer (OD04590-
0.0

01)

Ovarian Margin (OD06145)
11.1
Breast Cancer Mets
4.7

(OD04590-03)

Ovarian cancer (OD06455-
0.0
Breast Cancer Metastasis
0.0

03)

(OD04655-05)

Ovarian Margin (OD06455-
9.0
Breast Cancer 064006
0.0

07)

Normal Lung
12.8
Breast Cancer 9100266
0.0

Invasive poor diff. lung
19.2
Breast Margin 9100265
0.0

adeno (ODO4945-01

Lung Margin (ODO4945-03)
13.2
Breast Cancer A209073
0.0

Lung Malignant Cancer
0.0
Breast Margin A2090734
20.6

(OD03126)

Lung Margin (OD03126)
0.0
Breast cancer (OD06083)
27.0

Lung Cancer (OD05014A)
0.0
Breast cancer node
25.2

metastasis (OD06083)

Lung Margin (OD05014B)
8.8
Normal Liver
15.2

Lung cancer (OD06081)
4.7
Liver Cancer 1026
0.0

Lung Margin (OD06081)
13.1
Liver Cancer 1025
6.5

Lung Cancer (OD04237-01)
0.0
Liver Cancer 6004-T
0.0

Lung Margin (OD04237-02)
0.0
Liver Tissue 6004-N
3.1

Ocular Melanoma Metastasis
0.0
Liver Cancer 6005-T
0.0

Ocular Melanoma Margin
0.0
Liver Tissue 6005-N
5.4

(Liver)

Melanoma Metastasis
0.0
Liver Cancer 064003
34.9

Melanoma Margin (Lung)
63.3
Normal Bladder
12.4

Normal Kidney
8.2
Bladder Cancer 1023
12.0

Kidney Ca, Nuclear grade 2
33.0
Bladder Cancer A302173
0.0

(OD04338)

Kidney Margin (OD04338)
4.4
Normal Stomach
14.4

Kidney Ca Nuclear grade 1/2
38.2
Gastric Cancer 9060397
0.0

(OD04339)

Kidney Margin (OD04339)
0.0
Stomach Margin 9060396
0.0

Kidney Ca, Clear cell type
0.0
Gastric Cancer 9060395
0.0

(OD04340)

Kidney Margin (OD04340)
18.0
Stomach Margin 9060394
0.0

Kidney Ca, Nuclear grade 3
0.0
Gastric Cancer 064005
7.1

(OD04348)

[0591]

88

TABLE SD

Panel 4D

Rel. Exp. (%) Ag2615,

Rel. Exp. (%) Ag2615,

Tissue Name
Run 164395568
Tissue Name
Run 164395568

Secondary Th1 act
0.0
HUVEC IL-1beta
0.0

Secondary Th2 act
0.0
HUVEC IFN gamma
2.9

Secondary Tr1 act
0.0
HUVEC TNF alpha + IFN
0.0

gamma

Secondary Th1 rest
0.8
HUVEC TNF alpha + IL4
0.0

Secondary Th2 rest
0.0
HUVEC IL-11
1.5

Secondary Tr1 rest
0.9
Lung Microvascular EC none
0.0

Primary Th1 act
0.0
Lung Microvascular EC
0.0

TNF alpha + IL-1beta

Primary Th2 act
0.0
Microvascular Dermal EC none
0.0

Primary Tr1 act
0.9
Microsvasular Dermal EC
0.0

TNF alpha + IL-1beta

Primary Th1 rest
2.6
Bronchial epithelium TNF
0.0

alpha + IL1beta

Primary Th2 rest
1.8
Small airway epithelium none
1.0

Primary Tr1 rest
1.9
Small airway epithelium
0.8

TNF alpha + IL-1beta

CD45RA CD4 lymphocyte
0.0
Coronery artery SMC rest
0.0

act

CD45RO CD4 lymphocyte
2.2
Coronery artery SMC TNF
0.0

act

alpha + IL-1beta

CD8 lymphocyte act
0.0
Astrocytes rest
0.0

Secondary CD8
2.1
Astrocytes TNF alpha + IL-
0.0

lymphocyte rest

1beta

Secondary CD8
0.0
KU-812 (Basophil) rest
0.0

lymphocyte act

CD4 lymphocyte none
0.9
KU-812 (Basophil)
1.1

PMA/ionomycin

2ry Th1/Th2/Tr1_anti-
2.2
CCD1106 (Keratinocytes) none
0.0

CD95 CH11

LAK cells rest
0.0
CCD1106 (Keratinocytes)
0.0

TNF alpha + IL-1beta

LAK cells IL-2
2.9
Liver cirrhosis
8.9

LAK cells IL-2 + IL-12
1.3
Lupus kidney
11.9

LAK cells IL-2 + IFN
0.0
NCI-H292 none
2.9

gamma

LAK cells IL-2 + IL-18
2.8
NCI-H292 IL-4
1.9

LAK cells
0.0
NCI-H292 IL-9
1.2

PMA/ionomycin

NK Cells IL-2 rest
2.4
NCI-H292 IL-13
0.0

Two Way MLR 3 day
2.1
NCI-H292 IFN gamma
1.5

Two Way MLR 5 day
0.6
HPAEC none
1.2

Two Way MLR 7 day
0.5
HPAEC TNF alpha + IL-1beta
0.0

PBMC rest
0.8
Lung fibroblast none
0.9

PBMC PWM
1.0
Lung fibroblast TNF alpha + IL-
1.0

1beta

PBMC PHA-L
0.9
Lung fibroblast IL-4
2.0

Ramos (B cell) none
34.6
Lung fibroblast IL-9
0.0

Ramos (B cell) ionomycin
100.0
Lung fibroblast IL-13
0.0

B lymphocytes PWM
0.0
Lung fibroblast IFN gamma
1.7

B lymphocytes CD40L
0.9
Dermal fibroblast CCD1070 rest
1.1

and IL-4

EOL-1 dbcAMP
0.0
Dermal fibroblast CCD1070
0.0

TNF alpha

EOL-1 dbcAMP
0.0
Dermal fibroblast CCD1070 IL-
1.2

PMA/ionomycin

1beta

Dendritic cells none
0.0
Dermal fibroblast IFN gamma
0.0

Dendritic cells LPS
1.1
Dermal fibroblast IL-4
0.0

Dendritic cells anti-CD40
0.0
IBD Colitis 2
1.2

Monocytes rest
0.0
IBD Crohn's
0.0

Monocytes LPS
0.0
Colon
1.0

Macrophages rest
0.0
Lung
3.0

Macrophages LPS
0.0
Thymus
21.5

HUVEC none
0.0
Kidney
15.1

HUVEC starved
0.0

[0592] Panel 1.3D Summary: Ag2615 Highest expression of the CG56808-01 gene is seen in a colon cancer cell line (CT=28.8). There is also significant expression in cell lines derived from lung cancer, colon cancer, and melanoma. Overall, expression appears to be more highly associated with the cancer cell lines than with the normal tissues. Thus, expression of this gene could be used to differentiate between cell lines derived from these cancers and other samples on this panel. Furthermore, expression of this gene could also be used as a marker to detect the presence of these cancers.

[0593] Panel 2.2 Summary: Ag2615 Highest expression of the CG56808-01 gene is seen in a sample derived from normal kidney adjacent to a kidney tumor (CT=32.9). Thus, expression of this gene could be used to differentiate between that sample and other samples on this panel and to differentiate between normal and cancerous kidney. Furthermore, therapeutic modulation of the expression or function of this gene could be beneficial in the treatment of kidney cancer.

[0594] Panel 4D Summary: Ag2615 The highest expression of the CG56808-01 gene is detected in Ramos B cells stimulated with ionomycin (CT=31.0). Lower but still significant levels of expression are seen in untreated Ramos B cells kidney from a lupus patient, liver cirrhosis, normal thymus and normal kidney tissue samples. B cells represent a principle component of immunity and contribute to the immune response in a number of important functional roles, including antibody production. For example, production of antibodies against self-antigens is a major component in autoimmune disorders such a systemic lupus erythematosus, with B cells playing a major role. Since B cells play an important role in autoimmunity, inflammatory processes and inflammatory cascades, therapeutic modulation of this gene product may reduce or eliminate the symptoms of patients suffering from asthma, allergies, chronic obstructive pulmonary disease, emphysema, Crohn's disease, ulcerative colitis, rheumatoid arthritis, psoriasis, osteoarthritis, and other autoimmune disorders including systemic lupus erythematosus.

[0595] T. GMAC027641_A: GPCR

[0596] Expression of gene GMAC027641_A was assessed using the primer-probe set Ag2608, described in Table TA. Results of the RTQ-PCR runs are shown in Tables TB, TC, and TD.

89TABLE TAProbe Name Ag2608StartSEQ IDPrimersSequencesLengthPositionNO:Forward5′-tgccacattcatgtatgtcttg-3′22781250ProbeTET-5′-cacagccccaaacaagacaacatcat-3′-TAMRA26815251Reverse5′-ggctggagtgacaattgtgtag-3′22850252

[0597]

90

TABLE TB

CNS_neurodegeneration_v1.0

Rel. Exp. (%) Ag2608, Run

Rel. Exp. (%) Ag2608, Run

Tissue Name
208393678
Tissue Name
208393678

AD 1 Hippo
10.2
Control (Path) 3
7.7

Temporal Ctx

AD 2 Hippo
29.9
Control (Path) 4
56.3

Temporal Ctx

AD 3 Hippo
2.0
AD 1 Occipital Ctx
26.4

AD 4 Hippo
15.8
AD 2 Occipital Ctx
0.0

(Missing)

AD 5 Hippo
73.7
AD 3 Occipital Ctx
11.7

AD 6 Hippo
16.7
AD 4 Occipital Ctx
15.7

Control 2 Hippo
18.4
AD 5 Occipital Ctx
26.6

Control 4 Hippo
1.7
AD 6 Occipital Ctx
21.9

Control (Path) 3 Hippo
2.9
Control 1 Occipital Ctx
0.0

AD 1 Temporal Ctx
10.1
Control 2 Occipital Ctx
32.5

AD 2 Temporal Ctx
39.0
Control 3 Occipital Ctx
48.6

AD 3 Temporal Ctx
2.9
Control 4 Occipital Ctx
2.8

AD 4 Temporal Ctx
30.1
Control (Path) 1
100.0

Occipital Ctx

AD 5 Inf Temporal Ctx
61.6
Control (Path) 2
45.7

Occipital Ctx

AD 5 Sup Temporal
24.8
Control (Path) 3
2.5

Ctx

Occipital Ctx

AD 6 Inf Temporal Ctx
48.6
Control (Path) 4
40.6

Occipital Ctx

AD 6 Sup Temporal
15.2
Control 1 Parietal Ctx
8.8

Ctx

Control 1 Temporal Ctx
1.5
Control 2 Parietal Ctx
43.2

Control 2 Temporal Ctx
30.8
Control 3 Parietal Ctx
40.9

Control 3 Temporal Ctx
51.4
Control (Path) 1
60.3

Parietal Ctx

Control 3 Temporal Ctx
7.5
Control (Path) 2
66.0

Parietal Ctx

Control (Path) 1
66.9
Control (Path) 3
8.1

Temporal Ctx

Parietal Ctx

Control (Path) 2
58.2
Control (Path) 4
56.3

Temporal Ctx

Parietal Ctx

[0598]

91

TABLE TC

Panel 2.1

Rel. Exp. (%) Ag2608,

Rel. Exp. (%) Ag2608,

Tissue Name
Run 170686178
Tissue Name
Run 170686178

Normal Colon
13.2
Kidney Cancer 9010320
0.0

Colon cancer (OD06064)
14.7
Kidney margin 9010321
82.9

Colon cancer margin
39.2
Kidney Cancer 8120607
16.5

(OD06064)

Colon cancer (OD06159)
0.0
Kidney margin 8120608
0.0

Colon cancer margin
13.3
Normal Uterus
68.8

(OD06159)

Colon cancer (OD06298-08)
0.0
Uterus Cancer
0.0

Colon cancer margin
12.9
Normal Thyroid
10.3

(OD06298-018)

Colon Cancer Gr.2 ascend colon
16.7
Thyroid Cancer
0.0

(ODO3921)

Colon Cancer margin
18.4
Thyroid Cancer
0.0

(ODO3921)

A302152

Colon cancer metastasis
25.2
Thyroid margin
0.0

(OD06104)

A302153

Lung margin (OD06104)
11.7
Normal Breast
30.1

Colon mets to lung (OD04451-
0.0
Breast Cancer
0.0

01)

Lung margin (OD04451-02)
11.7
Breast Cancer
26.1

Normal Prostate
0.0
Breast Cancer
0.0

(OD04590-01)

Prostate Cancer (OD04410)
10.3
Breast Cancer Mets
0.0

(OD04590-03)

Prostate margin (OD04410)
0.0
Breast Cancer
0.0

Metastasis

Normal Lung
11.4
Breast Cancer
0.0

Invasive poor diff. lung adeno 1
27.7
Breast Cancer 9100266
0.0

(ODO4945-01)

Lung margin (ODO4945-03)
0.0
Breast margin 9100265
0.0

Lung Malignant Cancer
0.0
Breast Cancer A209073
16.5

(OD03126)

Lung margin (OD03126)
0.0
Breast margin
13.1

A2090734

Lung Cancer (OD05014A)
0.0
Normal Liver
0.0

Lung margin (OD05014B)
23.7
Liver Cancer 1026
0.0

Lung Cancer (OD04237-01)
11.7
Liver Cancer 1025
0.0

Lung margin (OD04237-02)
0.0
Liver Cancer 6004-T
0.0

Ocular Mel Met to Liver
0.0
Liver Tissue 6004-N
7.1

(ODO4310)

Liver margin (ODO4310)
0.0
Liver Cancer 6005-T
0.0

Melanoma Mets to Lung
20.6
Liver Tissue 6005-N
0.0

(OD04321)

Lung margin (OD04321)
0.0
Liver Cancer
0.0

Normal Kidney
0.0
Normal Bladder
19.6

Kidney Ca, Nuclear grade 2
13.6
Bladder Cancer
0.0

(OD04338)

Kidney margin (OD04338)
15.6
Bladder Cancer
0.0

Kidney Ca Nuclear grade 1/2
24.0
Normal Ovary
0.0

(OD04339)

Kidney margin (OD04339)
0.0
Ovarian Cancer
0.0

Kidney Ca, Clear cell type
0.0
Ovarian cancer
0.0

(OD04340)

(OD06145)

Kidney margin (OD04340)
0.0
Ovarian cancer margin
100.0

(OD06145)

Kidney Ca, Nuclear grade 3
0.0
Normal Stomach
12.9

(OD04348)

Kidney margin (OD04348)
0.0
Gastric Cancer 9060397
0.0

Kidney Cancer (OD04450-01)
52.9
Stomach margin
0.0

9060396

Kidney margin (OD04450-03)
12.0
Gastric Cancer 9060395
0.0

Kidney Cancer 8120613
0.0
Stomach margin
23.3

9060394

Kidney margin 8120614
0.0
Gastric Cancer 064005
0.0

[0599]

92

TABLE TD

Panel 4D

Rel. Exp. (%) Ag2608,

Rel. Exp. (%) Ag2608,

Tissue Name
Run 164205024
Tissue Name
Run 164205024

Secondary Th1 act
22.5
HUVEC IL-1beta
7.3

Secondary Th2 act
72.2
HUVEC IFN gamma
7.1

Secondary Tr1 act
25.3
HUVEC TNF alpha + IFN
6.5

gamma

Secondary Th1 rest
6.0
HUVEC TNF alpha + IL4
8.1

Secondary Th2 rest
3.4
HUVEC IL-11
0.0

Secondary Tr1 rest
0.0
Lung Microvascular EC none
13.8

Primary Th1 act
51.4
Lung Microvascular EC
0.0

TNF alpha + IL-1beta

Primary Th2 act
29.5
Microvascular Dermal EC none
3.2

Primary Tr1 act
30.6
Microsvasular Dermal EC
0.0

TNF alpha + IL-1beta

Primary Th1 rest
8.8
Bronchial epithelium TNF
0.0

alpha + IL1beta

Primary Th2 rest
10.7
Small airway epithelium none
0.0

Primary Tr1 rest
11.8
Small airway epithelium
17.6

TNF alpha + IL-1beta

CD45RA CD4 lymphocyte
5.8
Coronery artery SMC rest
3.1

act

CD45RO CD4 lymphocyte
10.2
Coronery artery SMC TNF
3.3

act

alpha + IL-1beta

CD8 lymphocyte act
0.0
Astrocytes rest
3.1

Secondary CD8
2.0
Astrocytes TNF alpha + IL-
6.4

lymphocyte rest

1beta

Secondary CD8
6.5
KU-812 (Basophil) rest
0.0

lymphocyte act

CD4 lymphocyte none
0.0
KU-812 (Basophil)
21.0

PMA/ionomycin

2ry Th1/Th2/Tr1_anti-
0.0
CCD1106 (Keratinocytes) none
9.9

CD95 CH11

LAK cells rest
3.4
CCD1106 (Keratinocytes)
0.0

TNF alpha + IL-1beta

LAK cells IL-2
0.0
Liver cirrhosis
10.3

LAK cells IL-2 + IL-12
0.4
Lupus kidney
10.5

LAK cells IL-2 + IFN
0.0
NCI-H292 none
44.8

gamma

LAK cells IL-2 + IL-18
0.0
NCI-H292 IL-4
44.1

LAK cells
0.0
NCI-H292 IL-9
24.1

PMA/ionomycin

NK Cells IL-2 rest
0.0
NCI-H292 IL-13
8.5

Two Way MLR 3 day
0.0
NCI-H292 IFN gamma
39.2

Two Way MLR 5 day
0.0
HPAEC none
12.5

Two Way MLR 7 day
0.0
HPAEC TNF alpha + IL-1beta
6.3

PBMC rest
0.0
Lung fibroblast none
0.0

PBMC PWM
0.0
Lung fibroblast TNF alpha + IL-
3.1

1beta

PBMC PHA-L
17.6
Lung fibroblast IL-4
12.5

Ramos (B cell) none
43.8
Lung fibroblast IL-9
5.8

Ramos (B cell) ionomycin
100.0
Lung fibroblast IL-13
1.4

B lymphocytes PWM
3.6
Lung fibroblast IFN gamma
7.7

B lymphocytes CD40L
0.0
Dermal fibroblast CCD1070 rest
49.3

and IL-4

EOL-1 dbcAMP
0.0
Dermal fibroblast CCD1070
24.8

TNF alpha

EOL-1 dbcAMP
0.0
Dermal fibroblast CCD1070 IL-
13.9

PMA/ionomycin

1beta

Dendritic cells none
0.0
Dermal fibroblast IFN gamma
16.3

Dendritic cells LPS
0.0
Dermal fibroblast IL-4
0.0

Dendritic cells anti-CD40
0.0
IBD Colitis 2
0.0

Monocytes rest
0.0
IBD Crohn's
0.0

Monocytes LPS
0.0
Colon
12.7

Macrophages rest
0.0
Lung
3.1

Macrophages LPS
3.4
Thymus
13.9

HUVEC none
8.8
Kidney
34.6

HUVEC starved
10.4

[0600] CNS_neurodegeneration_v1.0 Summary: Ag2608 The GMAC027641-A gene represents a novel G-protein coupled receptor (GPCR) with expression in the brain. The GPCR family of receptors contains a large number of neurotransmitter receptors, including the dopamine, serotonin, a and b-adrenergic, acetylcholine muscarinic, histamine, peptide, and metabotropic glutamate receptors. GPCRs are excellent drug targets in various neurologic and psychiatric diseases. All antipsychotics have been shown to act at the dopamine D2 receptor; similarly novel antipsychotics also act at the serotonergic receptor, and often the muscarinic and adrenergic receptors as well. While the majority of antidepressants can be classified as selective serotonin reuptake inhibitors, blockade of the 5-HT1A and a2 adrenergic receptors increases the effects of these drugs. The GPCRs are also of use as drug targets in the treatment of stroke. Blockade of the glutamate receptors may decrease the neuronal death resulting from excitotoxicity; further more the purinergic receptors have also been implicated as drug targets in the treatment of cerebral ischemia. The b-adrenergic receptors have been implicated in the treatment of ADHD with Ritalin, while the a-adrenergic receptors have been implicated in memory. Therefore this gene may be of use as a small molecule target for the treatment of any of the described diseases.

[0601] Furthermore, this GPCR appears to be down-regulated in the temporal cortex of Alzheimer's disease patients. Therefore, up-regulation of this gene or its protein product, or treatment with specific agonists for this receptor may be of use in reversing the dementia/memory loss associated with this disease and neuronal death (El Yacoubi et al., Adenosine A2A receptor antagonists are potential antidepressants: evidence based on pharmacology and A2A receptor knockout mice. Br J Pharmacol 134(1):68-77, 2001; Blier, Pharmacology of rapid-onset antidepressant treatment strategies. Clin Psychiatry 62 Suppl 15:12-7, 2001; Tranquillini and Reggiani, Glycine-site antagonists and stroke. Expert Opin Investig Drugs 8(11):1837-1848, 1999; Monopoli et al., Blockade of adenosine A2A receptors by SCH 58261 results in neuroprotective effects in cerebral ischaemia in rats. Neuroreport 9(17):3955-9, 1998).

[0602] Panel 1.3D Summary: Ag2608 Expression of the GMAC027641_A gene is low/undetectable (CTs>35) across all of the samples on this panel (data not shown).

[0603] Panel 2.1 Summary: Ag2608 The expression of the GMAC027641-A gene is largely restricted to normal tissue samples, with highest expression in the ovary (CT=33.3). In addition, normal uterus, normal kidney, normal colon and a sample derived from malignant kidney all show substantial expression of this gene. Thus, the expression of this gene could be used to distinguish these listed samples from the other samples in the panel. Moreover, therapeutic modulation of this gene, throught the use of small molecule drugs, antibodies or protein therapeutics might be of benefit for the treatment of kidney cancer.

[0604] Panel 4D Summary: Ag2608 The GMAC027641_A transcript is expressed in polarized T cells (Th1, Th2, Tr1), activated Ramos B lymphoma, the NCI-H292 tumor line and the dermal fibrolblast cell line CCD1070. The transcript appears to induced by T cell differentiation and active proliferation. Proliferation and activation in the absence of polarizing agents, for example with CD45RA or CD45RO T cells, is not sufficient for expression. Tumor lines and cell lines such as NCI-H292 cells, Ramos B cells and CCD1070 also express this transcript regardless of treatment. The expression pattern of this transcript in T cells and its putative role as a GPCR suggests that it may therefore be important in T cell polarization. Thus, therapeutic regulation of the transcript or the protein encoded by the transcript could be important in immune modulation and in the treatment of T cell-mediated diseases such as asthma, arthritis, psoriasis, IBD, and lupus.

[0605] U. CG56290-01 and GMAC027522 B and GMAC036216 D and GMAC009775 A: Olfactory Receptor

[0606] Expression of gene and full length physical clone CG56290-01 was assessed using the primer-probe sets Ag2609, Ag2611 and Ag1500, described in Tables UA, UB and UC. Results of the RTQ-PCR runs are shown in Tables UD, UE, UF, UG, UH and UI.

93TABLE UAProbe Name Ag2609StartSEQ IDPrimersSequencesLengthPositionNO:Forward5′-cattgtgattgtctgtgtggat-3′22138253ProbeTET-5′-tcttcctcagccacctctctaccctg-3′-TAMRA26185254Reverse5′-ttatggttgtgaccaggatctc-3′22211255

[0607]

94

TABLE UB

Probe Name Ag2611

Start
SEQ ID

Primers
Sequences
Length
Position
NO:

Forward
5′-tgattgtctgtgtggataaacg-3′
22
143
256

Probe
TET-5′-tcttcctcagccacctctctaccctg-3′-TAMRA
26
185
257

Reverse
5′-ttatggttgtgaccaggatctc-3′
22
211
258

[0608]

95

TABLE UC

Probe Name Ag1500

Start
SEQ ID

Primers
Sequences
Length
Position
NO:

Forward
5′-tgattgtctgtgtggataaacg-3′
22
143
259

Probe
TET-5′-tcttcctcagccacctctctaccctg-3′-TAMRA
26
185
260

Reverse
5′-ttatggttgtgaccaggatctc-3′
22
211
261

[0609]

96

TABLE UD

CNS_neurodegeneration_v1.0

Rel. Exp. (%)
Rel. Exp. (%)

Rel. Exp. (%)
Rel. Exp. (%)

Ag2609, Run
Ag2611, Run

Ag2609, Run
Ag2611, Run

Tissue Name
208971592
208971593
Tissue Name
208971592
208971593

AD 1 Hippo
12.7
2.2
Control (Path)
0.0
0.8

3 Temporal Ctx

AD 2 Hippo
39.5
11.1
Control (Path)
12.7
2.5

4 Temporal Ctx

AD 3 Hippo
7.3
0.9
AD 1 Occipital
12.8
100.0

Ctx

AD 4 Hippo
4.2
0.8
AD 2 Occipital
0.0
0.0

Ctx (Missing)

AD 5 Hippo
36.3
14.0
AD 3 Occipital
6.2
1.8

Ctx

AD 6 Hippo
63.7
11.3
AD 4 Occipital
9.7
3.9

Ctx

Control 2
11.3
3.5
AD 5 Occipital
13.4
4.0

Hippo

Ctx

Control 4
2.5
0.9
AD 5 Occipital
19.9
4.7

Hippo

Ctx

Control (Path)
5.0
1.1
Control 1
15.9
2.0

3 Hippo

Occipital Ctx

AD 1
21.3
7.5
Control 2
24.7
7.0

Temporal Ctx

Occipital Ctx

AD 2
31.2
12.1
Control 3
8.8
4.5

Temporal Ctx

Occipital Ctx

AD 3
9.5
1.9
Control 4
4.5
2.9

Temporal Ctx

Occipital Ctx

AD 4
28.1
3.7
Control (Path)
100.0
17.8

Temporal Ctx

1 Occipital Ctx

AD 5 Inf
74.7
13.6
Control (Path)
17.9
5.8

Temporal Ctx

2 Occipital Ctx

AD 5 Sup
18.2
4.2
Control (Path)
2.8
1.8

Temporal Ctx

3 Occipital Ctx

AD 6 Inf
54.7
16.2
Control (Path)
12.3
6.7

Temporal Ctx

4 Occipital Ctx

AD 6 Sup
39.8
13.6
Control 1
20.4
8.1

Temporal Ctx

Parietal Ctx

Control 1
3.2
1.0
Control 2
19.3
4.8

Temporal Ctx

Parietal Ctx

Control 2
7.1
3.1
Control 3
30.4
6.8

Temporal Ctx

Parietal Ctx

Control 3
8.2
2.6
Control (Path)
40.9
11.5

Temporal Ctx

1 Parietal Ctx

Control 3
6.3
5.1
Control (Path)
21.2
6.3

Temporal Ctx

2 Parietal Ctx

Control (Path)
34.6
9.7
Control (Path)
3.7
0.7

1 Temporal

3 Parietal Ctx

Ctx

Control (Path)
19.6
5.1
Control (Path)
32.1
8.5

2 Temporal

4 Parietal Ctx

Ctx

[0610]

97

TABLE UE

Panel 1.2

Rel. Exp. (%) Ag1500, Run

Rel. Exp. (%) Ag1500, Run

Tissue Name
141889923
Tissue Name
141889923

Endothelial cells
3.1
Renal ca. 786-0
8.0

Heart (Fetal)
2.7
Renal ca. A498
11.5

Pancreas
0.0
Renal ca. RXF 393
3.6

Pancreatic ca. CAPAN 2
2.8
Renal ca. ACHN
4.2

Adrenal Gland
8.4
Renal ca. UO-31
8.0

Thyroid
0.0
Renal ca. TK-10
24.5

Salivary gland
39.8
Liver
0.0

Pituitary gland
1.4
Liver (fetal)
1.1

Brain (fetal)
1.2
Liver ca. (hepatoblast)
1.9

HepG2

Brain (whole)
5.0
Lung
0.0

Brain (amygdala)
4.3
Lung (fetal)
1.0

Brain (cerebellum)
13.7
Lung ca. (small cell) LX-1
30.4

Brain (hippocampus)
31.4
Lung ca. (small cell)
65.5

NCI-H69

Brain (thalamus)
50.7
Lung ca. (s. cell var.)
1.5

SHP-77

Cerebral Cortex
100.0
Lung ca. (large cell)NCI-
23.2

H460

Spinal cord
18.3
Lung ca. (non-sm. cell)
13.9

A549

glio/astro U87-MG
12.1
Lung ca. (non-s. cell)
31.4

NCI-H23

glio/astro U-118-MG
0.0
Lung ca. (non-s. cell)
28.5

HOP-62

astrocytoma SW1783
11.6
Lung ca. (non-s. cl) NCI-
51.1

H522

neuro*; met SK-N-AS
8.5
Lung ca. (squam.) SW
4.3

900

astrocytoma SF-539
1.1
Lung ca. (squam.) NCI-
22.4

H596

astrocytoma SNB-75
22.4
Mammary gland
18.2

glioma SNB-19
19.1
Breast ca.* (pl. ef) MCF-7
4.5

glioma U251
2.4
Breast ca.* (pl. ef) MDA-
0.0

MB-231

glioma SF-295
12.2
Breast ca.* (pl. ef) T47D
91.4

Heart
15.5
Breast ca. BT-549
10.6

Skeletal Muscle
7.0
Breast ca. MDA-N
63.3

Bone marrow
1.7
Ovary
7.9

Thymus
0.9
Ovarian ca. OVCAR-3
8.1

Spleen
0.0
Ovarian ca. OVCAR-4
20.2

Lymph node
0.0
Ovarian ca. OVCAR-5
95.9

Colorectal
6.2
Ovarian ca. OVCAR-8
88.3

Stomach
0.0
Ovarian ca. IGROV-1
24.7

Small intestine
2.4
Ovarian ca. (ascites) SK-
16.4

OV-3

Colon ca. SW480
1.1
Uterus
1.1

Colon ca.* SW620
5.8
Placenta
27.7

(SW480 met)

Colon ca. HT29
14.2
Prostate
3.2

Colon ca. HCT-116
3.2
Prostate ca.* (bone met)
14.5

PC-3

Colon ca. CaCo-2
10.8
Testis
0.0

CC Well to Mod Diff
17.8
Melanoma Hs688(A).T
2.3

(ODO3866)

Colon ca. HCC-2998
6.9
Melanoma* (met)
12.3

Hs688(B).T

Gastric ca. (liver met)
12.2
Melanoma UACC-62
54.7

NCI-N87

Bladder
17.4
Melanoma M14
89.5

Trachea
0.0
Melanoma LOX IMVI
0.0

Kidney
14.9
Melanoma* (met) SK-
21.5

MEL-5

Kidney (fetal)
16.7

[0611]

98

TABLE UF

Panel 1.3D

Rel. Exp. (%)
Rel. Exp. (%)

Rel. Exp. (%)
Rel. Exp. (%)

Ag2609, Run
Ag2611, Run

Ag2609, Run
Ag2611, Run

Tissue Name
166219826
166190369
Tissue Name
166219826
166190369

Liver
0.0
0.0
Kidney (fetal)
0.0
0.0

adenocarcinoma

Pancreas
0.0
0.0
Renal ca. 786-0
0.0
0.0

Pancreatic ca.
0.0
7.0
Renal ca. A498
0.0
0.0

CAPAN 2

Adrenal gland
12.9
0.0
Renal ca. RXF
12.7
16.5

393

Thyroid
0.0
0.0
Renal ca. ACHN
0.0
0.0

Salivary gland
6.2
0.0
Renal ca. UO-31
0.0
0.0

Pituitary gland
0.0
0.0
Renal ca. TK-10
7.4
0.0

Brain (fetal)
26.4
0.0
Liver
0.0
0.0

Brain (whole)
26.2
26.8
Liver (fetal)
0.0
0.0

Brain (amygdala)
3.9
5.4
Liver ca.
0.0
8.0

(hepatoblast)

HepG2

Brain (cerebellum)
0.0
6.6
Lung
0.0
0.0

Brain
17.2
6.2
Lung (fetal)
0.0
0.0

(hippocampus)

Brain (substantia
31.9
78.5
Lung ca. (small
19.3
2.6

nigra)

cell) LX-1

Brain (thalamus)
82.4
100.0
Lung ca. (small
0.0
0.0

cell) NCI-H69

Cerebral Cortex
0.0
19.2
Lung ca. (s. cell
0.0
0.0

var.) SHP-77

Spinal cord
100.0
100.0
Lung ca. (large
0.0
0.0

cell)NCI-H460

glio/astro U87-MG
0.0
0.0
Lung ca. (non-
0.0
0.0

sm. cell) A549

glio/astro U-118-
0.0
0.0
Lung ca. (non-
10.2
0.0

MG

s. cell) NCI-H23

astrocytoma
6.9
0.0
Lung ca. (non-
0.0
6.2

SW1783

s. cell) HOP-62

neuro*; met SK-N-
0.0
0.0
Lung ca. (non-
0.0
0.0

AS

s. cl) NCI-H522

astrocytoma SF-539
0.0
0.0
Lung ca.
0.0
0.0

(squam.) SW 900

astrocytoma SNB-
0.0
0.0
Lung ca.
0.0
0.0

75

(squam.) NCI-

H596

glioma SNB-19
5.7
5.7
Mammary gland
0.0
0.0

glioma U251
0.0
0.0
Breast ca.*
0.0
0.0

(pl. ef) MCF-7

glioma SF-295
6.7
0.0
Breast ca.*
0.0
0.0

(pl. ef) MDA-

MB-231

Heart (Fetal)
0.0
6.1
Breast ca.* (pl.
22.1
32.5

ef) T47D

Heart
0.0
0.0
Breast ca. BT-
0.0
0.0

549

Skeletal muscle
0.0
0.0
Breast ca. MDA-N
3.2
29.7

(Fetal)

Skeletal muscle
0.0
0.0
Ovary
0.0
0.0

Bone marrow
0.0
0.0
Ovarian ca.
0.0
0.0

OVCAR-3

Thymus
0.0
0.0
Ovarian ca.
0.0
0.0

OVCAR-4

Spleen
0.0
0.0
Ovarian ca.
10.7
0.0

OVCAR-5

Lymph node
0.0
0.0
Ovarian ca.
6.0
32.8

OVCAR-8

Colorectal
6.7
0.0
Ovarian ca.
0.0
0.0

IGROV-1

Stomach
0.0
0.0
Ovarian ca.
8.8
0.0

(ascites) SK-OV-3

Small intestine
0.0
15.4
Uterus
0.0
0.0

Colon ca. SW480
0.0
0.0
Placenta
38.7
27.9

Colon ca.* SW620
0.0
0.0
Prostate
0.0
0.0

(SW480 met)

Colon ca. HT29
0.0
0.0
Prostate ca.*
0.0
6.5

(bone met) PC-3

Colon ca. HCT-116
0.0
0.0
Testis
34.9
14.9

Colon ca. CaCo-2
0.0
0.0
Melanoma
0.0
0.0

Hs688(A).T

CC Well to Mod
0.0
0.0
Melanoma*
0.0
0.0

Diff (ODO3866)

(met)

Hs688(B).T

Colon ca. HCC-
0.0
0.0
Melanoma
0.0
32.8

2998

UACC-62

Gastric ca. (liver
0.0
0.0
Melanoma M14
0.0
18.4

met) NCI-N87

Bladder
0.0
0.0
Melanoma LOX
0.0
0.0

IMVI

Trachea
0.0
0.0
Melanoma*
7.9
0.0

(met) SK-MEL-5

Kidney
0.0
0.0
Adipose
0.0
0.0

[0612]

99

TABLE UG

Panel 2.2

Rel. Exp. (%)
Rel. Exp. (%)

Rel. Exp. (%)
Rel. Exp. (%)

Ag2609, Run
Ag2611, Run

Ag2609, Run
Ag2611, Run

Tissue Name
175128270
175128271
Tissue Name
175128270
175128271

Normal Colon
6.3
0.0
Kidney Margin
14.6
12.7

(OD04348)

Colon cancer
0.0
0.0
Kidney malignant
9.7
9.7

(OD06064)

cancer

(OD06204B)

Colon Margin
0.0
0.0
Kidney normal
0.0
0.0

(OD06064)

adjacent tissue
0.0
0.0

(OD06204E)

Colon cancer
0.0
0.0
Kidney Cancer
18.0
0.0

(OD06159)

(OD04450-01)

Colon Margin
5.4
0.0
Kidney Margin
0.0
0.0

(OD06159)

(OD04450-03)

Colon cancer
0.0
0.0
Kidney Cancer
0.0
0.0

(OD06297-04)

8120613

Colon Margin
6.0
0.0
Kidney Margin
0.0
0.0

(OD06297-015)

8120614

CC Gr.2 ascend
0.0
0.0
Kidney Cancer
0.0
0.0

colon (ODO3921

9010320

CC Margin
0.0
0.0
Kidney Margin
0.0
0.0

(ODO3921)

9010321

Colon cancer
0.0
0.0
Kidney Cancer
0.0
0.0

metastasis

8120607

(OD06104)

Lung Margin
0.0
0.0
Kidney Margin
0.0
0.0

(OD06104)

8120608

Colon mets to lung
0.0
0.0
Normal Uterus
9.8
14.5

(OD04451-01)

Lung Margin
0.0
0.0
Uterine Cancer
0.0
0.0

(OD04451-02)

064011

Normal Prostate
0.0
0.0
Normal Thyroid
0.0
0.0

Prostate Cancer
0.0
0.0
Thyroid Cancer
0.0
0.0

(OD04410)

Prostate Margin
0.0
0.0
Thyroid Cancer
0.0
15.1

(OD04410)

A302152

Normal Ovary
0.0
0.0
Thyroid Margin
0.0
0.0

A302153

Ovarian cancer
0.0
0.0
Normal Breast
18.7
33.9

(OD06283-03)

Ovarian Margin
7.1
11.0
Breast Cancer
0.0
0.0

(OD06283-07)

Ovarian Cancer
28.7
0.0
Breast Cancer
21.2
17.7

Ovarian cancer
0.0
5.6
Breast Cancer
0.0
0.0

(OD06145)

(OD04590-01)

Ovarian Margin
15.9
0.0
Breast Cancer Mets
8.4
0.0

(OD06145)

(OD04590-03)

Ovarian cancer
6.8
0.0
Breast Cancer
0.0
0.0

(OD06455-03)

Metastasis

Ovarian Margin
0.0
0.0
Breast Cancer
1.5
9.9

(OD06455-07)

Normal Lung
0.0
0.0
Breast Cancer
100.0
100.0

9100266

Invasive poor diff.
0.0
0.0
Breast Margin
5.9
7.8

lung adeno

9100265

(ODO4945-01

Lung Margin
3.1
0.0
Breast Cancer
3.7
7.0

(ODO4945-03)

A209073

Lung Malignant
0.0
0.0
Breast Margin
0.0
27.5

Cancer (OD03126)

A2090734

Lung Margin
0.0
0.0
Breast cancer
9.9
14.3

(OD03126)

(OD06083)

Lung Cancer
0.0
0.0
Breast cancer node
13.5
5.3

(OD05014A)

metastasis

(OD06083)

Lung Margin
0.0
4.3
Normal Liver
0.0
0.0

(OD05014B)

Lung cancer
0.0
0.0
Liver Cancer 1026
0.0
0.0

(OD06081)

Lung Margin
0.0
8.0
Liver Cancer 1025
0.0
0.0

(OD06081)

Lung Cancer
0.0
0.0
Liver Cancer 6004-T
0.0
0.0

(OD04237-01)

Lung Margin
0.0
0.0
Liver Tissue 6004-N
0.0
0.0

(OD04237-02)

Ocular Mel Met to
0.0
10.3
Liver Cancer 6005-T
5.2
0.0

Liver (ODO4310)

Liver Margin
0.0
0.0
Liver Tissue 6005-N
0.0
0.0

(ODO4310)

Melanoma
16.5
15.2
Liver Cancer
0.0
0.0

Metastasis

Lung Margin
0.0
0.0
Normal Bladder
0.0
0.0

(OD04321)

Normal Kidney
7.4
11.0
Bladder Cancer
0.0
9.0

Kidney Ca, Nuclear
4.3
0.0
Bladder Cancer
10.2
0.0

grade 2 (OD04338)

Kidney Margin
0.0
2.8
Normal Stomach
12.3
0.0

(OD04338)

Kidney Ca Nuclear
5.6
10.1
Gastric Cancer
0.0
0.0

grade 1/2

9060397

(OD04339)

Kidney Margin
0.0
7.0
Stomach Margin
0.0
0.0

(OD04339)

9060396

Kidney Ca, Clear
0.0
7.3
Gastric Cancer
11.4
0.0

cell type

9060395

(OD04340)

Kidney Margin
0.0
0.0
Stomach Margin
0.0
0.0

(OD04340)

9060394

Kidney Ca, Nuclear
0.0
0.0
Gastric Cancer
0.0
0.0

grade 3 (OD04348)

064005

[0613]

100

TABLE UH

Panel 4D

Rel.
Rel.
Rel.

Rel.
Rel.
Rel.

Exp. (%)
Exp. (%)
Exp. (%)

Exp. (%)
Exp. (%)
Exp. (%)

Ag2609,
Ag2609,
Ag2611,

Ag2609,
Ag2609,
Ag2611,

Run
Run
Run

Run
Run
Run

Tissue Name
164289991
164347907
164398661
Tissue Name
164289991
164347907
164398661

Secondary Th1 act
6.4
6.4
4.8
HUVEC IL-
0.0
0.0
0.0

1beta

Secondary Th2 act
12.7
12.7
13.9
HUVEC INF
4.9
4.9
0.0

gamma

Secondary Tr1 act
0.9
0.9
14.7
HUVEC TNF
15.3
15.3
6.9

alpha + IFN

gamma

Secondary Th1
3.9
3.9
0.0
HUVEC TNF
18.4
18.4
0.0

rest

alpha + IL4

Secondary Th2
0.0
0.0
0.0
HUVEC IL-11
0.8
0.8
6.0

rest

Secondary Tr1 rest
0.4
0.4
0.0
Lung
6.5
6.5
66.0

Microvascular

EC none

Primary Th1 act
0.0
0.0
19.6
Lung
9.5
9.5
18.3

Microvascular

EC TNF alpha +

IL-1beta

Primary Th2 act
9.2
9.2
0.0
Microvascular
0.4
0.4
0.0

Dermal EC none

Primary Tr1 act
4.2
4.2
7.1
Microvascular
16.2
16.2
3.0

Dermal EC

TNF alpha + IL-

1beta

Primary Th1 rest
16.8
16.8
20.4
Bronchial
1.3
1.3
6.3

epithelium

TNF alpha +

IL1beta

Primary Th2 rest
13.6
13.6
15.0
Small airway
2.9
2.9
0.0

epithelium none

Primary Tr1 rest
1.5
1.5
0.0
Small airway
10.9
10.9
3.9

epithelium

TNF alpha + IL-

1beta

CD45RA CD4
0.0
0.0
13.5
Coronery artery
0.0
0.0
0.0

lymphocyte act

SMC rest

CD45RO CD4
0.2
0.2
1.3
Coronery artery
0.4
0.4
5.9

lymphocyte act

SMC TNF alpha +

IL-1beta

CD8 lymphocyte
0.0
0.0
4.5
Astrocytes rest
9.9
9.9
0.0

act

Secondary CD8
13.5
13.5
4.5
Astrocytes
0.0
0.0
0.0

lymphocyte rest

TNF alpha + IL-

1beta

Secondary CD8
13.5
13.5
7.3
KU-812
3.1
3.1
4.0

lymphocyte act

(Basophil) rest

CD4 lymphocyte
10.2
10.2
10.5
KU-812
3.9
3.9
4.8

none

(Basophil)

PMA/ionomycin

2ry
16.3
16.3
5.7
CCD1106
9.6
9.6
0.0

Th1/Th2/Tr1 anti-

(Keratinocytes)

CD95 CH11

none

LAK cells rest
17.1
17.1
6.3
CCD1106
6.5
6.5
0.0

(Keratinocytes)

TNF alpha + IL-

1beta

LAK cells IL-2
0.0
0.0
0.0
Liver cirrhosis
22.5
22.5
25.9

LAK cells IL-
3.0
3.0
19.5
Lupus kidney
3.0
3.0
0.0

2 + IL-12

LAK cells IL-
7.7
7.7
0.0
NCI-H292 none
4.4
4.4
0.0

2 + IFN gamma

LAK cells IL-2 +
4.1
4.1
0.0
NCI-H292 IL-4
3.5
3.5
0.0

IL-18

LAK cells
2.0
2.0
14.2
NCI-H292 IL-9
1.5
1.5
0.0

PMA/ionomycin

NK Cells IL-2 rest
18.3
18.3
0.0
NCI-H292 IL-13
9.5
9.5
0.0

Two Way MLR 3
15.6
15.6
15.0
NCI-H292 IFN
0.0
0.0
0.0

day

gamma

Two Way MLR 5
15.8
15.8
0.0
HPAEC none
0.0
0.0
0.0

day

Two Way MLR 7
0.0
0.0
8.1
HPAEC TNF
3.2
3.2
0.0

day

alpha + IL-1beta

PBMC rest
1.6
1.6
0.0
Lung fibroblast
4.1
4.1
0.0

none

PBMC PWM
10.2
10.2
14.2
Lung fibroblast
0.0
0.0
0.0

TNF alpha + IL-

1beta

PBMC PHA-L
6.7
6.7
20.9
Lung fibroblast
5.7
5.7
0.0

IL-4

Ramos (B cell)
17.1
17.1
7.8
Lung fibroblast
0.0
0.0
0.0

none

IL-9

Ramos (B cell)
8.2
8.2
0.0
Lung fibroblast
0.0
0.0
0.0

ionomycin

IL-13

B lymphocytes
3.7
3.7
4.5
Lung fibroblast
0.0
0.0
0.0

PWM

IFN gamma

B lymphocytes
7.2
7.2
13.2
Dermal
5.0
5.0
13.6

CD40L and IL-4

fibroblast

CCD1070 rest

EOL-1 dbcAMP
9.4
9.4
2.1
Dermal
14.7
14.7
36.1

fibroblast

CCD1070 TNF

alpha

EOL-1 dbcAMP
25.0
25.0
0.0
Dermal
0.0
0.0
6.3

PMA/ionomycin

fibroblast

CCD1070 IL-

1beta

Dendritic cells
43.2
43.2
27.2
Dermal
0.0
0.0
0.0

none

fibroblast IFN

gamma

Dendritic cells
17.6
17.6
11.3
Dermal
0.0
0.0
6.3

LPS

fibroblast IL-4

Dendritic cells
24.1
24.1
5.6
IBD Colitis 2
9.3
9.3
0.0

anti-CD40

Monocytes rest
2.2
2.2
0.0
IBD Crohn's
0.0
0.0
0.0

Monocytes LPS
69.3
69.3
100.0
Colon
1.0
1.0
6.8

Macrophages rest
100.0
100.0
97.3
Lung
11.7
11.7
19.8

Macrophages LPS
14.9
14.9
8.1
Thymus
19.3
19.3
20.2

HUVEC none
0.0
0.0
0.0
Kidney
4.4
4.4
20.2

HUVEC starved
4.7
4.7
19.1

[0614]

101

TABLE UI

Panel CNS_1

Rel. Exp. (%) Ag2609, Run

Rel. Exp. (%) Ag2609, Run

Tissue Name
171664238
Tissue Name
171664238

BA4 Control
0.0
BA17 PSP
7.2

BA4 Control2
6.5
BA17 PSP2
0.0

BA4 Alzheimer's2
6.6
Sub Nigra Control
49.3

BA4 Parkinson's
7.2
Sub Nigra Control2
37.1

BA4 Parkinson's2
0.0
Sub Nigra Alzheimer's2
6.6

BA4 Huntington's
14.7
Sub Nigra Parkinson's2
19.1

BA4
0.0
Sub Nigra Huntington's
100.0

Huntington's2

BA4 PSP
6.0
Sub Nigra
7.0

Huntington's2

BA4 PSP2
8.5
Sub Nigra PSP2
4.5

BA4 Depression
12.2
Sub Nigra Depression
0.0

BA4 Depression2
5.1
Sub Nigra Depression2
4.8

BA7 Control
18.6
Glob Palladus Control
9.5

BA7 Control2
0.0
Glob Palladus Control2
6.4

BA7 Alzheimer's2
5.6
Glob Palladus
4.8

Alzheimer's

BA7 Parkinson's
3.7
Glob Palladus
13.6

Alzheimer's2

BA7 Parkinson's2
0.0
Glob Palladus
38.2

Parkinson's

BA7 Huntington's
8.8
Glob Palladus
11.9

Parkinson's2

BA7
9.5
Glob Palladus PSP
0.0

Huntington's2

BA7 PSP
15.0
Glob Palladus PSP2
0.0

BA7 PSP2
0.0
Glob Palladus
23.5

Depression

BA7 Depression
9.1
Temp Pole Control
0.0

BA9 Control
5.9
Temp Pole Control2
0.0

BA9 Control2
10.6
Temp Pole Alzheimer's
0.0

BA9 Alzheimer's
0.0
Temp Pole Alzheimer's2
0.0

BA9 Alzheimer's2
0.0
Temp Pole Parkinson's
0.0

BA9 Parkinson's
18.8
Temp Pole Parkinson's2
3.2

BA9 Parkinson's2
5.4
Temp Pole Huntington's
0.0

BA9 Huntington's
11.6
Temp Pole PSP
0.0

BA9
0.0
TemP Pole PSP2
0.0

Huntington's2

BA9 PSP
3.3
Temp Pole Depression2
0.0

BA9 PSP2
0.0
Cing Gyr Control
17.9

BA9 Depression
4.8
Cing Gyr Control2
9.3

BA9 Depression2
0.0
Cing Gyr Alzheimer's
11.9

BA17 Control
9.6
Cing Gyr Alzheimer's2
0.0

BA17 Control2
8.4
Cing Gyr Parkinson's
38.4

BA17
0.0
Cing Gyr Parkinson's2
11.8

Alzheimer's2

BA17 Parkinson's
18.4
Cing Gyr Huntington's
29.1

BA17
20.4
Cing Gyr Huntington's2
25.5

Parkinson's2

BA17
17.9
Cing Gyr PSP
33.7

Huntington's

BA17
0.0
Cing Gyr PSP2
13.7

Huntington's2

BA17 Depression
12.2
Cing Gyr Depression
17.8

BA17 Depression2
8.5
Cing Gyr Depression2
19.8

[0615] CNS_neurodegeneration_v1.0 Summary: Ag2611/Ag2609 The CG56290-01 gene is expressed more highly in the temporal cortex of Alzheimer's diseased brain than in control brain without amyloid plaques, which are diagnostic and potentially causative of Alzheimer's disease. The CG56290-01 gene encodes a protein with homology to GPCRs. GPCRs are readily targetable with drugs, and regulate many specific brain processes, including signaling processes, that are currently the target of FDA-approved pharmaceuticals that treat Alzheimer's disease, such as the cholinergic system. The major mechanisms proposed for AbetaP-induced cytotoxicity involve the loss of Ca2+ homeostasis and the generation of reactive oxygen species (ROS). The changes in Ca2+ homeostasis could be the result of changes in G-protein-driven releases of second messengers. Thus, targeting this class of molecule can have therapeutic potential in Alzheimer's disease treatment. In particular, the increased CG56290-01 gene expression in brains affected by Alzheimer's indicates potential therapeutic value to drugs that target this GPCR (Kourie, Mechanisms of amyloid beta protein-induced modification in ion transport systems: implications for neurodegenerative diseases. Cell Mol Neurobiol 21(3):173-213, 2001).

[0616] Panel 1.2 Summary: Ag1500 Highest expression of the CG56290-01 gene is seen in the cerebral cortex (CT=30.4). Among tissues active in the central nervous system, the CG56290-01 gene is also moderately expressed in the cerebellum, hippocampus, thalamus and spinal cord. Please see CNS_neurodegeneration_panel_1.0 summary for description of the potential utility of this gene in the treatment of CNS diseases.

[0617] Among tissues with metabolic function, the CG56290-01 gene is expressed at low but significant levels in samples derived from the adrenal gland, heart and skeletal muscle. Therefore, the protein encoded by the CG56290-01 gene may be important in the pathogenesis and/or treatment of disease in any or all of the above-named tissues.

[0618] The CG56290-01 gene also shows an association with cancerous cell lines and is expressed in clusters of samples derived from breast, ovarian, melanoma and lung cancer cell lines. Thus, the expression of this gene could be used to distinguish samples derived from cell lines when compared to tissues. In addition, therapeutic modulation of the CG56290-01 gene or its protein product, through the use of small molecule drugs or antibodies, might be beneficial in the treatment of ovarian cancer, breast cancer, lung cancer or melanoma.

[0619] Panel 1.3D Summary: Ag2611/Ag2609 Two experiments with two different probe/primer sets both show preferential expression of the CG56290-01 gene in tissues originating in the central nervous system, with expression seen in the spinal cord (CT=33.1) and thalamus (CT=34.1). Please see CNS_neurodegeneration_panel_v1.0 summary for description of the potential utility of this gene in the treatment of CNS diseases.

[0620] Panel 2.2 Summary: Ag2611/Ag2609 In two experiments using two different probe and primer sets, expression of the CG56290-01 gene is limited to a sample derived from a breast cancer (CT=33.2) and appears to be overexpressed in breast cancer as compared to normal adjacent tissue. This suggests that the CG56290-01 gene could be used to distinguish breast cancer samples from other samples and for the detection of breast cancer. Moreover, therapeutic inhibition of this gene, through the use of small molecule drugs or antibodies might be of use in the treatment of breast cancer.

[0621] Panel 4D Summary: Ag2611/Ag2609 The CG56290-01 gene is expressed at moderate levels in LPS-activated monocytes but not in resting monocytes. Conversely, the CG56290-01 gene is expressed at moderate levels in resting macrophages, but at low levels in activated macrophages. This pattern is evident in experiments using two different probe and primer sets that match the CG56290-01 sequence. Since circulating monocytes and tissue macrophages are both developmentally related cell types, the CG56290-01 gene could serve as a useful target for the development of small molecule drugs as well as therapeutic antibodies. Therapeutic antibodies and small molecule inhibitors that block the function of the protein encoded by the CG56290-01 gene may be useful in reducing inflammation and autoimmune disease symptoms in patients with Crohn's disease, inflammatory bowel disease, asthma, psoriasis, and rheumatoid arthritis.

[0622] Panel CNS—1 Summary: Ag2609 Expression of the CG56290-01 gene is highest in the substantia nigra of a Huntington's disease patient, indicating that this gene may participate in the genetic dysregulation associated with the neurodegeneration that occurs in this brain region. The substantia nigra is also critical to the progression of Parkinson's disease neurodegeneration. Thus, pharmacological targeting of the GPCR encoded by the CG56290-01 gene may help counter this genetic dysregulation and contribute to the restoration of normal function in Huntington's disease as well as potentially Parkinson's disease patients. Pharmacological modulation of GPCR signaling systems is the mechanism by which powerful depression therapies, such as SSRIs, exert their effect (Perrine et al., Cognitive functioning after pallidotomy for refractory Parkinson's disease. J Neurol Neurosurg Psychiatry 65(2):150-4, 1998).

[0623] V. CG149828-01/GMAP002418_A: Olfactory Receptor

[0624] Expression of gene CG 149828-01 was assessed using the primer-probe set Ag1834, described in Table VA. Results of the RTQ-PCR runs are shown in Tables VB and VC.

102TABLE VAProbe Name Ag1834StartSEQ IDPrimersSequencesLengthPositionNO:Forward5′-ggtaggaaatagcaccctcatc-3′22116262ProbeTET-5′-cctccacacacccatgtattttgtcg-3′-TAMRA26161263Reverse5′-cagagatccagaaacgacagat-3′22193264

[0625]

103

TABLE VB

General_screening_panel_v1.5

Rel. Exp. (%) Ag1834, Run

Rel. Exp. (%) Ag1834, Run

Tissue Name
228633528
Tissue Name
228633528

Adipose
0.0
Renal ca. TK-10
0.0

Melanoma* Hs688(A).T
0.0
Bladder
0.4

Melanoma* Hs688(B).T
0.0
Gastric ca. (liver met.)
0.0

NCI-N87

Melanoma* M14
6.5
Gastric ca. KATO III
0.0

Melanoma* LOXIMVI
0.0
Colon ca. SW-948
0.0

Melanoma* SK-MEL-5
0.0
Colon ca. SW480
0.0

Squamous cell
1.4
Colon ca.* (SW480 met)
0.0

carcinoma SCC-4

SW620

Testis Pool
0.0
Colon ca. HT29
0.0

Prostate ca.* (bone met)
0.0
Colon ca. HCT-116
0.0

PC-3

Prostate Pool
0.0
Colon ca. CaCo-2
0.0

Placenta
0.0
Colon cancer tissue
0.0

Uterus Pool
0.0
Colon ca. SW1116
0.0

Ovarian ca. OVCAR-3
0.0
Colon ca. Colo-205
0.0

Ovarian ca. SK-OV-3
1.8
Colon ca. SW-48
0.0

Ovarian ca. OVCAR-4
0.0
Colon Pool
0.0

Ovarian ca. OVCAR-5
1.1
Small Intestine Pool
0.0

Ovarian ca. IGROV-1
0.0
Stomach Pool
0.0

Ovarian ca. OVCAR-8
0.0
Bone Marrow Pool
0.0

Ovary
0.0
Fetal Heart
0.0

Breast ca. MCF-7
0.0
Heart Pool
2.2

Breast ca. MDA-MB-
0.0
Lymph Node Pool
0.0

231

Breast ca. BT 549
0.0
Fetal Skeletal Muscle
52.1

Breast ca. T47D
0.0
Skeletal Muscle Pool
0.0

Breast ca. MDA-N
12.1
Spleen Pool
0.0

Breast Pool
0.0
Thymus pool
10.1

Trachea
0.0
CNS cancer (glio/astro)
0.0

U87-MG

Lung
0.0
CNS cancer (glio/astro) U-
1.2

118-MG

Fetal Lung
0.0
CNS cancer (neuro;met)
0.0

SK-N-AS

Lung ca. NCI-N417
0.0
CNS cancer (astro) SF-539
0.0

Lung ca. LX-1
0.0
CNS cancer (astro) SNB-75
0.0

Lung ca. NCI-H146
0.0
CNS cancer (glio) SNB-19
0.0

Lung ca. SHP-77
0.0
CNS cancer (glio) SF-295
0.0

Lung ca. A549
0.0
Brain (Amygdala) Pool
0.0

Lung ca. NCI-H526
0.0
Brain (cerebellum)
0.0

Lung ca. NCI-H23
0.0
Brain (fetal)
2.2

Lung ca. NCI-H460
100.0
Brain (Hippocampus) Pool
0.0

Lung ca. HOP-62
0.0
Cerebral Cortex Pool
0.0

Lung ca. NCI-H522
0.0
Brain (Substantia nigra)
0.0

Pool

Liver
0.0
Brain (Thalamus) Pool
0.0

Fetal Liver
0.0
Brain (whole)
0.0

Liver ca. HepG2
0.0
Spinal Cord Pool
0.0

Kidney Pool
1.9
Adrenal Gland
0.0

Fetal Kidney
0.0
Pituitary gland Pool
0.0

Renal ca. 786-0
0.0
Salivary Gland
0.0

Renal ca. A498
0.0
Thyroid (female)
0.0

Renal ca. ACHN
0.0
Pancreatic ca. CAPAN2
0.0

Renal ca. UO-31
0.0
Pancreas Pool
0.0

[0626]

104

TABLE VC

Panel 4D

Rel. Exp. (%) Ag1834,

Rel. Exp. (%) Ag1834,

Tissue Name
Run 165810455
Tissue Name
Run 165810455

Secondary Th1 act
0.0
HUVEC IL-1 beta
0.0

Secondary Th2 act
0.0
HUVEC IFN gamma
0.0

Secondary Tr1 act
1.2
HUVEC TNF alpha + IFN
0.0

gamma

Secondary Th1 rest
1.4
HUVEC TNF alpha + IL4
0.0

Secondary Th2 rest
0.0
HUVEC IL-11
0.0

Secondary Tr1 rest
0.0
Lung Microvascular EC none
0.0

Primary Th1 act
0.0
Lung Microvascular EC
0.0

TNF alpha + IL-1 beta

Primary Th2 act
0.0
Microvascular Dermal EC none
0.0

Primary Tr1 act
0.0
Microsvasular Dermal EC
0.0

TNF alpha + IL-1 beta

Primary Th1 rest
0.0
Bronchial epithelium TNF alpha +
0.0

IL1 beta

Primary Th2 rest
0.0
Small airway epithelium none
0.0

Primary Tr1 rest
0.0
Small airway epithelium
0.0

TNF alpha + IL-1 beta

CD45RA CD4 lymphocyte
0.0
Coronery artery SMC rest
0.0

act

CD45RO CD4 lymphocyte
0.0
Coronery artery SMC TNF alpha +
0.0

act

IL-1 beta

CD8 lymphocyte act
0.0
Astrocytes rest
0.0

Secondary CD8
0.0
Astrocytes TNF alpha + IL-1 beta
0.0

lymphocyte rest

Secondary CD8
0.0
KU-812 (Basophil) rest
0.0

lymphocyte act

CD4 lymphocyte none
0.0
KU-812 (Basophil)
0.0

PMA/ionomycin

2ry Th1/Th2/Tr1_anti-
0.0
CD1106 (Keratinocytes) none
2.0

CD95 CH11

LAK cells rest
0.0
CCD1106 (Keratinocytes)
1.0

TNF alpha + IL-1 beta

LAK cells IL-2
0.0
Liver cirrhosis
100.0

LAK cells IL-2 + IL-12
0.0
Lupus kidney
0.0

LAK cells IL-2 + IFN
0.0
NCI-H292 none
0.0

gamma

LAK cells IL-2 + IL-18
0.0
NCI-H292 IL-4
0.0

LAK cells
0.0
NCI-H292 IL-9
0.0

PMA/ionomycin

NK Cells IL-2 rest
0.0
NCI-H292 IL-13
0.0

Two Way MLR 3 day
0.0
NCI-H292 IFN gamma
0.0

Two Way MLR 5 day
0.0
HPAEC none
0.0

Two Way MLR 7 day
0.0
HPAEC TNF alpha + IL-1 beta
0.0

PBMC rest
0.0
Lung fibroblast none
0.0

PBMC PWM
0.0
Lung fibroblast TNF alpha + IL-
0.0

1 beta

PBMC PHA-L
0.0
Lung fibroblast IL-4
0.0

Ramos (B cell) none
0.0
Lung fibroblast IL-9
0.0

Ramos (B cell) ionomycin
0.0
Lung fibroblast IL-13
1.0

B lymphocytes PWM
0.0
Lung fibroblast IFN gamma
0.0

B lymphocytes CD40L
0.0
Dermal fibroblast CCD1070 rest
0.0

and IL-4

EOL-1 dbcAMP
0.0
Dermal fibroblast CCD1070
0.0

TNF alpha

EOL-1 dbcAMP
0.0
Dermal fibroblast CCD1070 IL-
0.0

PMA/ionomycin

1 beta

Dendritic cells none
0.0
Dermal fibroblast IFN gamma
0.0

Dendritic cells LPS
0.0
Dermal fibroblast IL-4
0.0

Dendritic cells anti-CD40
0.0
IBD Colitis 2
15.6

Monocytes rest
0.0
IBD Crohn's
4.2

Monocytes LPS
0.0
Colon
3.0

Macrophages rest
0.0
Lung
0.0

Macrophages LPS
0.0
Thymus
1.9

HUVEC none
0.0
Kidney
0.0

HUVEC starved
0.0

[0627] General_screening_panel_v1.5 Summary: Ag1834 Expression of the CG149828-01 gene is highest in a lung cancer cell line (CT=31.5). Significant expression of this gene is also detected in fetal skeletal muscle. Interestingly, this gene is expressed at much higher levels in fetal (CT=34.5) when compared to adult skeletal muscle (CT=40). This observation suggests that expression of this gene can be used to distinguish fetal from adult skeletal muscle. In addition, the relative overexpression of this gene in fetal skeletal muscle suggests that the protein product may enhance muscular growth or development in the fetus and thus may also act in a regenerative capacity in the adult. Therefore, therapeutic modulation of the GPCR encoded by this gene could be useful in treatment of muscle related diseases. More specifically, treatment of weak or dystrophic muscle with the protein encoded by this gene could restore muscle mass or function.

[0628] Panel 4D Summary: Ag1834 Expression of the CG149828-01 gene is highest in a liver cirrhosis sample. Furthermore, expression of this gene is not detected in normal liver in Panel 1.3D, suggesting that its expression is unique to liver cirrhosis. This gene encodes a putative GPCR; therefore, antibodies or small molecule therapeutics could reduce or inhibit fibrosis that occurs in liver cirrhosis. In addition, antibodies to this putative GPCR could also be used for the diagnosis of liver cirrhosis. In addition, this gene is also expressed at low levels in an IBD colitis sample.

[0629] W. GMAP002345_B: Olfactory Receptor

[0630] Expression of gene GMAP002345_B was assessed using the primer-probe sets Ag1728 and Ag1832, described in Tables WA and WB. Results of the RTQ-PCR runs are shown in Tables WC and WD.

105TABLE WAProbe Name Ag1728StartSEQ IDPrimersSequencesLengthPositionNO:Forward5′-tccatggacacagacaaaatg-3′21795265ProbeTET-5′-ccccatgctgaaccctctggtctata-3′-TAMRA26842266Reverse5′-cttcacttccttgttcctcaga-3′22869267

[0631]

106

TABLE WB

Probe Name Ag1832

Start
SEQ ID

Primers
Sequences
Length
Position
NO:

Forward
5′-tccatggacacagacaaaatg-3′
21
795
268

Probe
TET-5′-ccccatgctgaaccctctggtctata-3′-TAMRA
26
842
269

Reverse
5′-cttcacttccttgttcctcaga-3′
22
869
270

[0632]

107

TABLE WC

Panel 2.2

Rel. Exp. (%) Ag1728,

Rel. Exp. (%) Ag1728,

Tissue Name
Run 173761864
Tissue Name
Run 173761864

Normal Colon
0.0
Kidney Margin (OD04348)
0.0

Colon cancer (OD06064)
0.0
Kidney malignant cancer
0.0

(OD06204B)

Colon Margin (OD06064)
0.0
Kidney normal adjacent
0.0

tissue (OD06204E)

Colon cancer (OD06159)
0.0
Kidney Cancer (OD04450-
0.0

01)

Colon Margin (OD06159)
0.0
Kidney Margin (OD04450-
0.0

03)

Colon cancer (OD06297-04)
0.0
Kidney Cancer 8120613
0.0

Colon Margin (OD06297-
0.0
Kidney Margin 8120614
0.0

015)

CC Gr.2 ascend colon
0.0
Kidney Cancer 9010320
0.0

(ODO3921)

CC Margin (ODO3921)
0.0
Kidney Margin 9010321
0.0

Colon cancer metastasis
0.0
Kidney Cancer 8120607
0.0

(OD06104)

Lung Margin (OD06104)
0.0
Kidney Margin 8120608
0.0

Colon mets to lung
0.0
Normal Uterus
0.0

(OD04451-01)

Lung Margin (OD04451-02)
0.0
Uterine Cancer 064011
0.0

Normal Prostate
0.0
Normal Thyroid
0.0

Prostate Cancer (OD04410)
0.0
Thyroid Cancer 064010
0.0

Prostate Margin (OD04410)
0.0
Thyroid Cancer A302152
0.0

Normal Ovary
0.0
Thyroid Margin A302153
0.0

Ovarian cancer (OD06283-
0.0
Normal Breast
0.0

03)

Ovarian Margin (OD06283-
0.0
Breast Cancer (0D04566)
0.0

07)

Ovarian Cancer 064008
100.0
Breast Cancer 1024
0.0

Ovarian cancer (OD06145)
0.0
Breast Cancer (OD04590-
0.0

01)

Ovarian Margin (OD06145)
0.0
Breast Cancer Mets
0.0

(OD04590-03)

Ovarian cancer (OD06455-
0.0
Breast Cancer Metastasis
0.0

03)

(OD04655-05)

Ovarian Margin (OD06455-
0.0
Breast Cancer 064006
0.0

07)

Normal Lung
0.0
Breast Cancer 9100266
0.0

Invasive poor diff. lung
0.0
Breast Margin 9100265
0.0

adeno (ODO4945-01

Lung Margin (ODO4945-03)
0.0
Breast Cancer A209073
0.0

Lung Malignant Cancer
0.0
Breast Margin A2090734
0.0

(OD03126)

Lung Margin (OD03126)
0.0
Breast cancer (OD06083)
0.0

Lung Cancer (OD05014A)
0.0
Breast cancer node
0.0

metastasis (OD06083)

Lung Margin (OD05014B)
0.0
Normal Liver
0.0

Lung cancer (OD06081)
0.0
Liver Cancer 1026
0.0

Lung Margin (OD06081)
0.0
Liver Cancer 1025
9.2

Lung Cancer (OD04237-01)
0.0
Liver Cancer 6004-T
0.0

Lung Margin (OD04237-02)
0.0
Liver Tissue 6004-N
0.0

Ocular Melanoma Metastasis
0.0
Liver Cancer 6005-T
0.0

Ocular Melanoma Margin
0.0
Liver Tissue 6005-N
0.0

(Liver)

Melanoma Metastasis
0.0
Liver Cancer 064003
0.0

Melanoma Margin (Lung)
0.0
Normal Bladder
0.0

Normal Kidney
0.0
Bladder Cancer 1023
0.0

Kidney Ca, Nuclear grade 2
0.0
Bladder Cancer A302173
0.0

(OD04338)

Kidney Margin (OD04338)
0.0
Normal Stomach
0.0

Kidney Ca Nuclear grade 1/2
0.0
Gastric Cancer 9060397
0.0

(OD04339)

Kidney Margin (OD04339)
0.0
Stomach Margin 9060396
16.6

Kidney Ca, Clear cell type
0.0
Gastric Cancer 9060395
34.6

(OD04340)

Kidney Margin (OD04340)
0.0
Stomach Margin 9060394
0.0

Kidney Ca, Nuclear grade 3
0.0
Gastric Cancer 064005
0.0

(OD04348)

[0633]

108

TABLE WD

Panel 4D

Rel. Exp. (%)
Rel. Exp. (%)

Rel. Exp. (%)
Rel. Exp. (%)

Ag1728, Run
Ag1832, Run

Ag1728, Run
Ag1832, Run

Tissue Name
165364125
165810430
Tissue Name
165364125
165810430

Secondary Th1 act
0.0
0.0
HUVEC IL-1beta
0.0
0.0

Secondary Th2 act
0.0
0.0
HUVEC IFN gamma
0.0
0.0

Secondary Tr1 act
0.0
0.0
HUVEC TNF alpha +
0.0
2.1

IFN gamma

Secondary Th1 rest
0.0
0.0
HUVEC TNF alpha +
0.0
0.0

IL4

Secondary Th2 rest
0.0
0.0
HUVEC IL-11
0.0
0.0

Secondary Tr1 rest
0.0
0.0
Lung Microvascular
14.0
17.0

EC none

Primary Th1 act
0.0
0.0
Lung Microvascular
100.0
20.3

EC TNF alpha + IL-

1beta

Primary Th2 act
0.0
0.0
Microvascular
100.0
100.0

Dermal EC none

Primary Tr1 act
0.0
0.0
Microsvasular Dermal
0.0
8.1

EC TNF alpha + IL-

1beta

Primary Th1 rest
0.0
0.0
Bronchial epithelium
0.0
0.0

TNF alpha + IL1beta

Primary Th2 rest
13.5
0.0
Small airway
0.0
0.0

epithelium none

Primary Tr1 rest
0.0
0.0
Small airway
13.9
5.4

epithelium TNF alpha +

IL-1beta

CD45RA CD4
0.0
4.5
Coronery artery SMC
5.2
4.8

lymphocyte act

rest

CD45RO CD4
0.0
0.0
Coronery artery SMC
0.0
5.9

lymphocyte act

TNF alpha + IL-1beta

CD8 lymphocyte act
0.0
0.0
Astrocytes rest
0.0
0.0

Secondary CD8
0.0
0.0
Astrocytes TNF alpha +
0.0
11.3

lymphocyte rest

IL-1beta

Secondary CD8
0.0
4.5
KU-812 (Basophil)
0.0
0.0

lymphocyte act

rest

CD4 lymphocyte
0.0
0.0
KU-812 (Basophil)
0.0
0.0

none

PMA/ionomycin

2ry
0.0
0.0
CCD1106
0.0
1.0

Th1/Th2/Tr1_anti-

(Keratinocytes) none

CD95 CH11

LAK cells rest
0.0
0.0
CCD1106
0.0
1.1

(Keratinocytes)

TNF alpha + IL-1beta

LAK cells IL-2
0.0
0.0
Liver cirrhosis
88.9
71.7

LAK cells IL-2 + IL-
0.0
0.0
Lupus kidney
0.0
0.0

12

LAK cells IL-2 + IFN
0.0
0.0
NCI-H292 none
0.0
0.0

gamma

LAK cells IL-2 + IL-
0.0
0.0
NCI-H292 IL-4
0.0
0.0

18

LAK cells
0.0
0.0
NCI-H292 IL-9
3.2
0.0

PMA/ionomycin

NK Cells IL-2 rest
0.0
0.0
NCI-H292 IL-13
0.0
0.0

Two Way MLR 3
0.0
0.0
NCI-H292 IFN
0.0
0.0

day

gamma

Two Way MLR 5
0.0
0.0
HPAEC none
0.0
0.0

day

Two Way MLR 7
0.0
0.0
HPAEC TNF alpha +
0.0
0.0

day

IL-1beta

PBMC rest
0.0
0.0
Lung fibroblast none
0.0
2.5

PBMC PWM
13.6
0.0
Lung fibroblast TNF
0.0
0.0

alpha + IL-1beta

PBMC PHA-L
0.0
0.0
Lung fibroblast IL-4
0.0
0.0

Ramos (B cell) none
0.0
0.0
Lung fibroblast IL-9
0.0
0.0

Ramos (B cell)
0.0
0.0
Lung fibroblast IL-13
0.0
0.0

ionomycin

B lymphocytes PWM
0.0
0.0
Lung fibroblast IFN
0.0
0.0

gamma

B lymphocytes
0.0
0.0
Dermal fibroblast
0.0
0.0

CD40L and IL-4

CCD1070 rest

EOL-1 dbcAMP
0.0
0.0
Dermal fibroblast
0.0
0.0

CCD1070 TNF alpha

EOL-1 dbcAMP
0.0
0.0
Dermal fibroblast
0.0
0.0

PMA/ionomycin

CCD1070 IL-1beta

Dendritic cells none
0.0
0.0
Dermal fibroblast IFN
0.0
0.0

gamma

Dendritic cells LPS
0.0
6.3
Dermal fibroblast IL-4
0.0
0.0

Dendritic cells anti-
0.0
0.0
IBD Colitis 2
3.1
12.0

CD40

Monocytes rest
0.0
0.0
IBD Crohn's
0.0
0.0

Monocytes LPS
0.0
6.2
Colon
0.0
5.2

Macrophages rest
0.0
6.2
Lung
0.0
9.2

Macrophages LPS
0.0
0.0
Thymus
6.7
0.0

HUVEC none
0.0
0.0
Kidney
0.0
0.0

HUVEC starved
0.0
0.0

[0634] Panel 1.3D Summary: Ag1728 Expression of the GMAP002345_B gene is low/undetectable (CTs>35) across all of the samples on this panel (data not shown).

[0635] Panel 2.2 Summary: Ag1728 Significant expression of the GMAP002345_B gene is seen exclusively in an ovarian cancer sample (CT=34.5). Therefore, expression of this gene may be used to distinguish ovarian cancers from the other samples on this panel. Furthermore, therapeutic modulation of the activity of the GPCR encoded by this gene may be beneficial in the treatment of ovarian cancer.

[0636] Panel 4D Summary: Ag1728/1832 Two experiments with the same probe and primer set produce results that are in very good agreement, with highest expression in both runs in microvascular dermal endothelial cells and lung microvascular endothelial cells treated with TNFalpha+IL−1beta (CTs=3-34). Low but still significant levels of expression are also detected in samples from liver cirrhosis. Endothelial cells are known to play important roles in inflammatory responses by altering the expression of surface proteins that are involved in activation and recruitment of effector inflammatory cells. The expression of this gene in dermal microvascular endothelial cells suggests that this protein product may be involved in inflammatory responses to skin disorders, including psoriasis. Expression in lung microvascular endothelial cells suggests that the protein encoded by this transcript may also be involved in lung disorders including asthma, allergies, chronic obstructive pulmonary disease, and emphysema. Therefore, therapeutic modulation of the protein encoded by this gene may lead to amelioration of symptoms associated with psoriasis, asthma, allergies, chronic obstructive pulmonary disease, and emphysema. In addition, the expression in liver cirhoosis suggests that antibodies or small molecule therapeutics could reduce or inhibit fibrosis and other inflammatory processes that occur in liver cirrhosis. Furthermore, antibodies to this putative GPCR could also be used for the diagnosis of liver cirrhosis.

[0637] X. CG55962-01/GMAL391156_A: Olfactory Receptor

[0638] Expression of gene CG55962-01 was assessed using the primer-probe sets Ag1789 and Ag1714, described in Tables XA and XB. Results of the RTQ-PCR runs are shown in Table XC.

109TABLE XAProbe Name Ag1789StartSEQ IDPrimersSequencesLengthPositionNO:Forward5′-gtgggtaacagcctcatagtca-3′22121271ProbeTET-5′-tggaccctcacctacactctcctatg-3′-TAMRA26155272Reverse5′-tgaaagattggtaagcaggaaa-3′22183273

[0639]

110

TABLE XB

Probe Name Ag1714

Start
SEQ ID

Primers
Sequences
Length
Position
NO:

Forward
5′-gtgggtaacagcctcatagtca-3′
22
121
274

Probe
TET-5′-tggaccctcacctacactctcctatg-3′-TAMRA
26
155
275

Reverse
5′-tgaaagattggtaagcaggaaa-3′
22
183
276

[0640]

111

TABLE XC

Panel 4D

Rel. Exp. (%)
Rel. Exp. (%)

Rel. Exp. (%)
Rel. Exp. (%)

Ag1714, Run
Ag1789, Run

Ag1714, Run
Ag1789, Run

Tissue Name
165330745
165809174
Tissue Name
165330745
165809174

Secondary Th1 act
0.0
0.0
HUVEC IL-1beta
0.0
0.0

Secondary Th2 act
0.0
0.0
HUVEC IFN gamma
0.0
0.0

Secondary Tr1 act
0.0
0.0
HUVEC TNF alpha +
0.0
0.0

IFN gamma

Secondary Th1 rest
0.0
0.0
HUVEC TNF alpha +
0.0
0.0

IL4

Secondary Th2 rest
0.0
0.0
HUVEC IL-11
0.0
0.0

Secondary Tr1 rest
17.3
0.0
Lung Microvascular
0.0
0.0

EC none

Primary Th1 act
0.0
0.0
Lung Microvascular
0.0
0.0

EC TNF alpha + IL-

1beta

Primary Th2 act
27.9
0.0
Microvascular
0.0
0.0

Dermal EC none

Primary Tr1 act
0.0
0.0
Microsvasular Dermal
0.0
0.0

EC TNF alpha + IL-

1beta

Primary Th1 rest
0.0
0.0
Bronchial epithelium
0.0
0.0

TNF alpha + IL1beta

Primary Th2 rest
0.0
0.0
Small airway
0.0
0.0

epithelium none

Primary Tr1 rest
0.0
0.0
Small airway
0.0
0.0

epithelium TNF alpha +

IL-1beta

CD45RA CD4
0.0
0.0
Coronery artery SMC
0.0
0.0

lymphocyte act

rest

CD45RO CD4
0.0
0.0
Coronery artery SMC
0.0
0.0

lymphocyte act

TNF alpha + IL-1beta

CD8 lymphocyte act
0.0
0.0
Astrocytes rest
0.0
0.0

Secondary CD8
0.0
0.0
Astrocytes TNF alpha +
0.0
0.0

lymphocyte rest

IL-1beta

Secondary CD8
0.0
0.0
KU-812 (Basophil)
0.0
0.0

lymphocyte act

rest

CD4 lymphocyte
0.0
0.0
KU-812 (Basophil)
0.0
0.0

none

PMA/ionomycin

2ry
7.0
0.0
CCD1106
0.0
0.0

Th1/Th2/Tr1_anti-

(Keratinocytes) none

CD95 CH11

LAK cells rest
0.0
0.0
CCD1106
0.0
0.0

(Keratinocytes)

TNF alpha + IL-1beta

LAK cells IL-2
0.0
0.0
Liver cirrhosis
100.0
100.0

LAK cells IL-2 + IL-
0.0
0.0
Lupus kidney
0.0
0.0

12

LAK cells IL-2 + IFN
0.0
0.0
NCI-H292 none
0.0
0.0

gamma

LAK cells IL-2 + IL-
0.0
0.0
NCI-H292 IL-4
0.0
0.0

18

LAK cells
0.0
0.0
NCI-H292 IL-9
0.0
0.0

PMA/ionomycin

NK Cells IL-2 rest
0.0
0.0
NCI-H292 IL-13
0.0
0.0

Two Way MLR 3
0.0
0.0
NCI-H292 IFN
0.0
0.0

day

gamma

Two Way MLR 5
0.0
0.0
HPAEC none
0.0
0.0

day

Two Way MLR 7
0.0
0.0
HPAEC TNF alpha +
0.0
0.0

day

IL-1beta

PBMC rest
0.0
0.0
Lung fibroblast none
0.0
0.0

PBMC PWM
0.0
0.0
Lung fibroblast TNF
7.9
0.0

alpha + IL-1beta

PBMC PHA-L
0.0
0.0
Lung fibroblast IL-4
0.0
0.0

Ramos (B cell) none
0.0
0.0
Lung fibroblast IL-9
0.0
0.0

Ramos (B cell)
0.0
0.0
Lung fibroblast IL-13
0.0
0.0

ionomycin

B lymphocytes PWM
0.0
0.0
Lung fibroblast IFN
0.0
0.0

gamma

B lymphocytes
0.0
0.0
Dermal fibroblast
0.0
0.0

CD40L and IL-4

CCD-1070 rest

EOL-1 dbcAMP
0.0
0.0
Dermal fibroblast
0.0
0.0

PMA/ionomycin

CCD1070 IL-1beta

EOL-1 dbcAMP
0.0
0.0
Dermal fibroblast
0.0
0.0

CCD1070 IL-1beta

Dendritic cells none
0.0
0.0
Dermal fibroblast IFN
0.0
0.0

gamma

Dendritic cells LPS
0.0
0.0
Dermal fibroblast IL-4
38.2
0.0

Dendritic cells anti-
0.0
0.0
IBD Colitis 2
60.3
51.8

CD40

Monocytes rest
0.0
0.0
IBD Crohn's
55.9
10.4

Monocytes LPS
0.0
2.1
Colon
0.0
3.7

Macrophages rest
22.5
0.0
Lung
0.0
0.0

Macrophages LPS
0.0
0.0
Thymus
0.0
1.4

HUVEC none
0.0
0.0
Kidney
0.0
1.6

HUVEC starved
0.0
0.0

[0641] Panel 1.3D Summary: Ag1789 Expression of the CG55962-01 gene is low/undetectable (CTs>35) across all of the samples on this panel (data not shown).

[0642] Panel 2.2 Summary: Ag1789 Expression of the CG55962-01 gene is low/undetectable (CTs>35) across all of the samples on this panel (data not shown).

[0643] Panel 4D Summary: Ag1714/Ag1789 Results from two experiments using an identical probe/primer set show reasonable agreement. Significant expression of the CG55962-01 gene is detected in a liver cirrhosis sample. Furthermore, expression of this gene is not detected in normal liver in Panel 1.3D, suggesting that its expression is unique to liver cirrhosis. This gene encodes a putative GPCR; therefore, antibodies or small molecule therapeutics could reduce or inhibit fibrosis that occurs in liver cirrhosis. In addition, antibodies to this putative GPCR could also be used for the diagnosis of liver cirrhosis (Mark et al., G protein modulation of recombinant P/Q-type calcium channels by regulators of G protein signalling proteins. J. Physiol. 528 Pt 1:65-77, 2000).

[0644] Y. CG55544-03/GMAC019093_A: Olfactory Receptor

[0645] Expression of gene CG55544-03 was assessed using the primer-probe sets Ag1234 and Ag1708, described in Tables YA and YB. Results of the RTQ-PCR runs are shown in Tables YC, YD, and YE.

112TABLE YAProbe Name Ag1234StartSEQ IDPrimersSequencesLengthPositionNO:Forward5′-tggaccttctgtctgaaagaaa-3′22257277ProbeTET-5′-ccatctccttcaatcattgcttcactca-3′-TAMRA28281278Reverse5′-atacatccacccctccaataag-3′22325279

[0646]

113

TABLE YB

Probe Name Ag1708

Start
SEQ ID

Primers
Sequences
Length
Position
NO:

Forward
5′-tggaccttctgtctgaaagaaa-3′
22
257
280

Probe
TET-5′-ccatctccttcaatcattgcttcactca-3′-TAMRA
28
281
281

Reverse
5′-atacatccacccctccaataag-3′
22
325
282

[0647]

114

TABLE YC

Panel 1.3D

Rel. Exp. (%) Ag1708, Run

Rel. Exp. (%) Ag1708, Run

Tissue Name
165925623
Tissue Name
165925623

Liver adenocarcinoma
0.0
Kidney (fetal)
0.0

Pancreas
0.0
Renal ca. 786-0
0.0

Pancreatic ca. CAPAN 2
0.0
Renal ca. A498
0.0

Adrenal gland
0.0
Renal ca. RXF 393
0.0

Thyroid
0.0
Renal ca. ACHN
0.0

Salivary gland
0.0
Renal ca. UO-31
0.0

Pituitary gland
0.0
Renal ca. TK-10
0.0

Brain (fetal)
0.0
Liver
0.0

Brain (whole)
0.0
Liver (fetal)
0.0

Brain (amygdala)
0.0
Liver ca. (hepatoblast)
0.0

HepG2

Brain (cerebellum)
0.0
Lung
0.0

Brain (hippocampus)
0.0
Lung (fetal)
0.0

Brain (substantia nigra)
0.0
Lung ca. (small cell) LX-1
0.0

Brain (thalamus)
0.0
Lung ca. (small cell)
0.0

NCI-H69

Cerebral Cortex
0.0
Lung ca. (s.cell var.)
0.0

SHP-77

Spinal cord
0.0
Lung ca. (large cell)NCI-
0.0

H460

glio/astro U87-MG
0.0
Lung ca. (non-sm. cell)
0.0

A549

glio/astro U-118-MG
0.0
Lung ca. (non-s.cell)
0.0

NCI-H23

astrocytoma SW1783
0.0
Lung ca. (non-s.cell)
0.0

HOP-62

neuro*; met SK-N-AS
0.0
Lung ca. (non-s.cl) NCI-
0.0

H522

astrocytoma SF-539
0.0
Lung ca. (squam.) SW
0.0

900

astrocytoma SNB-75
0.0
Lung ca. (squam.) NCI-
0.0

H596

glioma SNB-19
6.9
Mammary gland
0.0

glioma U251
0.0
Breast ca.* (pl.ef) MCF-7
0.0

glioma SF-295
0.0
Breast ca.* (pl.ef) MDA-
0.0

MB-231

Heart (Fetal)
0.0
Breast ca.* (pl. ef) T47D
0.0

Heart
0.0
Breast ca. BT-549
0.0

Skeletal muscle (Fetal)
0.0
Breast ca. MDA-N
0.0

Skeletal muscle
0.0
Ovary
7.5

Bone marrow
0.0
Ovarian ca. OVCAR-3
0.0

Thymus
0.0
Ovarian ca. OVCAR-4
0.0

Spleen
100.0
Ovarian ca. OVCAR-5
0.0

Lymph node
0.0
Ovarian ca. OVCAR-8
0.0

Colorectal
0.0
Ovarian ca. IGROV-1
0.0

Stomach
0.0
Ovarian ca. (ascites) SK-
8.9

OV-3

Small intestine
0.0
Uterus
0.0

Colon ca. SW480
0.0
Placenta
0.0

Colon ca.* SW620
0.0
Prostate
0.0

(SW480 met)

Colon ca. HT29
0.0
Prostate ca.* (bone met)
0.0

PC-3

Colon ca. HCT-116
0.0
Testis
0.0

Colon ca. CaCo-2
0.0
Melanoma Hs688(A).T
0.0

CC Well to Mod Diff
0.0
Melanoma* (met)
0.0

(ODO3866)

Hs688(B).T

Colon ca. HCC-2998
0.0
Melanoma UACC-62
0.0

Gastric ca. (liver met)
0.0
Melanoma M14
0.0

NCI-N87

Bladder
0.0
Melanoma LOX IMVI
0.0

Trachea
0.0
Melanoma* (met) SK-
12.9

MEL-5

Kidney
0.0
Adipose
0.0

[0648]

115

TABLE YE

Panel 2.2

Rel. Exp. (%) Ag1708,

Rel. Exp. (%) Ag1708,

Tissue Name
Run 173761450
Tissue Name
Run 173761450

Normal Colon
0.0
Kidney Margin (OD04348)
0.0

Colon cancer (OD06064)
0.0
Kidney malignant cancer
0.0

(OD06204B)

Colon Margin (OD06064)
0.0
Kidney normal adjacent
0.0

tissue (OD06204E)

Colon cancer (OD06159)
0.0
Kidney Cancer (OD04450-
0.0

01)

Colon Margin (OD06159)
0.0
Kidney Margin (OD04450-
0.0

03)

Colon cancer (OD06297-04)
0.0
Kidney Cancer 8120613
0.0

Colon Margin (OD06297-
0.0
Kidney Margin 8120614
0.0

015)

CC Gr.2 ascend colon
0.0
Kidney Cancer 9010320
0.0

(ODO3921)

CC Margin (ODO3921)
0.0
Kidney Margin 9010321
0.0

Colon cancer metastasis
0.0
Kidney Cancer 8120607
0.0

(OD06104)

Lung Margin (OD06104)
0.0
Kidney Margin 8120608
0.0

Colon mets to lung
0.0
Normal Uterus
0.0

(OD04451-01)

Lung Margin (OD04451-02)
0.0
Uterine Cancer 064011
0.0

Normal Prostate
0.0
Normal Thyroid
0.0

Prostate Cancer (OD04410)
0.0
Thyroid Cancer
0.0

Prostate Margin (OD04410)
0.0
Thyroid Cancer A302152
100.0

Normal Ovary
0.0
Thyroid Margin A302153
0.0

Ovarian cancer (OD06283-
0.0
Normal Breast
0.0

03)

Ovarian Margin (OD06283-
0.0
Breast Cancer
16.6

07)

Ovarian Cancer
59.9
Breast Cancer
0.0

Ovarian cancer (OD06145)
0.0
Breast Cancer (OD04590-
0.0

01)

Ovarian Margin (OD06145)
5.5
Breast Cancer Mets
0.0

(OD04590-03)

Ovarian cancer (OD06455-
0.0
Breast Cancer Metastasis
0.0

03)

Ovarian Margin (OD06455-
0.0
Breast Cancer
0.0

07

Normal Lung
0.0
Breast Cancer 9100266
0.0

Invasive poor diff. lung
0.0
Breast Margin 9100265
0.0

adeno (ODO4945-01

Lung Margin (ODO4945-03)
0.0
Breast Cancer A209073
0.0

Lung Malignant Cancer
0.0
Breast Margin A2090734
0.0

(OD03126)

Lung Margin (OD03126)
0.0
Breast cancer (OD06083)
8.0

Lung Cancer (OD05014A)
0.0
Breast cancer node
0.0

metastasis (OD06083)

Lung Margin (OD05014B)
0.0
Normal Liver
0.0

Lung cancer (OD06081)
0.0
Liver Cancer 1026
0.0

Lung Margin (OD06081)
0.0
Liver Cancer 1025
2.9

Lung Cancer (OD04237-01)
0.0
Liver Cancer 6004-T
0.0

Lung Margin (OD04237-02)
0.0
Liver Tissue 6004-N
0.0

Ocular Mel Met to Liver
0.0
Liver Cancer 6005-T
0.0

(ODO4310)

Liver Margin (ODO4310)
0.0
Liver Tissue 6005-N
2.4

Melanoma Metastasis
0.0
Liver Cancer
0.0

Lung Margin (OD04321)
0.0
Normal Bladder
0.0

Normal Kidney
0.0
Bladder Cancer
0.0

Kidney Ca, Nuclear grade 2
0.0
Bladder Cancer
0.0

(OD04338)

Kidney Margin (OD04338)
0.0
Normal Stomach
0.0

Kidney Ca Nuclear grade 1/2
0.0
Gastric Cancer 9060397
0.0

(OD04339)

Kidney Margin (OD04339)
0.0
Stomach Margin 9060396
12.6

Kidney Ca, Clear cell type
0.0
Gastric Cancer 9060395
2.7

(OD04340)

Kidney Margin (OD04340)
0.0
Stomach Margin 9060394
0.0

Kidney Ca, Nuclear grade 3
0.0
Gastric Cancer 064005
0.0

(OD04348)

[0649]

116

TABLE YF

Panel 4D

Rel. Exp. (%) Ag1708,

Rel. Exp. (%) Ag1708,

Tissue Name
Run 165813791
Tissue Name
Run 165813791

Secondary Th1 act
0.0
HUVEC IL-1 beta
0.0

Secondary Th2 act
0.0
HUVEC IFN gamma
0.0

Secondary Tr1 act
3.6
HUVEC TNF alpha + IFN
0.0

gamma

Secondary Th1 rest
0.0
HUVEC TNF alpha + IL4
0.0

Secondary Th2 rest
0.0
HUVEC IL-11
0.0

Secondary Tr1 rest
0.0
Lung Microvascular EC none
0.0

Primary Th1 act
0.0
Lung Microvascular EC
0.0

TNF alpha + IL-1 beta

Primary Th2 act
0.0
Microvascular Dermal EC none
0.0

Primary Tr1 act
0.0
Microsvasular Dermal EC
0.0

TNF alpha + IL-1 beta

Primary Th1 rest
0.0
Bronchial epithelium TNF alpha +
0.0

IL-1 beta

Primary Th2 rest
0.0
Small airway epithelium none
0.0

Primary Tr1 rest
0.0
Small airway epithelium
0.0

TNF alpha + IL-1 beta

CD45RA CD4 lymphocyte
0.0
Coronery artery SMC rest
0.0

act

CD45RO CD4 lymphocyte
0.0
Coronery artery SMC TNF alpha +
0.0

act

IL-1 beta

CD8 lymphocyte act
0.0
Astrocytes rest
0.0

Secondary CD8
0.0
Astrocytes TNF alpha + IL-1 beta
0.0

lymphocyte rest

Secondary CD8
0.0
KU-812 (Basophil) rest
0.0

lymphocyte act

CD4 lymphocyte none
0.0
KU-812 (Basophil)
0.0

PMA/ionomycin

2ry Th1/Th2/Tr1_anti-
0.0
CD1106 (Keratinocytes) none
0.0

CD95 CH11

LAK cells rest
0.0
CCD1106 (Keratinocytes)
0.0

TNF alpha + IL-1 beta

LAK cells IL-2
0.0
Liver cirrhosis
100.0

LAK cells IL-2 + IL-12
0.0
Lupus kidney
0.0

LAK cells IL-2 + IFN
0.0
NCI-H292 none
0.0

gamma

LAK cells IL-2 + IL-18
0.0
NCI-H292 IL-4
0.0

LAK cells
0.0
NCI-H292 IL-9
0.0

PMA/ionomycin

NK Cells IL-2 rest
0.0
NCI-H292 IL-13
0.0

Two Way MLR 3 day
0.0
NCI-H292 IFN gamma
0.0

Two Way MLR 5 day
0.0
HPAEC none
0.0

Two Way MLR 7 day
0.0
HPAEC TNF alpha + IL-1 beta
0.0

PBMC rest
0.0
Lung fibroblast none
0.0

PBMC PWM
0.0
Lung fibroblast TNF alpha + IL-
0.0

1 beta

PBMC PHA-L
0.0
Lung fibroblast IL-4
0.0

Ramos (B cell) none
0.0
Lung fibroblast IL-9
0.0

Ramos (B cell) ionomycin
0.0
Lung fibroblast IL-13
0.0

B lymphocytes PWM
0.0
Lung fibroblast IFN gamma
0.0

B lymphocytes CD40L
0.0
Dermal fibroblast CCD1070 rest
0.0

and IL-4

EOL-1 dbcAMP
0.0
Dermal fibroblast CCD1070
0.0

TNF alpha

EOL-1 dbcAMP
5.3
Dermal fibroblast CCD1070 IL-
0.0

PMA/ionomycin

1 beta

Dendritic cells none
0.0
Dermal fibroblast IFN gamma
0.0

Dendritic cells LPS
0.0
Dermal fibroblast IL-4
0.0

Dendritic cells anti-CD40
0.0
IBD Colitis 2
6.4

Monocytes rest
0.0
IBD Crohn's
0.0

Monocytes LPS
0.0
Colon
7.1

Macrophages rest
0.0
Lung
0.0

Macrophages LPS
0.0
Thymus
0.0

HUVEC none
0.0
Kidney
0.0

HUVEC starved
0.0

[0650] Panel 1.2 Summary: Ag1234 Expression of the CG55544-03 gene is low/undetectable (CTs>35) across all of the samples on this panel (data not shown).

[0651] Panel 1.3D Summary: Ag1708 Significant expression of the CG55544-03 gene is restricted to the spleen, an important site of secondary immune responses (CT=34.6). Therefore, expression of this gene in spleen can be used to distinguish spleen from the other samples on this panel. Furthermore, antibodies or small molecule therapeutics that block the function of this GPCR may be useful as anti-inflammatory therapeutics for the treatment of allergies, autoimmune diseases, and inflammatory diseases.

[0652] Panel 2.2 Summary: Ag1708 The CG55544-03 gene is most highly expressed in a thyroid cancer sample (CT=33.5). Interestingly, levels of expression of this gene in the thyroid tumor are much higher than in the normal adjacent tissue. Therefore, expression of this gene may be used as a marker to distinguish normal thyroid from thyroid tumors. Furthermore, therapeutic modulation of the activity of the GPCR encoded by this gene may be beneficial in the treatment of thyroid cancer. Low but significant expression of this gene is also seen in an ovarian cancer sample.

[0653] Panel 4D Summary: Ag1708 Significant expression of the CG55544-03 gene is detected in a liver cirrhosis sample (CT=33.2). Furthermore, expression of this gene is not detected in normal liver in Panel 1.3D, suggesting that its expression is unique to liver cirrhosis. This gene encodes a putative GPCR; therefore, antibodies or small molecule therapeutics could reduce or inhibit fibrosis that occurs in liver cirrhosis. In addition, antibodies to this putative GPCR could also be used for the diagnosis of liver cirrhosis. Ag1234 Expression of this gene is low/undetectable (CTs>35) across all of the samples on this panel (data not shown).

[0654] Z. CG55769-01/GMAC022207_A and CG55769-02: Olfactory Receptor

[0655] Expression of gene CG55769-01 and variant CG55769-02 was assessed using the primer-probe sets Ag2596 and Ag1736, described in Tables ZA and ZB. Results of the RTQ-PCR runs are shown in Tables ZC, ZD and ZE.

117TABLE ZAProbe Name Ag2596StartSEQ IDPrimersSequencesLengthPositionNO:Forward5′-tcattgtggtgtccctcttcta-3′22736283ProbeTET-5′-cggaacagccagtctgacctacctg-3′-TAMRA25758284Reverse5′-tgacactagcttcttgctctca-3′22806285

[0656]

118

TABLE ZB

Probe Name Ag1736

Start
SEQ ID

Primers
Sequences
Length
Position
NO:

Foward
5′-cacccgattatcataacaatt-3′
22
656
286

Probe
TET-5′-actggccgccagaaggcattttcta-3′-TAMRA
25
696
287

Reverse
5′-acaatgaggtgtgaggaacaag-3′
22
721
288

[0657]

119

TABLE ZC

Panel 1.3D

Rel. Exp. (%)
Rel. Exp. (%)

Rel. Exp. (%)
Rel. Exp. (%)

Ag1736, Run
Ag2596, Run

Ag1736, Run
Ag2596, Run

Tissue Name
165940873
165645336
Tissue Name
165940873
165645336

Liver
70.7
4.7
Kidney (fetal)
0.0
0.0

adenocarcinoma

Pancreas
0.0
6.1
Renal ca. 786-0
0.0
0.0

Pancreatic ca.
0.0
0.0
Renal ca. A498
0.0
0.0

CAPAN2

Adrenal gland
0.0
0.0
Renal ca. RXF
0.0
3.6

393

Thyroid
0.0
0.0
Renal ca. ACHN
0.0
0.0

Salivary gland
6.8
0.0
Renal ca. UO-31
0.0
0.0

Pituitary gland
0.0
0.0
Renal ca. TK-10
0.0
0.0

Brain (fetal)
0.0
0.0
Liver
0.0
10.4

Brain (whole)
0.0
8.7
Liver (fetal)
0.0
0.0

Brain (amygdala)
0.0
5.3
Liver ca.
0.0
0.0

(hepatoblast)

HepG2

Brain (cerebellum)
0.0
0.0
Lung
0.0
0.0

Brain
0.0
0.0
Lung (fetal)
0.0
0.0

(hippocampus)

Brain (substantia
0.0
5.7
Lung ca. (small
0.0
0.0

nigra)

cell) LX-1

Brain (thalamus)
0.0
0.0
Lung ca. (small
0.0
0.0

cell) NCI-H69

Cerebral Cortex
0.0
0.0
Lung ca. (s. cell
0.0
0.0

var.) SHP-77

Spinal cord
0.0
0.0
Lung ca. (large
0.0
7.4

cell)NCI-H460

glio/astro U87-MG
0.0
0.0
Lung ca. (non-
0.0
0.0

sm. cell) A549

glio/astro U-118-
0.0
0.0
Lung ca. (non-
0.0
1.4

MG

s. cell) NCI-H23

astrocytoma
0.0
1.9
Lung ca. (non-
0.0
2.5

SW1783

s. cell) HOP-62

neuro*; met SK-N
0.0
0.0
Lung ca. (non-
0.0
0.0

AS

s. cl) NCI-H522

astrocytoma SF-539
0.0
0.0
Lung ca.
0.0
0.0

(squam.) SW 900

astrocytoma SNB-
0.0
0.0
Lung ca.
0.0
0.3

75

(squam.) NCI-

H596

glioma SNB-19
0.0
0.0
Mammary gland
0.0
11.2

glioma U251
0.0
0.0
Breast ca.*
0.0
0.0

(pl. ef) MCF-7

glioma SF-295
0.0
0.0
Breast ca.*
0.0
0.0

(pl. ef) MDA-

MB-231

Heart (Fetal)
0.0
0.0
Breast ca.* (pl.
0.0
0.0

ef) T47D

Heart
0.0
0.0
Breast ca. BT-
0.0
5.1

549

Skeletal muscle
0.0
0.0
Breast ca. MDA-N
0.0
0.0

(Fetal)

Skeletal muscle
0.0
0.0
Ovary
0.0
0.0

Bone marrow
0.0
0.0
Ovarian ca.
0.0
0.0

OVCAR-3

Thymus
0.0
4.5
Ovarian ca.
0.0
0.0

OVCAR-4

Spleen
100.0
0.0
Ovarian ca.
0.0
0.0

OVCAR-5

Lymph node
0.0
0.0
Ovarian ca.
0.0
0.0

OVCAR-8

Colorectal
0.0
0.0
Ovarian ca.
0.0
0.0

IGROV-1

Stomach
0.0
2.4
Ovarian ca.
16.5
3.2

(ascites) SK-OV-3

Small intestine
0.0
0.0
Uterus
0.0
0.0

Colon ca. SW480
0.0
0.0
Placenta
0.0
0.0

Colon ca.* SW620
0.0
0.0
Prostate
0.0
2.5

(SW480 met)

Colon ca. HT29
0.0
100.0
Prostate ca.*
0.0
0.0

(bone met) PC-3

Colon ca. HCT-116
0.0
0.0
Testis
0.0
0.0

Colon ca. CaCo-2
10.7
0.0
Melanoma
0.0
0.0

Hs688(A).T

CC Well to Mod
0.0
2.1
Melanoma*
0.0
0.0

Diff (ODO3866)

(met)

Hs688(B).T

Colon ca. HCC-
0.0
5.3
Melanoma
0.0
0.0

2998

UACC-62

Gastric ca. (liver
0.0
0.0
Melanoma M14
0.0
0.0

met) NCI-N87

Bladder
0.0
9.9
Melanoma LOX
0.0
0.0

IMVI

Trachea
0.0
0.0
Melanoma*
0.0
0.0

(met SK-MEL-5

Kidney
0.0
3.1
Adipose
0.0
0.0

[0658]

120

TABLE ZD

Panel 2.2

Rel. Exp. (%)
Rel. Exp. (%)

Rel. Exp. (%)
Rel. Exp. (%)

Ag1736, Run
Ag2596, Run

Ag1736, Run
Ag2596, Run

Tissue Name
174111847
175127796
Tissue Name
174111847
175127796

Normal Colon
0.0
0.0
Kidney Margin
0.0
0.0

(OD04348)

Colon cancer
0.0
0.0
Kidney malignant
60.7
21.5

(OD06064)

cancer

(OD06204B)

Colon Margin
0.0
0.0
Kidney normal
0.0
0.0

(OD06064)

adjacent tissue

(OD06204E)

Colon cancer
0.0
0.0
Kidney Cancer
0.0
0.0

(OD06159)

(OD04450-01)

Colon Margin
0.0
0.0
Kidney Margin
0.0
0.0

(OD06159)

(OD04450-03)

Colon cancer
0.0
0.0
Kidney Cancer
0.0
0.0

(OD06297-04)

8120613

Colon Margin
0.0
0.0
Kidney Margin
0.0
0.0

(OD06297-015)

8120614

CC Gr.2 ascend
0.0
0.0
Kidney Cancer
0.0
0.0

colon (ODO3921)

9010320

CC Margin
0.0
0.0
Kidney Margin
0.0
0.0

(ODO3921)

9010321

Colon cancer
0.0
0.0
Kidney Cancer
0.0
0.0

metastasis

8120607

(OD06104)

Lung Margin
0.0
0.0
Kidney Margin
0.0
0.0

(OD06104)

8120608

Colon mets to lung
0.0
0.0
Normal Uterus
0.0
0.0

(OD04451-01)

Lung Margin
0.0
0.0
Uterine Cancer
0.0
0.0

(OD04451-02)

064011

Normal Prostate
0.0
0.0
Normal Thyroid
0.0
0.0

Prostate Cancer
0.0
0.0
Thyroid Cancer
0.0
0.0

(OD04410)

Prostate Margin
0.0
0.0
Thyroid Cancer
0.0
0.0

(OD04410)

A302152

Normal Ovary
0.0
0.0
Thyroid Margin
0.0
0.0

A302153

Ovarian cancer
0.0
0.0
Normal Breast
0.0
0.0

(OD06283-03)

Ovarian Margin
0.0
0.0
Breast Cancer
0.0
0.0

(OD06283-07)

Ovarian Cancer
0.0
20.7
Breast Cancer
0.0
28.7

Ovarian cancer
0.0
0.0
Breast Cancer
0.0
0.0

(OD06145)

(OD04590-01)

Ovarian Margin
0.0
0.0
Breast Cancer Mets
0.0
0.0

(OD06145)

(OD04590-03)

Ovarian cancer
0.0
0.0
Breast Cancer
100.0
100.0

(OD06455-03)

Metastasis

Ovarian Margin
0.0
0.0
Breast Cancer
0.0
0.0

(OD06455-07)

Normal Lung
0.0
0.0
Breast Cancer
0.0
0.0

9100266

Invasive poor diff.
0.0
0.0
Breast Margin
43.8
0.0

lung adeno

9100265

(ODO4945-01

Lung Margin
0.0
13.0
Breast Cancer
0.0
0.0

(ODO4945-03)

A209073

Lung Malignant
0.0
0.0
Breast Margin
38.7
0.0

Cancer (OD03126)

A2090734

Lung Margin
0.0
0.0
Breast cancer
0.0
0.0

(OD03126)

(OD06083)

Lung Cancer
0.0
0.0
Breast cancer node
0.0
0.0

(OD05014A)

metastasis

(OD06083)

Lung Margin
0.0
0.0
Normal Liver
0.0
0.0

(OD05014B)

Lung cancer
0.0
0.0
Liver Cancer 1026
0.0
0.0

(OD06081)

Lung Margin
0.0
0.0
Liver Cancer 1025
0.0
0.0

(OD06081)

Lung Cancer
0.0
0.0
Liver Cancer 6004-T
0.0
0.0

(OD04237-01)

Lung Margin
0.0
0.0
Liver Tissue 6004-N
0.0
0.0

(OD04237-02)

Ocular Mel Met to
0.0
0.0
Liver Cancer 6005-T
0.0
0.0

Liver (ODO4310)

Liver Margin
0.0
0.0
Liver Tissue 6005-N
0.0
0.0

(ODO4310)

Melanoma
0.0
0.0
Liver Cancer
0.0
0.0

Metastasis

Lung Margin
0.0
0.0
Normal Bladder
0.0
0.0

(OD04321)

Normal Kidney
0.0
0.0
Bladder Cancer
0.0
0.0

Kidney Ca, Nuclear
0.0
0.0
Bladder Cancer
0.0
0.0

grade 2 (OD04338)

Kidney Margin
0.0
0.0
Normal Stomach
0.0
0.0

(OD04338)

Kidney Ca Nuclear
0.0
0.0
Gastric Cancer
0.0
0.0

grade 1/2

9060397

(OD04339)

Kidney Margin
0.0
0.0
Stomach Margin
0.0
0.0

(OD04339)

9060396

Kidney Ca, Clear
0.0
0.0
Gastric Cancer
52.9
26.1

cell type

9060395

(OD04340)

Kidney Margin
0.0
0.0
Stomach Margin
0.0
0.0

(OD04340)

9060394

Kidney Ca, Nuclear
0.0
0.0
Gastric Cancer
0.0
0.0

grade 3 (OD04348)

064005

[0659]

121

TABLE ZE

Panel 4D

Rel. Exp. (%)
Rel. Exp. (%)

Rel. Exp. (%)
Rel. Exp. (%)

Ag1736, Run
Ag2596, Run

Ag1736, Run
Ag2596, Run

Tissue Name
165813056
164395444
Tissue Name
165813056
164395444

Secondary Th1 act
0.0
0.0
HUVEC IL-1beta
0.0
0.0

Secondary Th2 act
3.7
0.0
HUVEC IFN gamma
0.0
0.0

Secondary Tr1 act
0.0
0.0
HUVEC TNF alpha +
0.0
0.0

IFN gamma

Secondary Th1 rest
0.0
0.0
HUVEC TNF alpha +
0.0
0.0

IL4

Secondary Th2 rest
0.0
0.0
HUVEC IL-11
0.0
0.0

Secondary Tr1 rest
0.0
0.0
Lung Microvascular
3.2
0.0

EC none

Primary Th1 act
0.0
0.0
Lung Microvascular
0.0
0.0

EC TNF alpha + IL-

1beta

Primary Th2 act
0.0
0.0
Microvascular
0.0
0.0

Dermal EC none

Primary Tr1 act
0.0
0.0
Microsvascular Dermal
0.0
0.0

EC TNF alpha + IL-

1beta

Primary Th1 rest
0.0
0.0
Bronchial epithelium
0.0
0.0

TNF alpha + IL 1beta

Primary Th2 rest
4.5
0.0
Small airway
0.0
0.0

epithelium none

Primary Tr1 rest
0.0
0.0
Small airway
0.0
0.0

epithelium TNF alpha +

IL-1beta

CD45RA CD4
0.0
0.0
Coronery artery SMC
0.0
0.0

lymphocyte act

rest

CD45RO CD4
0.0
0.0
Coronery artery SMC
0.0
0.0

lymphocyte act

TNF alpha + IL-1beta

CD8 lymphocyte act
0.0
0.0
Astrocytes rest
0.0
0.0

Secondary CD8
0.0
0.0
Astrocytes TNF alpha +
0.0
0.0

lymphocyte rest

IL-1beta

Secondary CD8
0.0
0.0
KU-812 (Basophil)
0.0
65.1

lymphocyte act

rest

CD4 lymphocyte
0.0
0.0
KU-812 (Basophil)
0.0
0.0

none

PMA/ionomycin

2ry
0.0
0.0
CCD1106
0.0
0.0

Th1/Th2/Tr1_anti-

(Keratinocytes) none

CD95 CH11

LAK cells rest
0.0
0.0
CCD1106
0.0
0.0

(Keratinocytes)

TNF alpha + IL-1beta

LAK cells IL-2
0.0
0.0
Liver cirrhosis
100.0
100.0

LAK cells IL-2 + IL-
0.0
0.0
Lupus kidney
0.0
0.0

12

LAK cells IL-2 + IFN
0.0
0.0
NCI-H292 none
0.0
0.0

gamma

LAK cells IL-2 + IL-
0.0
0.0
NCI-H292 IL-4
0.0
0.0

18

LAK cells
0.0
0.0
NCI-H292 IL-9
0.0
0.0

PMA/ionomycin

NK Cells IL-2 rest
0.0
0.0
NCI-H292 IL-13
0.0
0.0

Two Way MLR 3
0.0
0.0
NCI-H292 IFN
0.0
0.0

day

gamma

Two Way MLR 5
0.0
0.0
HPAEC none
0.0
0 0

day

Two Way MLR 7
0.0
0.0
HPAEC TNF alpha +
0.0
0.0

day

IL-1beta

PBMC rest
0.0
0.0
Lung fibroblast none
0.0
0.0

PBMC PWM
0.0
0.0
Lung fibroblast TNF
0.0
0.0

alpha + IL-1beta

PBMC PHA-L
0.0
28.3
Lung fibroblast IL-4
0.0
0.0

Ramos (B cell) none
0.0
0.0
Lung fibroblast IL-9
0.0
0.0

Ramos (B cell)
0.0
0.0
Lung fibroblast IL-13
0.0
0.0

ionomycin

B lymphocytes PWM
0.0
0.0
Lung fibroblast IFN
0.0
0.0

gamma

B lymphocytes
0.0
0.0
Dermal fibroblast
0.0
0.0

CD40L and IL-4

CCD1070 rest

EOL-1 dbcAMP
0.0
0.0
Dermal fibroblast
0.0
0.0

CCD1070 TNF alpha

EOL-1 dbcAMP
0.0
0.0
Dermal fibroblast
0.0
0.0

PMA/ionomycin

CCD1070 IL-1beta

Dendritic cells none
0.0
0.0
Dermal fibroblast IFN
0.0
0.0

gamma

Dendritic cells LPS
0.0
0.0
Dermal fibroblast IL-4
0.0
0.0

Dendritic cells anti-
0.0
0.0
IBD Colitis 2
20.7
16.4

CD40

Monocytes rest
0.0
0.0
IBD Crohn's
0.0
0.0

Monocytes LPS
0.0
0.0
Colon
3.5
0.0

Macrophages rest
0.0
0.0
Lung
0.0
0.0

Macrophages LPS
0.0
0.0
Thymus
0.0
0.0

HUVEC none
0.0
35.4
Kidney
0.0
0.0

HUVEC starved
0.0
0.0

[0660] CNS_neurodegeneration_v1.0 Summary: Ag2596 Expression of the CG55769-01 gene is low/undetectable (CTs>35) across all of the samples on this panel (data not shown).

[0661] Panel 1.3D Summary: Ag1736/Ag2596 Two experiments with two different probe and primer sets show highest expression of the CG55769-01 gene in the spleen and a colon cancer cell line. Both runs show low but significant expression in a liver adenocarcinoma cell line. Thus, expression of this gene could be used to differentiate between these samples and other samples on this panel.

[0662] Panel 2.2 Summary: Ag1736/Ag2596 Results from two experiments using different probe/primer sets are in reasonable agreement. Significant expression of the CG55769-01 gene is seen exclusively in a metastatic breast cancer sample. Therefore, expression of this gene may be used to distinguish metastatic breast cancers from the other samples on this panel. Furthermore, therapeutic modulation of the activity of the GPCR encoded by this gene may be beneficial in the treatment of metastatic breast cancer.

[0663] Panel 4D Summary: Ag1736/Ag2596 Two experiments with two different probe and primer sets show significant expression of the CG55769-01 gene restricted to liver cirrhosis (CTs=31-34). Furthermore, expression of this gene is not detected in normal liver in Panel 1.3D, suggesting that its expression is unique to liver cirrhosis. This gene encodes a putative GPCR; therefore, antibodies or small molecule therapeutics could reduce or inhibit fibrosis that occurs in liver cirrhosis. In addition, antibodies to this putative GPCR could also be used for the diagnosis of liver cirrhosis (Mark et al., G protein modulation of recombinant P/Q-type calcium channels by regulators of G protein signalling proteins. J. Physiol. 528 Pt 1:65-77, 2000).

[0664] AA. CG92727-02/GMAC010760 A: Olfactory Receptor

[0665] Expression of gene CG92727-02 was assessed using the primer-probe sets Ag1806 and Ag1707, described in Tables AAA and AAB. Results of the RTQ-PCR runs are shown in Tables AAC, AAD, and AAE.

122TABLE AAAProbe Name Ag1806StartSEQ IDPrimersSequencesLengthPositionNO:Forward5′-tcttggtctttgtgctgatcct-3′2276289ProbeTET-5′-tcatcctccctggaaattttctcatt-3′-TAMRA26112290Reverse5′-agggtctgaccttatggtgaaa-3′22140291

[0666]

123

TABLE AAB

Probe Name Ag1707

Start
SEQ ID

Primers
Sequences
Length
Position
NO:

Forward
5′-agcttctgaagggaagaacaag-3′
22
686
292

Probe
TET-5′-ccacgtgcaccactcgtgtcattatt-3′-TAMRA
26
715
293

Reverse
5′-atgaagatagcaggtccaaaca-3′
22
751
294

[0667]

124

TABLE AAC

General_screening_panel_v1.5

Rel. Exp. (%) Ag1806, Run

Rel. Exp. (%) Ag1806, Run

Tissue Name
228714747
Tissue Name
228714747

Adipose
0.0
Renal ca. TK-10
0.0

Melanoma* Hs688(A).T
0.0
Bladder
0.0

Melanoma* Hs688(B).T
0.0
Gastric ca. (liver met.)
0.0

NCI-N87

Melanoma* M14
0.0
Gastric ca. KATO III
0.0

Melanoma* LOXIMVI
0.0
Colon ca. SW-948
0.0

Melanoma* SK-MEL-5
0.0
Colon ca. SW480
0.4

Squamous cell
0.0
Colon ca.* (SW480 met)
0.0

carcinoma SCC-4

SW620

Testis Pool
100.0
Colon ca. HT29
0.0

Prostate ca.* (bone met)
0.4
Colon ca. HCT-116
0.0

PC-3

Prostate Pool
0.4
Colon ca. CaCo-2
0.0

Placenta
0.0
Colon cancer tissue
0.4

Uterus Pool
0.0
Colon ca. SW1116
0.0

Ovarian ca. OVCAR-3
0.0
Colon ca. Colo-205
0.0

Ovarian ca. SK-OV-3
0.0
Colon ca. SW-48
0.0

Ovarian ca. OVCAR-4
0.9
Colon Pool
0.0

Ovarian ca. OVCAR-5
0.0
Small Intestine Pool
0.0

Ovarian ca. IGROV-1
0.0
Stomach Pool
0.0

Ovarian ca. OVCAR-8
0.0
Bone Marrow Pool
0.0

Ovary
0.0
Fetal Heart
0.0

Breast ca. MCF-7
0.4
Heart Pool
0.4

Breast ca. MDA-MB-
0.0
Lymph Node Pool
0.0

231

Breast ca. BT 549
0.0
Fetal Skeletal Muscle
0.0

Breast ca. T47D
0.0
Skeletal Muscle Pool
0.4

Breast ca. MDA-N
0.0
Spleen Pool
0.0

Breast Pool
0.0
Thymus Pool
0.7

Trachea
0.0
CNS cancer (glio/astro)
0.0

U87-MG

Lung
0.0
CNS cancer (glio/astro) U-
0.0

118-MG

Fetal Lung
0.0
CNS cancer (neuro;met)
0.0

SK-N-AS

Lung ca. NCI-N417
0.0
CNS cancer (astro) SF-539
0.0

Lung ca. LX-1
0.0
CNS cancer (astro) SNB-75
1.4

Lung ca. NCI-H146
0.0
CNS cancer (glio) SNB-19
0.0

Lung ca. SHP-77
0.0
CNS cancer (glio) SF-295
0.0

Lung ca. A549
0.0
Brain (Amygdala) Pool
0.7

Lung ca. NCI-H526
0.0
Brain (cerebellum)
0.0

Lung ca. NCI-H23
0.0
Brain (fetal)
0.0

Lung ca. NCI-H460
26.4
Brain (Hippocampus) Pool
1.5

Lung ca. HOP-62
0.0
Cerebral Cortex Pool
1.0

Lung ca. NCI-H522
0.0
Brain (Substantia nigra)
1.9

Pool

Liver
0.0
Brain (Thalamus) Pool
0.9

Fetal Liver
0.0
Brain (whole)
0.0

Liver ca. HepG2
0.0
Spinal Cord Pool
0.7

Kidney Pool
0.0
Adrenal Gland
0.0

Fetal Kidney
0.0
Pituitary gland Pool
0.0

Renal ca. 786-0
0.0
Salivary Gland
0.0

Renal ca. A498
0.0
Thyroid (female)
0.0

Renal ca. ACHN
0.0
Pancreatic ca. CAPAN2
0.0

Renal ca. UO-31
0.0
Pancreas Pool
0.0

[0668]

125

TABLE AAD

Panel 1.3D

Rel. Exp. (%) Ag1806, Run

Rel. Exp. (%) Ag1806, Run

Tissue Name
165975009
Tissue Name
165975009

Liver adenocarcinoma
0.0
Kidney (fetal)
0.0

Pancreas
0.0
Renal ca. 786-0
0.0

Pancreatic ca. CAPAN 2
0.0
Renal ca. A498
0.0

Adrenal gland
0.0
Renal ca. RXF 393
0.0

Thyroid
0.0
Renal ca. ACHN
0.0

Salivary gland
0.0
Renal ca. UO-31
0.0

Pituitary gland
0.0
Renal ca. TK-10
0.0

Brain (fetal)
0.0
Liver
0.0

Brain (whole)
3.7
Liver (fetal)
0.0

Brain (amygdala)
0.0
Liver ca. (hepatoblast)
0.0

HepG2

Brain (cerebellum)
0.0
Lung
0.0

Brain (hippocampus)
0.7
Lung (fetal)
0.0

Brain (substantia nigra)
0.0
Lung ca. (small cell) LX-1
0.0

Brain (thalamus)
0.0
Lung ca. (small cell)
0.0

NCI-H69

Cerebral Cortex
0.0
Lung ca. (s.cell var.)
0.0

SHP-77

Spinal cord
2.9
Lung ca. (large cell)NCI-
0.0

H460

glio/astro U87-MG
0.0
Lung ca. (non-sm. cell)
0.0

A549

glio/astro U-118-MG
0.0
Lung ca. (non-s.cell)
0.0

NCI-H23

astrocytoma SW1783
0.0
Lung ca. (non-s.cell)
0.0

HOP-62

neuro*; met SK-N-AS
0.0
Lung ca. (non-s.cl) NCI-
0.0

H522

astrocytoma SF-539
0.0
Lung ca. (squam.) SW
0.0

900

astrocytoma SNB-75
0.0
Lung ca. (squam.) NCI-
0.0

H596

glioma SNB-19
0.0
Mammary gland
0.0

glioma U251
0.0
Breast ca.* (pl.ef) MCF-7
0.0

glioma SF-295
0.0
Breast ca.* (pl.ef) MDA-
0.0

MB-231

Heart (fetal)
0.0
Breast ca.* (pl.ef) T47D
0.0

Heart
0.0
Breast ca. BT-549
0.0

Skeletal muscle (fetal)
0.0
Breast ca. MDA-N
0.0

Skeletal muscle
0.0
Ovary
0.0

Bone marrow
0.0
Ovarian ca. OVCAR-3
0.0

Thymus
1.3
Ovarian ca. OVCAR-4
0.0

Spleen
6.4
Ovarian ca. OVCAR-5
0.0

Lymph node
1.1
Ovarian ca. OVCAR-8
0.0

Colorectal
1.1
Ovarian ca. IGROV-1
0.0

Stomach
0.0
Ovarian ca.* (ascites)
0.0

SK-OV-3

Small intestine
0.0
Uterus
0.0

Colon ca. SW480
0.0
Plancenta
0.0

Colon ca.* SW620(SW480
0.0
Prostate
1.3

met)

Colon ca. HT29
0.0
Prostate ca.* (bone
0.0

met)PC-3

Colon ca. HCT-116
0.0
Testis
100.0

Colon ca. CaCo-2
0.0
Melanoma Hs688(A).T
0.0

Colon ca.
0.0
Melanoma* (met)
0.0

tissue(ODO3866)

Hs688(B).T

Colon ca. HCC-2998
0.0
Melanoma UACC-62
0.0

Gastric ca.* (liver met)
0.0
Melanoma M14
0.0

NCI-N87

Bladder
0.0
Melanoma LOX IMVI
0.0

Trachea
0.0
Melanoma* (met) SK-
0.0

MEL-5

Kidney
0.0
Adipose
0.0

[0669]

126

TABLE AAE

Panel 4D

Rel. Exp. (%)
Rel. Exp. (%)

Rel. Exp. (%)
Rel. Exp. (%)

Ag1707, Run
Ag1806, Run

Ag1707, Run
Ag1806, Run

Tissue Name
165768285
165812559
Tissue Name
165768285
165812559

Secondary Th1 act
0.0
0.0
HUVEC IL-1beta
0.0
0.0

Secondary Th2 act
0.0
1.6
HUVEC IFN gamma
0.0
0.0

Secondary Tr1 act
0.0
0.4
HUVEC TNF alpha +
0.0
0.0

IFN gamma

Secondary Th1 rest
0.0
0.0
HUVEC TNF alpha +
0.0
0.0

IL4

Secondary Th2 rest
0.0
0.0
HUVEC IL-11
0.0
0.0

Secondary Tr1 rest
0.0
0.0
Lung Microvascular
0.0
0.0

EC none

Primary Th1 act
0.0
0.0
Lung Microvascular
0.0
0.0

EC TNF alpha + IL-

1beta

Primary Th2 act
0.0
2.9
Microvascular
0.0
0.0

Dermal EC none

Primary Tr1 act
2.5
0.0
Microsvasular Dermal
0.0
0.0

EC TNF alpha + IL-

1beta

Primary Th1 rest
0.0
3.2
Bronchial epithelium
0.0
0.0

TNF alpha + IL1beta

Primary Th2 rest
0.0
0.0
Small airway
0.0
0.0

epithelium none

Primary Tr1 rest
1.2
0.0
Small airway
0.0
0.0

epithelium TNF alpha +

IL-1beta

CD45RA CD4
0.0
0.0
Coronery artery SMC
0.0
0.0

lymphocyte act

rest

CD45RO CD4
0.0
0.0
Coronery artery SMC
0.0
0.0

lymphocyte act

TNF alpha + IL-1beta

CD8 lymphocyte act
0.0
0.0
Astrocytes rest
0.0
0.0

Secondary CD8
0.0
0.0
Astrocytes TNF alpha +
0.0
0.0

lymphocyte rest

IL-1beta

Secondary CD8
0.0
0.0
KU-812 (Basophil)
0.0
0.0

lymphocyte act

rest

CD4 lymphocyte
0.0
0.0
KU-812 (Basophil)
0.0
0.0

none

PMA/ionomycin

2ry
0.0
0.0
CCD1106
0.0
0.0

Th1/Th2/Tr1_anti-

(Keratinocytes) none

CD95 CH11

LAK cells rest
0.0
0.0
CCD1106
0.0
0.0

(Keratinocytes)

TNF alpha + IL-1beta

LAK cells IL-2
1.7
0.0
Liver cirrhosis
71.7
87.7

LAK cells IL-2 + IL-
0.0
0.0
Lupus kidney
0.0
0.0

12

LAK cells IL-2 + IFN
0.0
0.0
NCI-H292 none
52.5
68.3

gamma

LAK cells IL-2 + IL-
0.0
0.0
NCI-H292 IL-4
81.8
100.0

18

LAK cells
0.0
0.0
NCI-H292 IL-9
100.0
77.9

PMA/ionomycin

NK Cells IL-2 rest
0.0
0.0
NCI-H292 IL-13
33.2
28.1

Two Way MLR 3
0.0
0.0
NCI-H292 IFN
16.3
15.3

day

gamma

Two Way MLR 5
0.0
0.0
HPAEC none
0.0
0.0

day

Two Way MLR 7
0.0
0.0
HPAEC TNF alpha +
0.0
0.0

day

IL-1beta

PBMC rest
0.0
0.0
Lung fibroblast none
0.0
0.0

PBMC PWM
0.0
0.0
Lung fibroblast TNF
0.0
0.0

alpha + IL-1beta

PBMC PHA-L
0.0
0.0
Lung fibroblast IL-4
0.0
0.0

Ramos (B cell) none
0.0
0.0
Lung fibroblast IL-9
0.0
0.0

Ramos (B cell)
0.0
0.0
Lung fibroblast IL-13
0.0
0.0

ionomycin

B lymphocytes PWM
1.4
0.0
Lung fibroblast IFN
0.0
0.0

gamma

B lymphocytes
0.0
1.4
Dermal fibroblast
0.0
0.0

CD40L and IL-4

CCD1070 rest

EOL-1 dbcAMP
0.0
0.0
Dermal fibroblast
0.0
13.6

CCD1070 TNF alpha

EOL-1 dbcAMP
0.0
0.0
Dermal fibroblast
1.2
0.0

PMA/ionomycin

CCD1070 IL-1beta

Dendritic cells none
0.0
0.0
Dermal fibroblast IFN
0.0
0.0

gamma

Dendritic cells LPS
0.0
0.0
Dermal fibroblast IL-4
0.0
0.0

Dendritic cells anti-
0.0
0.0
IBD Colitis 2
20.2
42.3

CD40

Monocytes rest
0.0
0.0
IBD Crohn's
4.5
15.7

Monocytes LPS
1.7
0.0
Colon
12.6
2.8

Macrophages rest
0.0
0.0
Lung
4.9
4.6

Macrophages LPS
0.0
0.0
Thymus
1.5
0.0

HUVEC none
0.0
0.0
Kidney
4.1
6.8

HUVEC starved
0.0
0.0

[0670] CNS_neurodegeneration_v1.0 Summary: Ag1806 Expression of the CG92727-01 gene is low/undetected (CT>35) in all samples in this panel. (Data not shown.) General_screening_panel_v1.5 Summary: Ag1806 Expression of the CG92727-01 gene is restricted to the testis and a lung cancer cell line in this panel (CTs=31-34). This expression profile suggests that the protein encoded by the CG92727-01 gene may be involved in fertility. Therefore, therapeutic modulation of the function or expression of this gene product may be effective in treating fertility related disorders. Furthermore, the presence of the transcript in a lung cancer cell line indicates that the expression of this gene could be used to differentiate lung cancer cell lines from other samples in this panel. Therapeutic modulation of the function or expression of the CG92727-01 protein product may also be effective in the treatment of lung cancer.

[0671] Panel 1.3D Summary: Ag1806 Expression of the CG92727-01 gene in this panel is restricted to the testis (CT=31). This is the same expression profile seen in General_screening_panel_v1.5. Please see that panel for discussion of potential utility of this gene.

[0672] Panel 2.2 Summary: Ag1806 Expression of the CG92727-01 gene is low/undetected (CT>35) in all samples in this panel. (Data not shown.)

[0673] Panel 4D Summary: Ag1806 The CG92727-01 gene is constitutively expressed in the NCI-H292 mucoepidermoid cell line (CTs=32-33). In comparison, expression in the normal lung is low. The expression of the transcript in the NCI-H292 cell line, often used as a model for airway epithelium, suggests that this transcript may be important in the proliferation or activation of airway epithelium. Therefore, therapeutics designed with the GPCR encoded by the transcript could be important in the treatment of diseases that exhibit lung airway inflammation such as asthma and COPD.

[0674] This transcript is also expressed in liver cirrhosis and colitis. Normal liver and colon do not express this transcript (see panel 1.3 and 2.2 for liver) suggesting that expression may be specific to cirrhosis. The transcript or the protein encoded for by the transcript could be used diagnostically to identify liver cirrhosis or colitis. Furthermore, the protein encoded by this transcript could be used to design therapeutics against liver cirrhosis or colitis.

[0675] AB. CG55993-01: Olfactory Receptor

[0676] Expression of gene CG55993-01 was assessed using the primer-probe sets Ag2322, Ag2365 and Ag2350, described in Tables ABA, ABB and ABC. Results of the RTQ-PCR runs are shown in Tables ABD, ABE, ABF, and ABG.

127TABLE ABAProbe Name Ag2322StartSEQ IDPrimersSequencesLengthPositionNO:Forward5′-atcccactgtgcttcatgtatc-3′22162295ProbeTET-5′-atcccgggcaactgcacaattctttt-3′-TAMRA26192296Reverse5′-agtgagcgctctgttttaatga-3′22220297

[0677]

128

TABLE ABB

Probe Name Ag2365

Start
SEQ ID

Primers
Sequences
Length
Position
NO:

Forward
5′-atcccactgtgcttcatgtatc-3′
22
162
298

Probe
TET-5′-atcccgggcaactgcacaattctttt-3′-TAMRA
26
192
299

Reverse
5′-agtgagcgctctgttttaatga-3′
22
220
300

[0678]

129

TABLE ABC

Probe Name Ag2350

Start
SEQ ID

Primers
Sequences
Length
Position
NO:

Forward
5′-atcccactgtgcttcatgtatc-3′
22
162
301

Probe
TET-5′-atcccgggcaactgcacaattctttt-3′-TAMRA
26
192
302

Reverse
5′-agtgagcgctctgttttaatga-3′
22
220
303

[0679]

130

TABLE ABD

AI_comprehensive panel_v1.0

Rel. Exp. (%) Ag2322,

Rel. Exp. (%) Ag2322,

Tissue Name
Run 229743870
Tissue Name
Run 229743870

110967 COPD-F
2.2
112427 Match Control
0.0

Psoriasis-F

110980 COPD-F
0.0
112418 Psoriasis-M
0.0

110968 COPD-M
0.0
112723 Match Control
1.5

Psoriasis-M

110977 COPD-M
0.0
112419 Psoriasis-M
0.0

110989 Emphysema-F
0.0
112424 Match Control
3.0

Psoriasis-M

110992 Emphysema-F
0.0
112420 Psoriasis-M
0.0

110993 Emphysema-F
0.0
112425 Match Control
2.3

Psoriasis-M

110994 Emphysema-F
0.0
104689 (MF) OA Bone-
0.0

Backus

110995 Emphysema-F
0.0
104690 (MF) Adj “Normal”
0.0

Bone-Backus

110996 Emphysema-F
1.7
104691 (MF) OA
0.0

Synovium-Backus

110997 Asthma-M
2.5
104692 (BA) OA Cartilage-
0.0

Backus

111001 Asthma-F
0.0
104694 (BA) OA Bone-
0.0

Backus

111002 Asthma-F
0.0
104695 (BA) Adj “Normal”
0.0

Bone-Backus

111003 Atopic Asthma-F
0.0
104696 (BA) OA Synovium-
0.0

Backus

111004 Atopic Asthma-F
0.0
104700 (SS) OA Bone-
0.0

Backus

111005 Atopic Asthma-F
0.0
104701 (SS) Adj “Normal”
0.0

Bone-Backus

111006 Atopic Asthma-F
0.0
104702 (SS) OA Synovium-
0.0

Backus

111417 Allergy-M
0.0
117093 OA Cartilage Rep7
0.0

112347 Allergy-M
4.6
112672 OA Bone5
0.0

112349 Normal Lung-F
4.5
112673 OA Synovium5
0.0

112357 Normal Lung-F
2.0
112674 OA Synovial Fluid
0.0

cells5

112354 Normal Lung-M
0.0
117100 OA Cartilage Rep14
0.0

112374 Crohns-F
0.0
112756 OA Bone9
100.0

112389 Match Control
0.0
112757 OA Synovium9
0.0

Crohns-F

112375 Crohns-F
0.0
112758 OA Synovial Fluid
0.0

Cells9

112732 Match Control
0.0
117125 RA Cartilage Rep2
0.0

Crohns-F

112725 Crohns-M
0.0
113492 Bone2 RA
1.7

112387 Match Control
2.3
113493 Synovium2 RA
0.0

Crohns-M

112378 Crohns-M
11.3
113494 Syn Fluid Cells RA
0.0

112390 Match Control
0.0
113499 Cartilage4 RA
0.0

Crohns-M

112726 Crohns-M
2.6
113500 Bone4 RA
0.0

112731 Match Control
0.0
113501 Synovium4 RA
0.0

Crohns-M

112380 Ulcer Col-F
0.0
113502 Syn Fluid Cells4 RA
0.0

112734 Match Control
0.0
113495 Cartilage3 RA
0.0

Ulcer Col-F

112384 Ulcer Col-F
1.9
113496 Bone3 RA
0.0

112737 Match Control
0.0
113497 Synovium3 RA
0.0

Ulcer Col-F

112386 Ulcer Col-F
0.0
113498 Syn Fluid Cells3 RA
0.0

112738 Match Control
0.0
117106 Normal Cartilage
0.0

Ulcer Col-F

Rep20

112381 Ulcer Col-M
0.0
113663 Bone3 Normal
2.6

112735 Match Control
0.0
113664 Synovium3 Normal
2.6

Ulcer Col-M

112382 Ulcer Col-M
0.0
113665 Syn Fluid Cells3
0.0

Normal

112394 Match Control
2.1
117107 Normal Cartilage
0.0

Ulcer Col-M

Rep22

112383 Ulcer Col-M
1.4
113667 Bone4 Normal
0.0

112736 Match Control
0.0
113668 Synovium4 Normal
2.2

Ulcer Col-M

112423 Psoriasis-F
0.0
113669 Syn Fluid Cells4
2.0

Normal

[0680]

131

TABLE ABE

Panel 1.3D

Rel. Exp. (%)
Rel. Exp. (%)

Rel. Exp. (%)
Rel. Exp. (%)

Ag2322, Run
Ag2350, Run

Ag2322, Run
Ag2350, Run

Tissue Name
165627868
165974845
Tissue Name
165627868
165974845

Liver
0.0
0.0
Kidney (fetal)
0.0
0.0

adenocarcinoma

Pancreas
0.0
0.0
Renal ca. 786-0
0.0
0.0

Pancreatic ca.
0.0
0.0
Renal ca. A498
0.0
0.0

CAPAN 2

Adrenal gland
0.0
0.0
Renal ca. RXF
0.0
0.0

393

Thyroid
0.0
0.0
Renal ca. ACHN
0.0
0.0

Salivary gland
0.0
0.0
Renal ca. UO-31
0.0
0.0

Pituitary gland
0.0
7.6
Renal ca. TK-10
0.0
0.0

Brain (fetal)
0.0
0.0
Liver
0.0
0.0

Brain (whole)
0.0
0.0
Liver (fetal)
0.0
0.0

Brain (amygdala)
0.0
0.0
Liver ca.
0.0
0.0

(hepatoblast)

HepG2

Brain (cerebellum)
0.0
0.0
Lung
0.0
0.0

Brain (hippocampus)
0.0
0.0
Lung (fetal)
0.0
0.0

Brain (substantia
0.0
0.0
Lung ca. (small
0.0
0.0

nigra)

cell) LX-1

Brain (thalamus)
0.0
0.0
Lung ca. (small
0.0
31.2

cell) NCI-H69

Cerebral Cortex
0.0
0.0
Lung ca. (s. cell
100.0
100.0

var.) SHP-77

Spinal cord
3.4
0.0
Lung ca. (large
0.0
0.0

cell) NCI-H460

glio/astro U87-MG
0.0
0.0
Lung ca. (non-
0.0
0.0

sm. cell) A549

glio/astro U-118-
0.0
0.0
Lung ca. (non-
0.0
0.0

MG

s. cell) NCI-H23

astrocytoma
0.0
0.0
Lung ca. (non-
0.0
0.0

SW1783

s. cell) HOP-62

neuro*; met SK-N-
0.0
0.0
Lung ca. (non-
0.0
0.0

AS

s. cl) NCI-H522

astrocytoma SF-539
0.0
0.0
Lung ca.
0.0
0.0

(squam.) SW 900

astrocytoma SNB-75
0.0
0.0
Lung ca.
16.6
15.5

(squam.) NCI-

H596

glioma SNB-19
2.5
0.0
Mammary gland
0.0
0.0

glioma U251
0.0
0.0
Breast ca.*
0.0
0.0

(pl. ef) MCF-7

glioma SF-295
0.0
0.0
Breast ca.*
0.0
0.0

(pl. ef) MDA-

MB-231

Heart (fetal)
0.0
0.0
Breast ca.*
0.0
0.0

(pl. ef) T47D

Heart
0.0
0.0
Breast ca. BT-
0.0
0.0

549

Skeletal muscle
0.0
0.0
Breast ca. MDA-N
0.0
0.0

(fetal)

Skeletal muscle
0.0
0.0
Ovary
0.0
0.0

Bone marrow
0.0
0.0
Ovarian ca.
1.7
0.0

OVCAR-3

Thymus
0.0
0.0
Ovarian ca.
0.0
0.0

OVCAR-4

Spleen
0.0
0.0
Ovarian ca.
0.0
0.0

OVCAR-5

Lymph node
0.0
0.0
Ovarian ca.
0.0
4.8

OVCAR-8

Colorectal
1.4
0.0
Ovarian ca.
0.0
0.0

IGROV-1

Stomach
0.0
0.0
Ovarian ca.*
0.0
0.0

(ascites) SK-OV-3

Small intestine
0.0
0.0
Uterus
0.0
0.0

Colon ca. SW480
0.0
0.0
Plancenta
0.0
0.0

Colon ca.*
0.0
0.0
Prostate
0.0
0.0

SW620(SW480 met)

Colon ca. HT29
0.0
0.0
Prostate ca.*
0.0
0.0

(bone met)PC-3

Colon ca. HCT-116
0.0
0.0
Testis
0.0
0.0

Colon ca. CaCo-2
0.0
0.0
Melanoma
0.0
0.0

Hs688(A).T

Colon ca.
0.0
0.0
Melanoma*
0.0
0.0

tissue(ODO3866)

(met)

Hs688(B).T

Colon ca. HCC-2998
0.0
0.0
Melanoma
0.0
0.0

UACC-62

Gastric ca.* (liver
3.3
0.0
Melanoma M14
0.0
0.0

met) NCI-N87

Bladder
0.0
6.9
Melanoma LOX
0.0
0.0

IMVI

Trachea
0.0
0.0
Melanoma*
0.0
0.0

(met) SK-MEL-5

Kidney
0.0
0.0
Adipose
0.0
0.0

[0681]

132

TABLE ABF

Panel 2D

Rel. Exp. (%) Ag2350,

Rel. Exp. (%) Ag2350,

Tissue Name
Run 164079723
Tissue Name
Run 164079723

Normal Colon
7.6
Kidney Margin 8120608
0.0

CC Well to Mod Diff
4.1
Kidney Cancer 8120613
0.0

(ODO3866)

CC Margin (ODO3866)
1.5
Kidney Margin 8120614
0.0

CC Gr.2 rectosigmoid
3.0
Kidney Cancer 9010320
0.0

(ODO3868)

CC Margin (ODO3868)
0.0
Kidney Margin 9010321
0.0

CC Mod Diff (ODO3920)
0.0
Normal Uterus
0.0

CC Margin (ODO3920)
7.1
Uterus Cancer 064011
3.7

CC Gr.2 ascend colon
0.0
Normal Thyroid
0.0

(ODO3921)

CC Margin (ODO3921)
0.0
Thyroid Cancer 064010
0.0

CC from Partial Hepatectomy
0.0
Thyroid Cancer A302152
0.0

(ODO4309) Mets

Liver Margin (ODO4309)
0.0
Thyroid Margin A302153
0.0

Colon mets to lung (OD04451-
0.0
Normal Breast
0.0

01)

Lung Margin (OD04451-02)
0.0
Breast Cancer (OD04566)
7.6

Normal Prostate 6546-1
22.4
Breast Cancer (OD04590-
0.0

01)

Prostate Cancer (OD04410)
74.7
Breast Cancer Mets
0.0

(OD04590-03)

Prostate Margin (OD04410)
100.0
Breast Cancer Metastasis
0.0

(OD04655-05)

Prostate Cancer (OD04720-01)
30.1
Breast Cancer 064006
0.0

Prostate Margin (OD04720-02)
38.7
Breast Cancer 1024
0.0

Normal Lung 061010
0.0
Breast Cancer 9100266
0.0

Lung Met to Muscle
0.0
Breast Margin 9100265
1.7

(ODO4286)

Muscle Margin (ODO4286)
0.0
Breast Cancer A209073
7.2

Lung Malignant Cancer
25.5
Breast Margin A2090734
0.0

(OD03126)

Lung Margin (OD03126)
0.0
Normal Liver
0.0

Lung Cancer (OD04404)
6.5
Liver Cancer 064003
0.0

Lung Margin (OD04404)
0.0
Liver Cancer 1025
0.0

Lung Cancer (OD04565)
0.0
Liver Cancer 1026
0.0

Lung Margin (OD04565)
0.0
Liver Cancer 6004-T
0.0

Lung Cancer (OD04237-01)
0.0
Liver Tissue 6004-N
0.0

Lung Margin (OD04237-02)
0.0
Liver Cancer 6005-T
0.0

Ocular Mel Met to Liver
0.0
Liver Tissue 6005-N
0.0

(ODO4310)

Liver Margin (ODO4310)
0.0
Normal Bladder
0.0

Melanoma Mets to Lung
0.0
Bladder Cancer 1023
0.0

(OD04321)

Lung Margin (OD04321)
0.0
Bladder Cancer A302173
6.1

Normal Kidney
0.0
Bladder Cancer
0.0

(OD04718-01)

Kidney Ca, Nuclear grade 2
0.0
Bladder Normal Adjacent
0.0

(OD04338)

(OD04718-03)

Kidney Margin (OD04338)
3.6
Normal Ovary
0.0

Kidney Ca Nuclear grade 1/2
0.0
Ovarian Cancer 064008
0.0

(OD04339)

Kidney Margin (OD04339)
0.0
Ovarian Cancer
0.0

(OD04768-07)

Kidney Ca, Clear cell type
0.0
Ovary Margin (OD04768-
0.0

(OD04340)

08)

Kidney Margin (OD04340)
0.0
Normal Stomach
0.0

Kidney Ca, Nuclear grade 3
1.7
Gastric Cancer 9060358
0.0

(OD04348)

Kidney Margin (OD04348)
0.0
Stomach Margin 9060359
0.0

Kidney Cancer (OD04622-01)
0.0
Gastric Cancer 9060395
0.0

Kidney Margin (OD04622-03)
0.0
Stomach Margin 9060394
0.0

Kidney Cancer (OD04450-01)
0.0
Gastric Cancer 9060397
0.0

Kidney Margin (OD04450-03)
0.0
Stomach Margin 9060396
0.0

Kidney Cancer 8120607
0.0
Gastric Cancer 064005
3.7

[0682]

133

TABLE ABG

Panel 4D

Rel. Exp. (%)
Rel. Exp. (%)

Rel. Exp. (%)
Rel. Exp. (%)

Ag2322, Run
Ag2350, Run

Ag2322, Run
Ag2350, Run

Tissue Name
162360932
164145590
Tissue Name
162360932
164145590

Secondary Th1 act
0.0
0.0
HUVEC IL-1beta
0.0
0.0

Secondary Th2 act
0.0
0.0
HUVEC IFN gamma
0.0
0.0

Secondary Tr1 act
0.0
29.3
HUVEC TNF alpha +
0.0
0.0

IFN gamma

Secondary Th1 rest
0.0
0.0
HUVEC TNF alpha +
0.0
0.0

IL4

Secondary Th2 rest
0.0
0.0
HUVEC IL-11
0.0
0.0

Secondary Tr1 rest
0.0
0.0
Lung Microvascular
0.0
0.0

EC none

Primary Th1 act
0.0
0.0
Lung Microvascular
0.0
0.0

EC TNF alpha + IL-

1beta

Primary Th2 act
0.0
0.0
Microvascular
0.0
0.0

Dermal EC none

Primary Tr1 act
0.0
0.0
Microsvasular Dermal
0.0
0.0

EC TNF alpha + IL-

1beta

Primary Th1 rest
0.0
0.0
Bronchial epithelium
0.0
0.0

TNF alpha + IL1beta

Primary Th2 rest
0.0
0.0
Small airway
0.0
0.0

epithelium none

Primary Tr1 rest
0.0
0.0
Small airway
0.0
0.0

epithelium TNF alpha +

IL-1beta

CD45RA CD4
0.0
0.0
Coronery artery SMC
0.0
0.0

lymphocyte act

rest

CD45RO CD4
0.0
0.0
Coronery artery SMC
0.0
48.0

lymphocyte act

TNF alpha + IL-1beta

CD8 lymphocyte act
0.0
23.3
Astrocytes rest
0.0
0.0

Secondary CD8
0.0
0.0
Astrocytes TNF alpha +
0.0
0.0

lymphocyte rest

IL-1beta

Secondary CD8
21.2
0.0
KU-812 (Basophil)
0.0
0.0

lymphocyte act

rest

CD4 lymphocyte
0.0
0.0
KU-812 (Basophil)
0.0
0.0

none

PMA/ionomycin

2ry
0.0
0.0
CCD1106
0.0
0.0

Th1/Th2/Tr1_anti-

(Keratinocytes) none

CD95 CH11

LAK cells rest
0.0
0.0
CCD1106
0.0
0.0

(Keratinocytes)

TNF alpha + IL-1beta

LAK cells IL-2
0.0
0.0
Liver cirrhosis
100.0
100.0

LAK cells IL-2 + IL-
0.0
0.0
Lupus kidney
0.0
0.0

12

LAK cells IL-2 + IFN
0.0
0.0
NCI-H292 none
0.0
0.0

gamma

LAK cells IL-2 + IL-
0.0
0.0
NCI-H292 IL-4
0.0
0.0

18

LAK cells
0.0
0.0
NCI-H292 IL-9
0.0
00

PMA/ionomycin

NK Cells IL-2 rest
0.0
0.0
NCI-H292 IL-13
0.0
0.0

Two Way MLR 3
0.0
0.0
NCI-H292 IFN
0.0
0.0

day

gamma

Two Way MLR 5
0.0
0.0
HPAEC none
0.0
0.0

day

Two Way MLR 7
0.0
0.0
HPAEC TNF alpha +
0.0
0.0

day

IL-1beta

PBMC rest
0.0
0.0
Lung fibroblast none
0.0
21.0

PBMC PWM
0.0
0.0
Lung fibroblast TNF
0.0
0.0

alpha + IL-1beta

PBMC PHA-L
0.0
0.0
Lung fibroblast IL-4
0.0
0.0

Ramos (B cell) none
0.0
0.0
Lung fibroblast IL-9
0.0
0.0

Ramos (B cell)
0.0
0.0
Lung fibroblast IL-13
0.0
0.0

ionomycin

B lymphocytes PWM
7.1
0.0
Lung fibroblast IFN
0.0
40.1

gamma

B lymphocytes
0.0
0.0
Dermal fibroblast
0.0
0.0

CD40L and IL-4

CCD1070 rest

EOL-1 dbcAMP
0.0
0.0
Dermal fibroblast
0.0
0.0

CCD1070 TNF alpha

EOL-1 dbcAMP
0.0
0.0
Dermal fibroblast
0.0
0.0

PMA/ionomycin

CCD1070 IL-1beta

Dendritic cells none
0.0
0.0
Dermal fibroblast IFN
0.0
0.0

gamma

Dendritic cells LPS
0.0
0.0
Dermal fibroblast IL-4
0.0
0.0

Dendritic cells anti-
0.0
0.0
IBD Colitis 2
43.2
19.2

CD40

Monocytes rest
0.0
0.0
IBD Crohn's
0.0
15.3

Monocytes LPS
0.0
0.0
Colon
0.0
0.0

Macrophages rest
0.0
0.0
Lung
0.0
0.0

Macrophages LPS
0.0
0.0
Thymus
0.0
19.1

HUVEC none
0.0
0.0
Kidney
0.0
0.0

HUVEC starved
0.0
0.0

[0683] AI_comprehensive panel_v1.0 Summary: Ag2322 Low but significant expression of the CG55993-03 gene is detected in an osteoarthritic bone sample (CT=33). The protein encoded by this gene is homologous to a G-protein coupled receptors. These receptors have been shown to be involved in calcium homeostasis and control (reference 1). Changes in bone tissue (eg subchondral bone hardening) are identified in osteoarthritis. Therefore, therapeutic modulation of this gene product may ameliorate non-homeostatic bone-related changes found in osteoarthritis (Gardella and Juppner, Molecular properties of the PTH/PTHrP receptor. Trends Endocrinol Metab 12(5):210-7, 2001).

[0684] CNS_neurodegeneration_v1.0 Summary: Ag2365 Data from one experiment with this probe and primer set and the CG55993-03 gene is not included due to a probable probe failure.

[0685] Panel 1.3D Summary: Ag2322/2350 In two runs with the same probe and primer set, highest expression of the CG55993-03 gene is seen in a sample derived from a small cell lung cancer cell line (SHP-77) (CTs=32-33). There is apparent expression in other small cell lung cancer cell lines as well. Thus, the expression of this gene could be used to distinguish SHP-77 cells and other small cell lung cancer cell lines from other samples on the panel. Moreover, therapeutic modulation of this gene, through the use of small molecule drugs, antibodies or protein therapeutics might be of use in the treatment of small cell lung cancer. Please note that a third experiment with the probe and primer Ag2365 showed low/undetectable expression in all samples on this panel (CTs>35). (Data not shown.)

[0686] Panel 2D Summary: Ag2350 The expression of the CG55993-03 gene is highest in a sample derived from normal prostate tissue adjacent to a prostate cancer (CT=32.3). This pattern holds for another matched pair of prostate cancer and normal tissues. There is low to no expression in other tissues, therefore, this gene appears to show prostate specific expression. Thus, the expression of this gene could be used to distinguish prostate derived tissues from other tissues in the panel. Moreover, therapeutic modulation of this gene, through the use of small molecule drugs, antibodies or protein therapeutics might be of benefit in the treatment of prostate cancer.

[0687] Panel 3D Summary: Ag2365 Data from one experiment with this probe and primer set and the CG55993-03 gene is not included due to a probable probe failure.

[0688] Panel 4D Summary: Ag2322/Ag2350 The CG55993-03 transcript is only expressed at significant levels in liver cirrhosis. The transcript is not expressed in normal liver in panel 2 or 1. The transcript or the protein it encodes could be used for detection of liver cirrhosis. The putative GPCR encoded for by this transcript may also play an important role in liver cirrhosis. Therapeutics designed with the protein encoded for by this transcript could be important for maintaining or restoring normal function to the liver undergoing cirrhosis. Please note that a third experiment with the probe and primer Ag2365 showed low/undetectable expression in all samples on this panel (CTs>35). (Data not shown.)

[0689] AC. CG56038-01: Olfactory Receptor

[0690] Expression of gene CG56038-01 was assessed using the primer-probe sets Ag1730 and Ag1830, described in Tables ACA and ACB. Results of the RTQ-PCR runs are shown in Tables ACC and ACD.

134TABLE ACAProbe Name Ag1730StartSEQ IDPrimersSequencesLengthPositionNO:Forward5′-tgatcctgatggactcttgtct-3′22146304ProbeTET-5′-ttcctcagtaacctgtctctggtgga-3′-TAMRA26187305Reverse5′-agtgacagctgaggagtatcca-3′22216306

[0691]

135

TABLE ACB

Probe Name Ag1830

Start
SEQ ID

Primers
Sequences
Length
Position
NO:

Forward
5′-gacaaaatggcatctgtgttct-3′
22
814
307

Probe
TET-5′-ctatgatcatccccatgctgaaccct-3′-TAMRA
26
839
308

Reverse
5′-tgaatgcattctggacttctct-3′
22
886
309

[0692]

136

TABLE ACC

Panel 2.2

Rel. Exp. (%) Ag1730,

Rel. Exp. (%) Ag1730,

Tissue Name
Run 173761867
Tissue Name
Run 173761867

Normal Colon
0.0
Kidney Margin (OD04348)
0.0

Colon cancer (OD06064)
4.8
Kidney malignant cancer
0.0

(OD06204B)

Colon Margin (OD06064)
0.0
Kidney normal adjacent
0.0

tissue (OD06204E)

Colon cancer (OD06159)
0.0
Kidney Cancer (OD04450-
0.0

01)

Colon Margin (OD06159)
0.0
Kidney Margin (OD04450-
0.0

03)

Colon cancer (OD06297-04)
0.0
Kidney Cancer 8120613
0.0

Colon Margin (OD06297-
0.0
Kidney Margin 8120614
0.0

015)

CC Gr.2 ascend colon
0.0
Kidney Cancer 9010320
0.0

(ODO3921)

CC Margin (ODO3921)
0.0
Kidney Margin 9010321
0.0

Colon cancer metastasis
0.0
Kidney Cancer 8120607
0.0

(OD06104)

Lung Margin (OD06104)
0.0
Kidney Margin 8120608
5.3

Colon mets to lung
6.6
Normal Uterus
0.0

(OD04451-01)

Lung Margin (OD04451-02)
0.0
Uterine Cancer 064011
0.0

Normal Prostate
0.0
Normal Thyroid
0.0

Prostate Cancer (OD04410)
0.0
Thyroid Cancer
0.0

Prostate Margin (OD04410)
0.0
Thyroid Cancer A302152
0.0

Normal Ovary
0.0
Thyroid Margin A302153
0.0

Ovarian cancer (OD06283-
0.0
Normal Breast
1.5

03)

Ovarian Margin (OD06283-
0.0
Breast Cancer
6.7

07)

Ovarian Cancer
100.0
Breast Cancer
17.0

Ovarian cancer (OD06145)
0.0
Breast Cancer (OD04590-
0.0

01)

Ovarian Margin (OD06145)
0.0
Breast Cancer Mets
0.0

(OD04590-03)

Ovarian cancer (OD06455-
0.0
Breast Cancer Metastasis
0.0

03)

Ovarian Margin (OD06455-
0.0
Breast Cancer
0.0

07)

Normal Lung
0.0
Breast Cancer 9100266
0.0

Invasive poor diff. lung
0.0
Breast Margin 9100265
0.0

adeno (ODO4945-01

Lung Margin (ODO4945-03)
0.0
Breast Cancer A209073
0.0

Lung Malignant Cancer
0.0
Breast Margin A2090734
0.0

(OD03126)

Lung Margin (OD03126)
0.0
Breast cancer (OD06083)
22.4

Lung Cancer (OD05014A)
0.0
Breast cancer node
7.3

metastasis (OD06083)

Lung Margin (OD05014B)
0.0
Normal Liver
6.7

Lung cancer (OD06081)
0.0
Liver Cancer 1026
0.0

Lung Margin (OD06081)
0.0
Liver Cancer 1025
15.7

Lung Cancer (OD04237-01)
0.0
Liver Cancer 6004-T
0.0

Lung Margin (OD04237-02)
0.0
Liver Tissue 6004-N
0.0

Ocular Mel Met to Liver
0.0
Liver Cancer 6005-T
0.0

(ODO4310)

Liver Margin (ODO4310)
9.3
Liver Tissue 6005-N
0.0

Melanoma Metastasis
0.0
Liver Cancer
0.0

Lung Margin (OD04321)
0.0
Normal Bladder
0.0

Normal Kidney
0.0
Bladder Cancer
0.0

Kidney Ca, Nuclear grade 2
0.0
Bladder Cancer
0.0

(OD04338)

Kidney Margin (OD04338)
0.0
Normal Stomach
0.0

Kidney Ca Nuclear grade 1/2
0.0
Gastric Cancer 9060397
0.0

(OD04339)

Kidney Margin (OD04339)
0.0
Stomach Margin 9060396
0.0

Kidney Ca, Clear cell type
0.0
Gastric Cancer 9060395
28.1

(OD04340)

Kidney Margin (OD04340)
0.0
Stomach Margin 9060394
0.0

Kidney Ca, Nuclear grade 3
0.0
Gastric Cancer 064005
0.0

(OD04348)

[0693]

137

TABLE ACD

Panel 4D

Rel. Exp. (%) Ag1830,

Rel. Exp. (%) Ag1830,

Tissue Name
Run 165810392
Tissue Name
Run 165810392

Secondary Th1 act
3.3
HUVEC IL-1 beta
0.0

Secondary Th2 act
9.5
HUVEC IFN gamma
0.0

Secondary Tr1 act
2.1
HUVEC TNF alpha + IFN
0.0

gamma

Secondary Th1 rest
0.0
HUVEC TNF alpha + IL4
0.0

Secondary Th2 rest
0.0
HUVEC IL-11
0.0

Secondary Tr1 rest
3.9
Lung Microvascular EC none
13.6

Primary Th1 act
0.0
Lung Microvascular EC
9.2

TNF alpha + IL-1 beta

Primary Th2 act
0.0
Microvascular Dermal EC none
44.4

Primary Tr1 act
2.2
Microsvasular Dermal EC
11.2

TNF alpha + IL-1 beta

Primary Th1 rest
9.5
Bronchial epithelium TNF alpha +
4.2

IL1 beta

Primary Th2 rest
2.4
Small airway epithelium none
3.5

Primary Tr1 rest
0.7
Small airway epithelium
0.0

TNF alpha + IL-1 beta

CD45RA CD4 lymphocyte
2.9
Coronery artery SMC rest
0.0

act

CD45RO CD4 lymphocyte
0.0
Coronery artery SMC TNF alpha +
0.0

act

IL-1 beta

CD8 lymphocyte act
4.6
Astrocytes rest
0.0

Secondary CD8
5.3
Astrocytes TNF alpha + IL-1 beta
0.0

lymphocyte rest

Secondary CD8
5.5
KU-812 (Basophil) rest
0.0

lymphocyte act

CD4 lymphocyte none
4.7
KU-812 (Basophil)
0.0

PMA/ionomycin

2ry Th1/Th2/Tr1_anti-
5.2
CCD1106 (Keratinocytes) none
0.0

CD95 CH11

LAK cells rest
3.8
CCD1106 (Keratinocytes)
3.3

TNF alpha + IL-1 beta

LAK cells IL-2
11.3
Liver cirrhosis
100.0

LAK cells IL-2 + IL-12
3.0
Lupus kidney
0.0

LAK cells IL-2 + IFN
8.7
NCI-H292 none
0.0

gamma

LAK cells IL-2 + IL-18
10.7
NCI-H292 IL-4
0.0

LAK cells
9.2
NCI-H292 IL-9
0.0

PMA/ionomycin

NK Cells IL-2 rest
0.0
NCI-H292 IL-13
0.0

Two Way MLR 3 day
18.6
NCI-H292 IFN gamma
0.0

Two Way MLR 5 day
0.0
HPAEC none
0.0

Two Way MLR 7 day
4.4
HPAEC TNF alpha + IL-1 beta
3.5

PBMC rest
3.5
Lung fibroblast none
0.0

PBMC PWM
0.0
Lung fibroblast TNF alpha + IL-
16.6

1 beta

PBMC PHA-L
4.7
Lung fibroblast IL-4
0.0

Ramos (B cell) none
0.0
Lung fibroblast IL-9
3.8

Ramos (B cell) ionomycin
0.0
Lung fibroblast IL-13
0.0

B lymphocytes PWM
2.9
Lung fibroblast IFN gamma
0.0

B lymphocytes CD40L
5.0
Dermal fibroblast CCD1070 rest
0.0

and IL-4

EOL-1 dbcAMP
0.0
Dermal fibroblast CCD1070
7.8

TNF alpha

EOL-1 dbcAMP
21.9
Dermal fibroblast CCD1070 IL-
3.3

PMA/ionomycin

1 beta

Dendritic cells none
3.9
Dermal fibroblast IFN gamma
0.0

Dendritic cells LPS
8.6
Dermal fibroblast IL-4
3.9

Dendritic cells anti-CD40
4.1
IBD Colitis 2
36.6

Monocytes rest
0.0
IBD Crohn's
2.5

Monocytes LPS
9.8
Colon
6.3

Macrophages rest
0.0
Lung
2.5

Macrophages LPS
0.0
Thymus
4.2

HUVEC none
0.0
Kidney
9.9

HUVEC starved
0.0

[0694] Panel 1.3D Summary: Ag1730 Expression of the CG56038-01 gene is low/undetectable (CTs>35) across all of the samples on this panel (data not shown).

[0695] Panel 2.2 Summary: Ag1730 Significant expression of the CG56038-01 gene is seen exclusively in an ovarian cancer sample (CT=33.1). Therefore, expression of this gene may be used to distinguish ovarian cancers from the other samples on this panel. Furthermore, therapeutic modulation of the activity of the GPCR encoded by this gene may be beneficial in the treatment of ovarian cancer.

[0696] Panel 4D Summary: Ag1830 Highest expression of the CG5603 g-01 gene is seen in liver cirrhosis (CT=32.7). Furthermore, no expression in normal liver is seen in Panel 1.3D. Thus, the putative GPCR encoded for by this gene could potentially allow cells within the liver to respond to specific microenvironmental signals. Therefore, therapies designed with the protein encoded for by this gene may potentially modulate liver function and play a role in the identification and treatment of inflammatory or autoimmune diseases which effect the liver including liver cirrhosis and fibrosis.

[0697] In addition, expression of this gene could be used for the diagnosis of liver cirrhosis. Low but significant expression is also seen in a sample derived from a patient with IBD colitis, but not in normal colon. This suggests that the protein encoded by this gene may be involved in the disease process that results in inflammatory bowel disease. Therefore, therapeutic modulation of the expression or function of this gene product could potentially be useful in treating the symptoms of this disease. Please note that a second experiment with probe and primer set Ag1730 showed low/undetectable levels of expression in all the samples on this panel.(CTs>34.5) (Data not shown.) (Mark et al., G protein modulation of recombinant P/Q-type calcium channels by regulators of G protein signalling proteins. J. Physiol. 528 Pt 1:65-77, 2001).

[0698] AD. CG90341-02/GMAC006313_A_da1: Olfactory Receptor

[0699] Expression of gene CG90341-02 was assessed using the primer-probe sets Ag1567 and Ag1915, described in Tables ADA and ADB. Results of the RTQ-PCR runs are shown in Tables ADC and ADD.

138TABLE ADAProbe Name Ag1567StartSEQ IDPrimersSequencesLengthPostionNO:Forward5′-cagtgacaccagtctcaatgaa-3′22551310ProbeTET-5′-cttcatccagacagccacggtgttag-3′-TAMRA26596311Reverse5′-gaagccataagacaccgtgata-3′22623312

[0700]

139

TABLE ADB

Probe Name Ag1915

Start
SEQ ID

Position
NO:

Forward
5′-cagtgacaccagtctcaatgaa-3′
22
551
313

Probe
TET-5′-cttcatccagacagccacggtgttag-3′-TAMRA
26
596
314

Reverse
5′-gaagccataagacaccgtgata-3′
22
623
315

[0701]

140

TABLE ADC

Panel 1.3D

Rel. Exp. (%) Ag1567, Run

Rel. Exp. (%) Ag1567, Run

Tissue Name
165529542
Tissue Name
165529542

Liver adenocarcinoma
48.6
Kidney (fetal)
0.0

Pancreas
0.0
Renal ca. 786-0
27.7

Pancreatic ca. CAPAN 2
15.0
Renal ca. A498
17.8

Adrenal gland
8.8
Renal ca. RXF 393
17.0

Thyroid
15.3
Renal ca. ACHN
0.0

Salivary gland
28.1
Renal ca. UO-31
0.0

Pituitary gland
33.7
Renal ca. TK-10
0.0

Brain (fetal)
10.6
Liver
0.0

Brain (whole)
82.9
Liver (fetal)
7.6

Brain (amygdala)
58.2
Liver ca. (hepatoblast)
0.0

HepG2

Brain (cerebellum)
39.8
Lung
22.2

Brain (hippocampus)
71.2
Lung (fetal)
0.0

Brain (substantia nigra)
82.4
Lung ca. (small cell) LX-1
0.0

Brain (thalamus)
100.0
Lung ca. (small cell)
0.0

NCI-H69

Cerebral Cortex
0.0
Lung ca. (s.cell var.)
11.2

SHP-77

Spinal cord
0.0
Lung ca. (large cell)NCI-
4.7

H460

glio/astro U87-MG
15.9
Lung ca. (non-sm.cell)
14.8

A549

glio/astro U-118-MG
21.5
Lung ca. (non-s.cell)
0.0

NCI-H23

astrocytoma SW1783
0.0
Lung ca. (non-s.cell)
9.6

HOP-62

neuro*; met SK-N-AS
13.8
Lung ca. (non-s.cl) NCI-
0.0

H522

astrocytoma SF-539
12.4
Lung ca. (squam.) SW
10.7

900

astrocytoma SNB-75
13.9
Lung ca. (squam.) NCI-
0.0

H596

glioma SNB-19
12.2
Mammary gland
0.0

glioma U251
39.0
Breast ca.* (pl.ef) MCF-7
30.8

glioma SF-295
22.5
Breast ca.* (pl.ef) MDA-
0.0

MB-231

Heart (fetal)
0.0
Breast ca.* (pl.ef) T47D
0.0

Heart
0.0
Breast ca. BT-549
0.0

Skeletal muscle (fetal)
0.0
Breast ca. MDA-N
0.0

Skeletal muscle
0.0
Ovary
0.0

Bone marrow
46.3
Ovarian ca. OVCAR-3
32.5

Thymus
82.4
Ovarian ca. OVCAR-4
13.2

Spleen
31.9
Ovarian ca. OVCAR-5
0.0

Lymph node
99.3
Ovarian ca. OVCAR-8
0.0

Colorectal
42.3
Ovarian ca. IGROV-1
0.0

Stomach
42.9
Ovarian ca.* (ascites)
15.7

SK-OV-3

Small intestine
21.6
Uterus
35.1

Colon ca. SW480
0.0
Plancenta
24.1

Colon ca.* SW620(SW480
0.0
Prostate
11.3

met)

Colon ca. HT29
0.0
Prostate ca.* (bone
0.0

met)PC-3

Colon ca. HCT-116
0.0
Testis
0.0

Colon ca. CaCo-2
0.0
Melanoma Hs688(A).T
0.0

Colon ca.
20.9
Melanoma* (met)
0.0

tissue(ODO3866)

Hs688(B).T

Colon ca. HCC-2998
0.0
Melanoma UACC-62
0.0

Gastric ca.* (liver met)
14.9
Melanoma M14
11.1

NCI-N87

Bladder
9.5
Melanoma LOX IMVI
0.0

Trachea
7.9
Melanoma* (met) SK-
0.0

MEL-5

Kidney
9.2
Adipose
0.0

[0702]

141

TABLE ADD

Panel 4D

Rel. Exp. (%)
Rel. Exp. (%)

Rel. Exp. (%)
Rel. Exp. (%)

Ag1567, Run
Ag1915, Run

Ag1567, Run
Ag1915, Run

Tissue Name
165301511
160653158
Tissue Name
165301511
160653158

Secondary Th1 act
18.2
29.3
HUVEC IL-1beta
7.7
9.7

Secondary Th2 act
3.6
14.8
HUVEC IFN gamma
72.2
94.6

Secondary Tr1 act
24.0
24.5
HUVEC TNF alpha +
24.7
14.7

IFN gamma

Secondary Th1 rest
29.1
21.2
HUVEC TNF alpha +
0.0
15.6

IL4

Secondary Th2 rest
20.6
14.6
HUVEC IL-11
34.4
47.0

Secondary Tr1 rest
37.4
23.3
Lung Microvascular
49.7
61.6

EC none

Primary Th1 act
26.8
4.7
Lung Microvascular
58.2
33.0

EC TNF alpha + IL-

1beta

Primary Th2 act
5.3
18.4
Microvascular
91.4
57.0

Dermal EC none

Primary Tr1 act
29.7
25.2
Microsvasular Dermal
13.7
34.9

EC TNF alpha + IL-

1beta

Primary Th1 rest
93.3
58.2
Bronchial epithelium
32.3
5.2

TNF alpha + IL1beta

Primary Th2 rest
33.7
39.0
Small airway
8.0
15.3

epithelium none

Primary Tr1 rest
26.8
5.2
Small airway
39.5
39.2

epithelium TNF alpha +

IL-1beta

CD45RA CD4
5.8
15.7
Coronery artery SMC
0.0
0.0

lymphocyte act

rest

CD45RO CD4
22.1
3.7
Coronery artery SMC
9.1
0.0

lymphocyte act

TNF alpha + IL-1beta

CD8 lymphocyte act
13.7
10.7
Astrocytes rest
0.0
10.4

Secondary CD8
25.7
14.1
Astrocytes TMFalpha +
0.0
0.0

lymphocyte rest

IL-1beta

Secondary CD8
7.6
5.6
KU-812 (Basophil)
16.3
13.8

lymphocyte act

rest

CD4 lymphocyte
8.1
5.1
KU-812 (Basophil)
25.7
46.7

none

PMA/ionomycin

2ry
100.0
46.7
CCD1106
11.8
0.0

Th1/Th2/Tr1_anti-

(Keratinocytes) none

CD95 CH11

LAK cells rest
14.6
15.4
CCD1106
0.0
0.0

(Keratinocytes)

TNF alpha + IL-1beta

LAK cells IL-2
19.2
6.7
Liver cirrhosis
17.0
22.8

LAK cells IL-2 + IL-
28.7
44.1
Lupus kidney
15.4
0.0

12

LAK cells IL-2 + IFN
32.5
55.5
NCI-H292 none
18.8
17.0

gamma

LAK cells IL-2 + IL-
24.1
12.9
NCI-H292 IL-4
14.6
15.2

18

LAK cells
0.0
8.5
NCI-H292 IL-9
6.6
16.4

PMA/ionomycin

NK Cells IL-2 rest
4.0
11.5
NCI-H292 IL-13
0.0
8.8

Two Way MLR 3
33.0
11.1
NCI-H292 IFN
19.3
4.4

day

gamma

Two Way MLR 5
11.5
6.8
HPAEC none
37.9
59.0

day

Two Way MLR 7
9.8
13.9
HPAEC TNF alpha +
16.4
62.4

day

IL-1beta

PBMC rest
5.2
0.0
Lung fibroblast none
13.9
20.7

PBMC PWM
42.3
23.3
Lung fibroblast TNF
6.4
0.0

alpha + IL-1beta

PBMC PHA-L
0.0
7.6
Lung fibroblast IL-4
28.3
22.4

Ramos (B cell) none
28.5
24.1
Lung fibroblast IL-9
23.3
0.0

Ramos (B cell)
0.0
14.3
Lung fibroblast IL-13
23.3
6.3

ionomycin

B lymphocytes PWM
32.5
25.3
Lung fibroblast IFN
4.0
21.8

gamma

B lymphocytes
63.3
22.5
Dermal fibroblast
29.7
2.9

CD40L and IL-4

CCD1070 rest

EOL-1 dbcAMP
17.4
15.1
Dermal fibroblast
72.7
71.7

CCD1070 TNF alpha

EOL-1 dbcAMP
21.0
0.0
Dermal fibroblast
5.0
0.0

PMA/ionomycin

CCD1070 IL-1beta

Dendritic cells none
29.9
0.0
Dermal fibroblast IFN
12.8
17.9

gamma

Dendritic cells LPS
20.3
0.0
Dermal fibroblast IL-4
18.7
14.4

Dendritic cells anti-
15.0
25.9
IBD Colitis 2
7.1
0.0

CD40

Monocytes rest
8.4
14.3
IBD Crohn's
0.0
11.6

Monocytes LPS
11.3
0.0
Colon
36.6
6.4

Macrophages rest
24.5
22.5
Lung
38.4
20.7

Macrophages LPS
0.0
0.0
Thymus
34.4
21.5

HUVEC none
40.6
45.4
Kidney
60.3
59.9

HUVEC starved
53.6
100.0

[0703] Panel 1.3D Summary: Ag1567 This gene represents a novel G-protein coupled receptor (GPCR) with expression in the brain. The GPCR family of receptors contains a large number of neurotransmitter receptors, including the dopamine, serotonin, a and b-adrenergic, acetylcholine muscarinic, histamine, peptide, and metabotropic glutamate receptors. GPCRs are excellent drug targets in various neurologic and psychiatric diseases. All antipsychotics have been shown to act at the dopamine D2 receptor; similarly novel antipsychotics also act at the serotonergic receptor, and often the muscarinic and adrenergic receptors as well. While the majority of antidepressants can be classified as selective serotonin reuptake inhibitors, blockade of the 5-HT1A and a2 adrenergic receptors increases the effects of these drugs. The GPCRs are also of use as drug targets in the treatment of stroke. Blockade of the glutamate receptors may decrease the neuronal death resulting from excitotoxicity; further more the purinergic receptors have also been implicated as drug targets in the treatment of cerebral ischemia. The b-adrenergic receptors have been implicated in the treatment of ADHD with Ritalin, while the a-adrenergic receptors have been implicated in memory. Therefore this gene may be of use as a small molecule target for the treatment of any of the described diseases. Please note that a second experiment with the probe and primer set Ag1915 showed low/undetectable levels of expression in all samples on this panel. (CTs>34.5). (Data not shown.) (El Yacoubi et al., Adenosine A2A receptor antagonists are potential antidepressants: evidence based on pharmacology and A2A receptor knockout mice. Br J Pharmacol 134(1):68-77, 2001; Blier, Pharmacology of rapid-onset antidepressant treatment strategies. Clin Psychiatry 62 Suppl 15:12-7, 2001; Tranquillini and Reggiani, Glycine-site antagonists and stroke. Expert Opin Investig Drugs 8(11):1837-1848, 1999; Monopoli et al., Blockade of adenosine A2A receptors by SCH 58261 results in neuroprotective effects in cerebral ischaemia in rats. Neuroreport 9(17):3955-9, 1998).

[0704] Panel 2.2 Summary: Ag1567 Expression of the CG90341-02 gene is low/undetectable (CTs>35) across all of the samples on this panel (data not shown).

[0705] Panel 4D Summary: Ag1567/1915 Consistent but low expression of the CG90341-02 gene is observed in kidney, primary Th1 and Th2 cells and primary T cells stimulated with anti-CD95, as well as in HUVEC and HPAEC endothelial cells and dermal fibroblasts. In addition, expression is seen in dermal fibroblast cells stimulated with TNFalpha. The protein encoded by this transcript is a putative GPCR that may be important in T cell activation or polarization and dermal fibroblast response to proinflammatory cytokines. Therefore, therapies designed with the protein encoded by this transcript may be important in diseases mediated by T cells including asthma, IBD, or arthritis. Based on the dermal fibroblast expression seen in this panel, these therapies may also be important for the treatment of skin diseases including psoriasis and allergies.

[0706] AE. CG56385-01: Olfactory Receptor

[0707] Expression of gene CG56385-01 was assessed using the primer-probe sets Ag1833 and Ag1732, described in Tables AEA and AEB. Results of the RTQ-PCR runs are shown in Tables AEC, AED and AEE.

142TABLE AEAProbe Name Ag1833StartSEQ IDPrimersSequencesLengthPositionNO:Forward5′-gacaaaatggcatctgtgttct-3′22792316ProbeTET-5′-agtcattcccatgttgaatccactgg-3′-TAMRA26821317Reverse5′-tctttgttcctcaggctgtaga-3′22847318

[0708]

143

TABLE AEB

Probe Name Ag1732

Start
SEQ ID

Primers
Sequences
Length
Position
NO:

Forward
5′-gacaaaatggcatctgtgttct-3′
22
792
319

Probe
TET-5′-agtcattcccatgttgaatccactgg-3′-TAMRA
26
821
320

Reverse
5′-tctttgttcctcaggctgtaga-3′
22
847
321

[0709]

144

TABLE AEC

Panel 1.3D

Rel. Exp. (%) Ag1833, Run

Rel. Exp. (%) Ag1833, Run

Tissue Name
165975011
Tissue Name
165975011

Liver adenocarcinoma
0.0
Kidney (fetal)
0.0

Pancreas
25.9
Renal ca. 786-0
0.0

Pancreatic ca. CAPAN 2
0.0
Renal ca. A498
0.0

Adrenal gland
0.0
Renal ca. RXF 393
19.9

Thyroid
0.0
Renal ca. ACHN
0.0

Salivary gland
0.0
Renal ca. UO-31
0.0

Pituitary gland
0.0
Renal ca. TK-10
0.0

Brain (fetal)
0.0
Liver
0.0

Brain (whole)
0.0
Liver (fetal)
6.9

Brain (amygdala)
0.0
Liver ca. (hepatoblast)
0.0

HepG2

Brain (cerebellum)
0.0
Lung
0.0

Brain (hippocampus)
0.0
Lung (fetal)
0.0

Brain (substantia nigra)
0.0
Lung ca. (small cell) LX-1
0.0

Brain (thalamus)
25.9
Lung ca. (small cell)
0.0

NCI-H69

Cerebral Cortex
0.0
Lung ca. (s.cell var.)
0.0

SHP-77

Spinal cord
0.0
Lung ca. (large cell)NCI-
0.0

H460

glio/astro U87-MG
0.0
Lung ca. (non-sm.cell)
0.0

A549

glio/astro U-118-MG
0.0
Lung ca. (non-s.cell)
28.1

NCI-H23

astrocytoma SW1783
0.0
Lung ca. (non-s.cell)
0.0

HOP-62

neuro*; met SK-N-AS
0.0
Lung ca. (non-s.cl) NCI-
0.0

H522

astrocytoma SF-539
0.0
Lung ca. (squam.) SW
0.0

900

astrocytoma SNB-75
0.0
Lung ca. (squam.) NCI-
0.0

H596

glioma SNB-19
18.6
Mammary gland
0.0

glioma U251
0.0
Breast ca.* (pl.ef) MCF-7
0.0

glioma SF-295
0.0
Breast ca.* (pl.ef) MDA-
0.0

MB-231

Heart (Fetal)
0.0
Breast ca.* (pl.ef) T47D
0.0

Heart
0.0
Breast ca. BT-549
0.0

Skeletal muscle (Fetal)
0.0
Breast ca. MDA-N
0.0

Skeletal muscle
0.0
Ovary
0.0

Bone marrow
0.0
Ovarian ca. OVCAR-3
0.0

Thymus
0.0
Ovarian ca. OVCAR-4
0.0

Spleen
100.0
Ovarian ca. OVCAR-5
0.0

Lymph node
0.0
Ovarian ca. OVCAR-8
0.0

Colorectal
0.0
Ovarian ca. IGROV-1
0.0

Stomach
0.0
Ovarian ca. (ascites) SK-
66.4

OV-3

Small intestine
0.0
Uterus
0.0

Colon ca. SW480
0.0
Placenta
0.0

Colon ca.* SW620
0.0
Prostate
0.0

(SW480 met)

Colon ca. HT29
0.0
Prostate ca.* (bone met)
0.0

PC-3

Colon ca. HCT-116
0.0
Testis
0.0

Colon ca. CaCo-2
0.0
Melanoma Hs688(A).T
0.0

CC Well to Mod Diff
0.0
Melanoma* (met)
0.0

(ODO3866)

Hs688(B).T

Colon ca. HCC-2998
0.0
Melanoma UACC-62
0.0

Gastric ca. (liver met)
0.0
Melanoma M14
0.0

NCI-N87

Bladder
0.0
Melanoma LOX IMVI
0.0

Trachea
0.0
Melanoma* (met) SK-
0.0

MEL-5

Kidney
0.0
Adipose
0.0

[0710]

145

TABLE AED

Panel 2.2

Rel. Exp. (%)
Rel. Exp. (%)

Rel. Exp. (%)
Rel. Exp. (%)

Ag1732, Run
Ag1833, Run

Ag1732, Run
Ag1833, Run

Tissue Name
173761943
174229569
Tissue Name
173761943
174229569

Normal Colon
0.0
0.0
Kidney Margin
0.0
0.0

(OD04348)

Colon cancer
0.0
0.0
Kidney malignant
0.0
0.0

(OD06064)

cancer

(OD06204B)

Colon Margin
0.0
0.0
Kidney normal
0.0
0.0

(OD06064)

adjacent tissue

(OD06204E)

Colon cancer
0.0
0.0
Kidney Cancer
0.0
0.0

(OD06159)

(OD04450-01)

Colon Margin
0.0
0.0
Kidney Margin
0.0
0.0

(OD06159)

(OD04450-03)

Colon cancer
0.0
0.0
Kidney Cancer
0.0
0.0

(OD06297-04)

8120613

Colon Margin
0.0
0.0
Kidney Margin
0.0
0.0

(OD06297-015)

8120614

CC Gr.2 ascend
0.0
0.0
Kidney Cancer
0.0
0.0

colon (ODO3921)

9010320

CC Margin
0.0
0.0
Kidney Margin
0.0
0.0

(ODO3921)

9010321

Colon cancer
0.0
0.0
Kidney Cancer
0.0
0.0

metastasis

8120607

(OD06104)

Lung Margin
0.0
0.0
Kidney Margin
0.0
0.0

(OD06104)

8120608

Colon mets to lung
0.0
0.0
Normal Uterus
0.0
0.0

(OD04451-01)

Lung Margin
0.0
0.0
Uterine Cancer
0.0
0.0

(OD04451-02)

064011

Normal Prostate
0.0
0.0
Normal Thyroid
0.0
0.0

Prostate Cancer
3.2
0.0
Thyroid Cancer
0.0
0.0

(OD04410)

Prostate Margin
0.0
0.0
Thyroid Cancer
0.0
0.0

(OD04410)

A302152

Normal Ovary
0.0
0.0
Thyroid Margin
0.0
0.0

A302153

Ovarian cancer
0.0
0.0
Normal Breast
0.0
0.0

(OD06283-03)

Ovarian Margin
0.0
38.4
Breast Cancer
16.7
0.0

(OD06283-07)

Ovarian Cancer
100.0
100.0
Breast Cancer
12.2
0.0

Ovarian cancer
0.0
0.0
Breast Cancer
0.0
0.0

(OD06145)

(OD04590-01)

Ovarian Margin
0.0
0.0
Breast Cancer Mets
0.0
0.0

(OD06145)

(OD04590-03)

Ovarian cancer
0.0
0.0
Breast Cancer
0.0
0.0

(OD06455-03)

Metastasis

Ovarian Margin
0.0
0.0
Breast Cancer
0.0
0.0

(OD06455-07)

Normal Lung
0.0
0.0
Breast Cancer
0.0
0.0

9100266

Invasive poor diff.
0.0
0.0
Breast Margin
0.0
0.0

lung adeno

9100265

(ODO4945-01

Lung Margin
0.0
0.0
Breast Cancer
0.0
0.0

(ODO4945-03)

A209073

Lung Malignant
12.8
0.0
Breast Margin
0.0
0.0

Cancer (OD03126)

A2090734

Lung Margin
0.0
0.0
Breast cancer
0.0
0.0

(OD03126)

(OD06083)

Lung Cancer
0.0
0.0
Breast cancer node
0.0
0.0

(OD05014A)

metastasis

(OD06083)

Lung Margin
0.0
0.0
Normal Liver
0.0
0.0

(OD05014B)

Lung cancer
0.0
0.0
Liver Cancer 1026
0.0
0.0

(OD06081)

Lung Margin
0.0
0.0
Liver Cancer 1025
2.8
0.0

(OD06081)

Lung Cancer
0.0
0.0
Liver Cancer 6004-T
0.0
0.0

(OD04237-01)

Lung Margin
0.0
0.0
Liver Tissue 6004-N
0.0
0.0

(OD04237-02)

Ocular Mel Met to
0.0
0.0
Liver Cancer 6005-T
0.0
0.0

Liver (ODO4310)

Liver Margin
0.0
0.0
Liver Tissue 6005-N
0.0
0.0

(ODO4310)

Melanoma
0.0
0.0
Liver Cancer
0.0
0.0

Metastasis

Lung Margin
0.0
0.0
Normal Bladder
0.0
0.0

(OD04321)

Normal Kidney
0.0
0.0
Bladder Cancer
0.0
0.0

Kidney Ca, Nuclear
2.2
0.0
Bladder Cancer
0.0
0.0

grade 2 (OD04338)

Kidney Margin
0.0
0.0
Normal Stomach
0.0
0.0

(OD04338)

Kidney Ca Nuclear
0.0
0.0
Gastric Cancer
0.0
0.0

grade 1/2

9060397

(OD04339)

Kidney Margin
0.0
0.0
Stomach Margin
0.0
0.0

(OD04339)

9060396

Kidney Ca, Clear
0.0
0.0
Gastric Cancer
21.8
0.0

cell type

9060395

(OD04340)

Kidney Margin
0.0
0.0
Stomach Margin
0.0
0.0

(OD04340)

9060394

Kidney Ca, Nuclear
0.0
0.0
Gastric Cancer
0.0
0.0

grade 3 (OD04348)

064005

[0711]

146

TABLE AEE

Panel 4D

Rel. Exp. (%) Ag1833,

Rel. Exp. (%) Ag1833,

Tissue Name
Run 165824840
Tissue Name
Run 165824840

Secondary Th1 act
0.0
HUVEC IL-1 beta
0.0

Secondary Th2 act
0.0
HUVEC IFN gamma
0.0

Secondary Tr1 act
0.0
HUVEC TNF alpha + IFN
0.0

gamma

Secondary Th1 rest
0.0
HUVEC TNF alpha + IL4
0.0

Secondary Th2 rest
0.0
HUVEC IL-11
0.0

Secondary Tr1 rest
0.0
Lung Microvascular EC none
0.0

Primary Th1 act
0.0
Lung Microvascular EC
0.0

TNF alpha + IL-1 beta

Primary Th2 act
0.0
Microvascular Dermal EC none
0.0

Primary Tr1 act
0.0
Microsvasular Dermal EC
0.0

TNF alpha + IL-1 beta

Primary Th1 rest
0.0
Bronchial epithelium TNF alpha +
2.9

IL1 beta

Primary Th2 rest
0.0
Small airway epithelium none
0.0

Primary Tr1 rest
0.0
Small airway epithelium
0.0

TNF alpha + IL-1 beta

CD45RA CD4 lymphocyte
0.0
Coronery artery SMC rest
0.0

act

CD45RO CD4 lymphocyte
0.0
Coronery artery SMC TNF alpha +
0.0

act

IL-1 beta

CD8 lymphocyte act
0.0
Astrocytes rest
0.0

Secondary CD8
0.0
Astrocytes TNF alpha + IL-1 beta
0.0

lymphocyte rest

Secondary CD8
0.0
KU-812 (Basophil) rest
0.0

lymphocyte act

CD4 lymphocyte none
0.0
KU-812 (Basophil)
0.0

PMA/ionomycin

2ry Th1/Th2/Tr1_anti-
0.0
CCD1106 (Keratinocytes) none
0.0

CD95 CH11

LAK cells rest
2.6
CCD1106 (Keratinocytes)
0.0

TNF alpha + IL-1 beta

LAK cells IL-2
11.7
Liver cirrhosis
100.0

LAK cells IL-2 + IL-12
1.9
Lupus kidney
0.0

LAK cells IL-2 + IFN
0.0
NCI-H292 none
0.0

gamma

LAK cells IL-2 + IL-18
0.0
NCI-H292 IL-4
0.0

LAK cells
0.0
NCI-H292 IL-9
0.0

PMA/ionomycin

NK Cells IL-2 rest
0.0
NCI-H292 IL-13
0.0

Two Way MLR 3 day
2.7
NCI-H292 IFN gamma
0.0

Two Way MLR 5 day
0.0
HPAEC none
0.0

Two Way MLR 7 day
1.4
HPAEC TNF alpha + IL-1 beta
0.0

PBMC rest
0.0
Lung fibroblast none
0.0

PBMC PWM
0.0
Lung fibroblast TNF alpha + IL-
0.0

1 beta

PBMC PHA-L
0.0
Lung fibroblast IL-4
0.0

Ramos (B cell) none
0.0
Lung fibroblast IL-9
0.0

Ramos (B cell) ionomycin
0.0
Lung fibroblast IL-13
0.0

B lymphocytes PWM
0.0
Lung fibroblast IFN gamma
0.0

B lymphocytes CD40L
10.3
Dermal fibroblast CCD1070 rest
0.0

and IL-4

EOL-1 dbcAMP
0.0
Dermal fibroblast CCD1070
12.5

TNF alpha

EOL-1 dbcAMP
0.0
Dermal fibroblast CCD1070 IL-
0.0

PMA/ionomycin

1 beta

Dendritic cells none
0.0
Dermal fibroblast IFN gamma
0.0

Dendritic cells LPS
0.0
Dermal fibroblast IL-4
0.0

Dendritic cells anti-CD40
0.0
IBD Colitis 2
0.9

Monocytes rest
0.0
IBD Crohn's
0.0

Monocytes LPS
0.0
Colon
0.0

Macrophages rest
0.0
Lung
0.0

Macrophages LPS
1.3
Thymus
0.0

HUVEC none
0.0
Kidney
0.0

HUVEC starved
0.0

[0712] Panel 1.3D Summary: Ag1833 Expression of the CG56385-01 gene is highest (CT=33.9), in the spleen an important site of secondary immune responses. Therefore, antibodies or small molecule therapeutics that block the function of this GPCR may be usefull as anti-inflammatory therapeutics for the treatment of allergies, autoimmune diseases, and inflammatory diseases. In addition, low but significant expression of this gene is also detected in an ovarian cancer cell line (CT=34.5).

[0713] Data from a second experiment with Ag1732 is not included because the amp plot suggests that there were experimental difficulties with this run.

[0714] Panel 2.2Summary: Ag1732/Ag1833Results from two experiments using an identical probe/primer set are very comparable. Significant expression of the CG56385-01 gene is limited to a single ovarian cancer sample in this panel. These results are consistent with what was seen in Panel 1.3D. Therefore, expression of this gene may be used to distinguish ovarian cancers from the other samples on this panel. Furthermore, therapeutic modulation of the GPCR encoded by this gene may be beneficial in the treatment of ovarian cancer.

[0715] Panel 4D Summary: Ag1833 Significant expression of the CG56385-01 gene is detected in a liver cirrhosis sample (CT=32.5). Furthermore, expression of this gene is not detected in normal liver in Panel 1.3D, suggesting that its expression is unique to liver cirrhosis. This gene encodes a putative GPCR; therefore, antibodies or small molecule therapeutics could reduce or inhibit fibrosis that occurs in liver cirrhosis. In addition, antibodies to this putative GPCR could also be used for the diagnosis of liver cirrhosis. Data from a second experiment with Ag1732 shows low/undetectable expression in all the samples on this panel. (Data not shown.)

[0716] AF. CG56755-01/GMAC022882_D: Olfactory Receptor

[0717] Expression of gene CG56755-01 was assessed using the primer-probe set Ag1669, described in Table AFA. Results of the RTQ-PCR runs are shown in Tables AFB.

147TABLE AFAProbe Name Ag1669StartSEQ IDPrimersSequencesLengthPositionNO:Forward5′-agccatgtcctacgacctctat-3′22366322ProbeTET-5′-tggccatctgtaaccctctgctataca-3′-TAMRA27421323Reverse5′-gacatacccttcgtgacatgat-3′22421324

[0718]

148

TABLE AFB

Panel 4D

Rel. Exp. (%) Ag1669,

Rel. Exp. (%) Ag1669,

Tissue Name
Run 164729479
Tissue Name
Run 164729479

Secondary Th1 act
0.0
HUVEC IL-1 beta
0.0

Secondary Th2 act
0.0
HUVEC IFN gamma
0.0

Secondary Tr1 act
0.0
HUVEC TNF alpha + IFN
0.0

gamma

Secondary Th1 rest
0.0
HUVEC TNF alpha + IL4
0.0

Secondary Th2 rest
0.0
HUVEC IL-11
0.0

Secondary Tr1 rest
0.0
Lung Microvascular EC none
0.0

Primary Th1 act
0.0
Lung Microvascular EC
0.0

TNF alpha + IL-1 beta

Primary Th2 act
0.0
Microvascular Dermal EC none
0.0

Primary Tr1 act
0.0
Microsvasular Dermal EC
0.0

TNF alpha + IL-1 beta

Primary Th1 rest
0.0
Bronchial epithelium TNF alpha +
0.0

IL1 beta

Primary Th2 rest
0.0
Small airway epithelium none
0.0

Primary Tr1 rest
0.0
Small airway epithelium
0.0

TNF alpha + IL-1 beta

CD45RA CD4 lymphocyte
100.0
Coronery artery SMC rest
0.0

act

CD45RO CD4 lymphocyte
0.0
Coronery artery SMC TNF alpha +
0.0

act

IL-1 beta

CD8 lymphocyte act
0.0
Astrocytes rest
0.0

Secondary CD8
0.0
Astrocytes TNF alpha + IL-1 beta
0.0

lymphocyte rest

Secondary CD8
0.0
KU-812 (Basophil) rest
0.0

lymphocyte act

CD4 lymphocyte none
0.0
KU-812 (Basophil)
0.0

PMA/ionomycin

2ry Th1/Th2/Tr1 _anti-
0.0
CCD1106 (Keratinocytes) none
0.0

CD95 CH11

LAK cells rest
0.0
CCD1106 (Keratinocytes)
0.0

TNF alpha + IL-1 beta

LAK cells IL-2
0.0
Liver cirrhosis
0.0

LAK cells IL-2 + IL-12
0.0
Lupus kidney
0.0

LAK cells IL-2 + IFN
0.0
NCI-H292 none
0.0

gamma

LAK cells IL-2 + IL-18
0.0
NCI-H292 IL-4
0.0

LAK cells
0.0
NCI-H292 IL-9
15.2

PMA/ionomycin

NK Cells IL-2 rest
0.0
NCI-H292 IL-13
0.0

Two Way MLR 3 day
0.0
NCI-H292 IFN gamma
0.0

Two Way MLR 5 day
0.0
HPAEC none
0.0

Two Way MLR 7 day
0.0
HPAEC TNF alpha + IL-1 beta
0.0

PBMC rest
0.0
Lung fibroblast none
1.0

PBMC PWM
0.0
Lung fibroblast TNF alpha + IL-
0.0

1 beta

PBMC PHA-L
0.0
Lung fibroblast IL-4
0.0

Ramos (B cell) none
0.0
Lung fibroblast IL-9
0.0

Ramos (B cell) ionomycin
0.0
Lung fibroblast IL-13
0.0

B lymphocytes PWM
0.0
Lung fibroblast IFN gamma
0.0

B lymphocytes CD40L
0.0
Dermal fibroblast CCD1070 rest
0.0

and IL-4

EOL-1 dbcAMP
0.0
Dermal fibroblast CCD1070
0.0

TNF alpha

EOL-1 dbcAMP
0.0
Dermal fibroblast CCD1070 IL-
0.0

PMA/ionomycin

1 beta

Dendritic cells none
0.0
Dermal fibroblast IFN gamma
0.0

Dendritic cells LPS
0.0
Dermal fibroblast IL-4
0.0

Dendritic cells anti-CD40
0.0
IBD Colitis 2
0.0

Monocytes rest
0.0
IBD Crohn's
0.0

Monocytes LPS
0.0
Colon
0.0

Macrophages rest
0.0
Lung
0.0

Macrophages LPS
0.0
Thymus
0.0

HUVEC none
0.0
Kidney
0.0

HUVEC starved
0.0

[0719] Panel 1.3D Summary: Ag1669 Expression of the CG56755-01 gene is low/undetectable in all samples in this panel (CTs>35).

[0720] Panel 2.2 Summary: Ag1669 Expression of the CG56755-01 gene is low/undetectable in all samples in this panel (CTs>35).

[0721] Panel 4D Summary: Ag1669 The CG56755-01 transcript is only expressed in activated CD45RA+T cells(CT=33). These T cells are naive T cells that have been activated with CD3 and CD28. Thus, the putative GPCR encoded for by this transcript may be important in T cell activation or participate in the functions of these T cells once they are activated. For example, GPCRs act as chemokine receptors and play crucial roles in directing activated T cells to inflammed tissues. Therefore, therapeutics designed with the protein encoded by this transcript could be important in regulating T cell function and treating T cell mediated diseases such as asthma, arthritis, psoriasis, IBD, and lupus.

[0722] AG. CG56820-01: Olfactory Receptor

[0723] Expression of gene CG56820-01 was assessed using the primer-probe set Ag2602, described in Table AGA. Results of the RTQ-PCR runs are shown in Table AGB.

149TABLE AGAProbe Name Ag2602StartSEQ IDPrimersSequencesLengthPositionNO:Forward5′-gtgctgagaaatggcttatttg-3′22450325ProbeTET-5′-cactccagtgcctgtgcttgcag-3′-TAMRA23473326Reverse5′-tcaatttcattcttggagcaat-3′22508327

[0724]

150

TABLE AGB

Panel 4D

Rel. Exp. (%) Ag2602,

Rel. Exp. (%) Ag2602,

Tissue Name
Run 164216249
Tissue Name
Run 164216249

Secondary Th1 act
0.0
HUVEC IL-1 beta
0.0

Secondary Th2 act
5.1
HUVEC IFN gamma
0.0

Secondary Tr1 act
0.0
HUVEC TNF alpha + IFN
0.0

gamma

Secondary Th1 rest
6.8
HUVEC TNF alpha + IL4
1.8

Secondary Th2 rest
0.0
HUVEC IL-11
0.0

Secondary Tr1 rest
1.8
Lung Microvascular EC none
2.4

Primary Th1 act
2.6
Lung Microvascular EC
1.4

TNF alpha + IL-1 beta

Primary Th2 act
0.0
Microvascular Dermal EC none
0.0

Primary Tr1 act
0.0
Microsvasular Dermal EC
0.0

TNF alpha + IL-1 beta

Primary Th1 rest
49.7
Bronchial epithelium TNF alpha +
0.0

IL1 beta

Primary Th2 rest
23.2
Small airway epithelium none
0.0

Primary Tr1 rest
3.8
Small airway epithelium
1.2

TNF alpha + IL-1 beta

CD45RA CD4 lymphocyte
2.5
Coronery artery SMC rest
0.0

act

CD45RO CD4 lymphocyte
14.9
Coronery artery SMC TNF alpha +
0.0

act

IL-1 beta

CD8 lymphocyte act
3.2
Astrocytes rest
0.0

Secondary CD8
18.8
Astrocytes TNF alpha + IL-1 beta
0.0

lymphocyte rest

Secondary CD8
0.0
KU-812 (Basophil) rest
1.6

lymphocyte act

CD4 lymphocyte none
13.9
KU-812 (Basophil)
0.0

PMA/ionomycin

2ry Th1/Th2/Tr1_anti-
1.9
CCD1106 (Keratinocytes) none
0.0

CD95 CH11

LAK cells rest
11.5
CCD1106 (Keratinocytes)
0.0

TNF alpha + IL-1 beta

LAK cells IL-2
33.9
Liver cirrhosis
7.1

LAK cells IL-2 + IL-12
24.1
Lupus kidney
3.6

LAK cells IL-2 + IFN
100.0
NCI-H292 none
1.4

gamma

LAK cells IL-2 + IL-18
34.2
NCI-H292 IL-4
1.3

LAK cells
4.0
NCI-H292 IL-9
0.0

PMA/ionomycin

NK Cells IL-2 rest
11.2
NCI-H292 IL-13
0.0

Two Way MLR 3 day
32.1
NCI-H292 IFN gamma
2.4

Two Way MLR 5 day
2.1
HPAEC none
0.0

Two Way MLR 7 day
3.3
HPAEC TNF alpha + IL-1 beta
0.0

PBMC rest
1.3
Lung fibroblast none
2.1

PBMC PWM
25.0
Lung fibroblast TNF alpha + IL-
0.0

1 beta

PBMC PHA-L
14.6
Lung fibroblast IL-4
1.2

Ramos (B cell) none
0.0
Lung fibroblast IL-9
3.5

Ramos (B cell) ionomycin
0.0
Lung fibroblast IL-13
0.0

B lymphocytes PWM
27.9
Lung fibroblast IFN gamma
4.0

B lymphocytes CD40L
7.3
Dermal fibroblast CCD1070 rest
0.0

and IL-4

EOL-1 dbcAMP
1.4
Dermal fibroblast CCD1070
12.2

TNF alpha

EOL-1 dbcAMP
0.0
Dermal fibroblast CCD1070 IL-
0.0

PMA/ionomycin

1 beta

Dendritic cells none
3.1
Dermal fibroblast IFN gamma
2.7

Dendritic cells LPS
3.0
Dermal fibroblast IL-4
0.0

Dendritic cells anti-CD40
1.5
IBD Colitis 2
1.5

Monocytes rest
1.7
IBD Crohn's
0.0

Monocytes LPS
1.8
Colon
11.7

Macrophages rest
6.0
Lung
4.8

Macrophages LPS
0.0
Thymus
3.6

HUVEC none
1.3
Kidney
16.7

HUVEC starved
0.0

[0725] CNS_neurodegeneration_v1.0 Summary: Ag2602 Expression of the CG56820-01 gene is low/undetectable (CTs>35) across all of the samples on this panel (data not shown).

[0726] Panel 1.3D Summary: Ag2602 Expression of the CG56820-01 gene is low/undetectable (CTs>35) across all of the samples on this panel (data not shown).

[0727] Panel 2.2 Summary: Ag2602 Expression of the CG56820-01 gene is low/undetectable (CTs>35) across all of the samples on this panel (data not shown).

[0728] Panel 4D Summary: Ag2602 Highest expression of the CG56820-01 transcript are detected in stimulated lymphokine-activated killer cells LAK)(CT=33.4). These cells are involved in tumor immunology and cell clearance of virally and bacterial infected cells as well as tumors. Therefore, modulation of the function of the protein encoded by this gene through the application of a small molecule drug or antibody may alter the functions of these cells and lead to improvement of symptoms associated with these conditions. Low levels of expression were also detected in a wide range of other cell types of significance in the immune response in health and disease.

[0729] AH. CG56799-01: Olfactory Receptor

[0730] Expression of gene CG56799-01 was assessed using the primer-probe set Ag2603, described in Table AHA. Results of the RTQ-PCR runs are shown in Tables AHB, AHC, AHD and AHE.

151TABLE AHAProbe Name Ag2603StartSEQ IDPrimersSequencesLengthPositionNO:Forward5′-tgtactacttcttggccatgct-3′22176328ProbeTET-5′-tagtacaatccctaaagccctctgca-3′-TAMRA26225329Reverse5′-tccttgagatgaaaccagaaga-3′22251330

[0731]

152

TABLE AHB

CNS_neurodegeneration_v1.0

Rel. Exp. (%) Ag2603, Run

Rel. Exp. (%) Ag2603, Run

Tissue Name
208779994
Tissue Name
208779994

AD 1 Hippo
0.0
Control (Path) 3
3.6

Temporal Ctx

AD 2 Hippo
7.9
Control (Path) 4
26.8

Temporal Ctx

AD 3 Hippo
0.0
AD 1 Occipital Ctx
3.1

AD 4 Hippo
3.4
AD 2 Occipital Ctx
0.0

(Missing)

AD 5 Hippo
0.0
AD 3 Occipital Ctx
0.0

AD 6 Hippo
57.4
AD 4 Occipital Ctx
11.5

Control 2 Hippo
13.5
AD 5 Occipital Ctx
16.8

Control 4 Hippo
1.9
AD 5 Occipital Ctx
0.0

Control (Path) 3 Hippo
3.6
Control 1 Occipital Ctx
0.0

AD 1 Temporal Ctx
7.5
Control 2 Occipital Ctx
7.1

AD 2 Temporal Ctx
2.1
Control 3 Occipital Ctx
7.1

AD 3 Temporal Ctx
3.2
Control 4 Occipital Ctx
2.8

AD 4 Temporal Ctx
5.5
Control (Path) 1
30.4

Occipital Ctx

AD 5 Inf Temporal Ctx
33.0
Control (Path) 2
3.1

Occipital Ctx

AD 5 Sup Temporal
26.8
Control (Path) 3
8.7

Ctx

Occipital Ctx

AD 6 lnf Temporal Ctx
100.0
Control (Path) 4
3.7

Occipital Ctx

AD 6 Sup Temporal
54.0
Control 1 Parietal Ctx
0.0

Ctx

Control 1 Temporal Ctx
0.0
Control 2 Parietal Ctx
3.8

Control 2 Temporal Ctx
17.0
Control 3 Parietal Ctx
16.3

Control 3 Temporal Ctx
9.6
Control (Path) 1
17.4

Parietal Ctx

Control 3 Temporal Ctx
0.0
Control (Path) 2
3.8

Parietal Ctx

Control (Path) 1
32.1
Control (Path) 3
6.5

Temporal Ctx

Parietal Ctx

Control (Path) 2
11.0
Control (Path) 4
13.3

Temporal Ctx

Parietal Ctx

[0732]

153

TABLE AHC

Panel 1.3D

Rel. Exp. (%) Ag2603, Run

Rel. Exp. (%) Ag2603, Run

Tissue Name
166219800
Tissue Name
166219800

Liver adenocarcinoma
0.0
Kidney (fetal)
0.0

Pancreas
6.8
Renal ca. 786-0
0.0

Pancreatic ca. CAPAN 2
0.0
Renal ca. A498
0.0

Adrenal gland
6.7
Renal ca. RXF 393
0.0

Thyroid
0.0
Renal ca. ACHN
0.0

Salivary gland
11.0
Renal ca. UO-31
8.0

Pituitary gland
6.5
Renal ca. TK-10
0.0

Brain (fetal)
12.9
Liver
0.0

Brain (whole)
7.3
Liver (fetal)
0.0

Brain (amygdala)
0.0
Liver ca. (hepatoblast)
0.0

HepG2

Brain (cerebellum)
6.9
Lung
62.9

Brain (hippocampus)
7.4
Lung (fetal)
13.5

Brain (substantia nigra)
0.0
Lung ca. (small cell) LX-1
0.0

Brain (thalamus)
0.0
Lung ca. (small cell)
0.0

NCI-H69

Cerebral Cortex
0.0
Lung ca. (s.cell var.)
0.0

SHP-77

Spinal cord
32.1
Lung ca. (large cell)NCI-
0.0

H460

glio/astro U87-MG
0.0
Lung ca. (non-sm. cell)
8.5

A549

glio/astro U-118-MG
0.0
Lung ca. (non-s.cell)
0.0

NCI-H23

astrocytoma SW1783
0.0
Lung ca. (non-s.cell)
5.9

HOP-62

neuro*; met SK-N-AS
0.0
Lung ca. (non-s.cl) NCI-
0.0

H522

astrocytoma SF-539
8.8
Lung ca. (squam.) SW
0.0

900

astrocytoma SNB-75
0.0
Lung ca. (squam.) NCI-
0.0

H596

glioma SNB-19
0.0
Mammary gland
30.8

glioma U251
8.8
Breast ca.* (pl.ef) MCF-7
0.0

glioma SF-295
0.0
Breast ca.* (pl.ef) MDA-
0.0

MB-231

Heart (Fetal)
0.0
Breast ca.* (pl.ef) T47D
15.0

Heart
17.6
Breast ca. BT-549
0.0

Skeletal muscle (Fetal)
0.0
Breast ca. MDA-N
0.0

Skeletal muscle
0.0
Ovary
0.0

Bone marrow
23.0
Ovarian ca. OVCAR-3
0.0

Thymus
44.1
Ovarian ca. OVCAR-4
0.0

Spleen
8.5
Ovarian ca. OVCAR-5
0.0

Lymph node
74.2
Ovarian ca. OVCAR-8
0.0

Colorectal
50.3
Ovarian ca. IGROV-1
4.7

Stomach
0.0
Ovarian ca. (ascites) SK-
100.0

OV-3

Small intestine
29.9
Uterus
2.2

Colon ca. SW480
0.0
Placenta
34.9

Colon ca.* SW620
0.0
Prostate
18.8

(SW480 met)

Colon ca. HT29
0.0
Prostate ca.* (bone met)
0.0

PC-3

Colon ca. HCT-116
0.0
Testis
20.2

Colon ca. CaCo-2
0.0
Melanoma Hs688(A).T
0.0

CC Well to Mod Diff
0.0
Melanoma* (met)
0.0

(ODO3866)

Hs688(B).T

Colon ca. HCC-2998
0.0
Melanoma UACC-62
8.9

Gastric ca. (liver met)
0.0
Melanoma M14
0.0

NCI-N87

Bladder
47.0
Melanoma LOX IMVI
0.0

Trachea
15.4
Melanoma* (met) SK-
0.0

MEL-5

Kidney
8.5
Adipose
31.9

[0733]

154

TABLE AHD

Panel 2.2

Rel. Exp. (%) Ag2603,

Rel. Exp. (%) Ag2603,

Tissue Name
Run 175127862
Tissue Name
Run 175127862

Normal Colon
3.5
Kidney Margin (OD04348)
100.0

Colon cancer (OD06064)
4.9
Kidney malignant cancer
0.0

(OD06204B)

Colon Margin (OD06064)
2.9
Kidney normal adjacent
7.7

tissue (OD06204E)

Colon cancer (OD06159)
0.0
Kidney Cancer (OD04450-
0.0

01)

Colon Margin (OD06159)
11.9
Kidney Margin (OD04450-
10.9

03)

Colon cancer (OD06297-04)
0.0
Kidney Cancer 8120613
0.0

Colon Margin (OD06297-
4.2
Kidney Margin 8120614
0.0

015)

CC Gr.2 ascend colon
0.0
Kidney Cancer 9010320
2.9

(ODO3921)

CC Margin (ODO3921)
5.0
Kidney Margin 9010321
0.0

Colon cancer metastasis
0.0
Kidney Cancer 8120607
0.0

(OD06104)

Lung Margin (OD06104)
9.8
Kidney Margin 8120608
2.7

Colon mets to lung
0.0
Normal Uterus
4.7

(OD04451-01)

Lung Margin (OD04451-02)
24.8
Uterine Cancer 064011
5.4

Normal Prostate
5.5
Normal Thyroid
4.1

Prostate Cancer (OD04410)
3.2
Thyroid Cancer
0.0

Prostate Margin (OD04410)
0.0
Thyroid Cancer A302152
0.0

Normal Ovary
0.0
Thyroid Margin A302153
0.0

Ovarian cancer (OD06283-
2.2
Normal Breast
26.8

03)

Ovarian Margin (OD06283-
27.9
Breast Cancer
7.5

07)

Ovarian Cancer
10.1
Breast Cancer
3.4

Ovarian cancer (OD06145)
0.0
Breast Cancer (OD04590-
0.0

01)

Ovarian Margin (OD06145)
13.9
Breast Cancer Mets
4.0

(OD04590-03)

Ovarian cancer (OD06455-
0.0
Breast Cancer Metastasis
0.0

03)

Ovarian Margin (OD06455-
22.2
Breast Cancer
0.0

07)

Normal Lung
9.0
Breast Cancer 9100266
3.1

Invasive poor diff. lung
19.9
Breast Margin 9100265
3.2

adeno (ODO4945-01

Lung Margin (ODO4945-03)
65.5
Breast Cancer A209073
0.0

Lung Malignant Cancer
0.0
Breast Margin A2090734
4.0

(OD03126)

Lung Margin (OD03126)
1.4
Breast cancer (OD06083)
8.0

Lung Cancer (OD05014A)
4.8
Breast cancer node
11.6

metastasis (OD06083)

Lung Margin (OD05014B)
7.9
Normal Liver
0.0

Lung cancer (OD06081)
3.0
Liver Cancer 1026
0.0

Lung Margin (OD06081)
3.1
Liver Cancer 1025
11.4

Lung Cancer (OD04237-01)
0.0
Liver Cancer 6004-T
11.0

Lung Margin (OD04237-02)
20.2
Liver Tissue 6004-N
5.8

Ocular Mel Met to Liver
0.0
Liver Cancer 6005-T
0.0

(ODO4310)

Liver Margin (ODO4310)
0.0
Liver Tissue 6005-N
7.5

Melanoma Metastasis
0.0
Liver Cancer
18.0

Lung Margin (OD04321)
4.5
Normal Bladder
8.5

Normal Kidney
0.0
Bladder Cancer
0.0

Kidney Ca, Nuclear grade 2
4.4
Bladder Cancer
22.5

(OD04338)

Kidney Margin (OD04338)
5.0
Normal Stomach
21.3

Kidney Ca Nuclear grade 1/2
0.0
Gastric Cancer 9060397
5.4

(OD04339)

Kidney Margin (OD04339)
0.0
Stomach Margin 9060396
5.7

Kidney Ca, Clear cell type
8.4
Gastric Cancer 9060395
7.9

(OD04340)

Kidney Margin (OD04340)
8.7
Stomach Margin 9060394
4.5

Kidney Ca, Nuclear grade 3
15.1
Gastric Cancer 064005
13.7

(OD04348)

[0734]

155

TABLE AHE

Panel 4D

Rel. Exp. (%) Ag2603,

Rel. Exp. (%) Ag2603,

Tissue Name
Run 164160188
Tissue Name
Run 164160188

Secondary Th1 act
0.0
HUVEC IL-1 beta
1.5

Secondary Th2 act
5.7
HUVEC IFN gamma
2.8

Secondary Tr1 act
0.0
HUVEC TNF alpha + IFN
0.0

gamma

Secondary Th1 rest
10.4
HUVEC TNF alpha + IL4
0.0

Secondary Th2 rest
3.1
HUVEC IL-11
0.0

Secondary Tr1 rest
9.6
Lung Microvascular EC none
1.5

Primary Th1 act
1.9
Lung Microvascular EC
0.0

TNF alpha + IL-1 beta

Primary Th2 act
5.3
Microvascular Dermal EC none
0.0

Primary Tr1 act
10.2
Microsvasular Dermal EC
0.0

TNF alpha + IL-1 beta

Primary Th1 rest
65.1
Bronchial epithelium TNF alpha +
6.5

IL1 beta

Primary Th2 rest
13.7
Small airway epithelium none
0.0

Primary Tr1 rest
12.3
Small airway epithelium
7.4

TNF alpha + IL-1 beta

CD45RA CD4 lymphocyte
15.8
Coronery artery SMC rest
0.0

act

CD45RO CD4 lymphocyte
29.9
Coronery artery SMC TNF alpha +
0.0

act

IL-1 beta

CD8 lymphocyte act
6.3
Astrocytes rest
0.0

Secondary CD8
42.9
Astrocytes TNF alpha + IL-1 beta
0.0

lymphocyte rest

Secondary CD8
2.4
KU-812 (Basophil) rest
0.0

lymphocyte act

CD4 lymphocyte none
35.1
KU-812 (Basophil)
1.0

PMA/ionomycin

2ry Th1/Th2/Tr1_anti-
14.2
CCD1106 (Keratinocytes) none
0.0

CD95 CH11

LAK cells rest
32.1
CCD1106 (Keratinocytes)
1.3

TNF alpha + IL-1 beta

LAK cells IL-2
80.7
Liver cirrhosis
17.4

LAK cells IL-2 + IL-12
81.2
Lupus kidney
3.1

LAK cells IL-2 + IFN
100.0
NCI-H292 none
1.4

gamma

LAK cells IL-2 + IL-18
95.9
NCI-H292 IL-4
0.7

LAK cells
6.7
NCI-H292 IL-9
2.1

PMA/ionomycin

NK Cells IL-2 rest
32.8
NCI-H292 IL-13
0.0

Two Way MLR 3 day
66.0
NCI-H292 IFN gamma
1.1

Two Way MLR 5 day
19.9
HPAEC none
0.0

Two Way MLR 7 day
10.1
HPAEC TNF alpha + IL-1 beta
0.0

PBMC rest
5.9
Lung fibroblast none
0.0

PBMC PWM
66.4
Lung fibroblast TNF alpha + IL-
0.9

1 beta

PBMC PHA-L
9.9
Lung fibroblast IL-4
0.3

Ramos (B cell) none
0.0
Lung fibroblast IL-9
2.8

Ramos (B cell) ionomycin
0.0
Lung fibroblast IL-13
1.0

B lymphocytes PWM
9.2
Lung fibroblast IFN gamma
0.0

B lymphocytes CD40L
9.7
Dermal fibroblast CCD1070 rest
0.0

and IL-4

EOL-1 dbcAMP
3.1
Dermal fibroblast CCD1070
0.0

TNF alpha

EOL-1 dbcAMP
0.0
Dermal fibroblast CCD1070 IL-
0.0

PMA/ionomycin

1 beta

Dendritic cells none
16.5
Dermal fibroblast IFN gamma
0.0

Dendritic cells LPS
6.5
Dermal fibroblast IL-4
6.1

Dendritic cells anti-CD40
3.0
IBD Colitis 2
6.0

Monocytes rest
5.4
IBD Crohn's
1.0

Monocytes LPS
3.2
Colon
15.2

Macrophages rest
18.2
Lung
5.3

Macrophages LPS
10.4
Thymus
8.4

HUVEC none
0.0
Kidney
52.5

HUVEC starved
0.0

[0735] CNS_neurodegeneration_v1.0 Summary: Ag2603 This gene represents a novel G-protein coupled receptor (GPCR) with expression in the brain. The GPCR family of receptors contains a large number of neurotransmitter receptors, including the dopamine, serotonin, a and b-adrenergic, acetylcholine muscarinic, histamine, peptide, and metabotropic glutamate receptors. GPCRs are excellent drug targets in various neurologic and psychiatric diseases. All antipsychotics have been shown to act at the dopamine D2 receptor; similarly novel antipsychotics also act at the serotonergic receptor, and often the muscarinic and adrenergic receptors as well. While the majority of antidepressants can be classified as selective serotonin reuptake inhibitors, blockade of the 5-HT1A and a2 adrenergic receptors increases the effects of these drugs. The GPCRs are also of use as drug targets in the treatment of stroke. Blockade of the glutamate receptors may decrease the neuronal death resulting from excitotoxicity; further more the purinergic receptors have also been implicated as drug targets in the treatment of cerebral ischemia. The b-adrenergic receptors have been implicated in the treatment of ADHD with Ritalin, while the a-adrenergic receptors have been implicated in memory. Therefore this gene may be of use as a small molecule target for the treatment of any of the described diseases. Furthermore, the expression profile of this GPCR is found to be upregulated in the temporal cortex of Alzheimer's disease patients. Thus, blockade of this receptor may be of use in the treatment of this disease and decrease neuronal death (El Yacoubi et al., Adenosine A2A receptor antagonists are potential antidepressants: evidence based on pharmacology and A2A receptor knockout mice. Br J Pharmacol 134(1):68-77, 2001; Blier, Pharmacology of rapid-onset antidepressant treatment strategies. Clin Psychiatry 62 Suppl 15:12-7, 2001; Tranquillini and Reggiani, Glycine-site antagonists and stroke. Expert Opin Investig Drugs 8(11):1837-1848, 1999; Monopoli et al., Blockade of adenosine A2A receptors by SCH 58261 results in neuroprotective effects in cerebral ischaemia in rats. Neuroreport 9(17):3955-9, 1998).

[0736] Panel 1.3D Summary: Ag2603 Expression of the CG56799-01 gene is highest in an ovarian cancer cell line (CT=33.6). Low but significant expression is also detected in lymph node, lung, colon and bladder. Therefore, expression of this gene may be used to distinguish these tissues from the other samples on this panel.

[0737] Panel 2.2 Summary: Ag2603 Highest expression of the gene in the CG56799-01 panel is seen in a sample derived from normal kidney adjacent to a tumor (CT=32.9). Significant expression is also seen in normal lung tissue adjacent to a tissue. Thus, expression of this gene could be used to differentiate these tissues from other samples on this panel.

[0738] Panel 4D Summary: Ag2603 The CG56799-01 gene is expressed at moderate levels to low levels in a wide range of cell types of significance in the immune response in health and disease and in normal tissues. Highest expression is seen in stimulated lymphokine-activated killer cells (LAK)(CT=30.1). These cells are involved in tumor immunology and cell clearance of tumors and virally and bacterial infected cells. Therefore, modulation of the function of this gene product with a small molecule drug or antibody may alter the functions of these cells and lead to improvement of symptoms associated with these conditions.

[0739] Low level expression is also detected in a wide range of other cell types of significance in the immune response in health and disease. Therefore, modulation of the function of this gene product with a small molecule drug or antibody may alter the functions of B cells, cells of the T-cell lineage, macrophages and monocytes and lead to improvement of the symptoms of patients suffering from autoimmune and inflammatory diseases such as asthma, allergies, inflammatory bowel disease, lupus erythematosus, arthritis, and cancer-related conditions.

[0740] AI. CG56929-01: Olfactory Receptor

[0741] Expression of gene CG56929-01 was assessed using the primer-probe set Ag2207, described in Table AIA. Results of the RTQ-PCR runs are shown in Table AIB.

156TABLE AIAProbe Name Ag2207StartSEQ IDPrimersSequencesLengthPositionNO:Forward5′-gctttggaaagcatcagataac-3′2257331ProbeTET-5′-cctctttgtggttttcctaactgtctaca-3′-TAMRA2979332Reverse5′-acaatgatgatgttagcaacca-3′22117333

[0742]

157

TABLE AIB

Panel 2.2

Rel. Exp. (%) Ag2207,

Rel. Exp. (%) Ag2207,

Tissue Name
Run 174166982

Run 174166982

Normal Colon
0.0
Kidney Margin (OD04348)
0.0

Colon cancer (OD06064)
0.0
Kidney malignant cancer
5.9

(OD06204B)

Colon Margin (OD06064)
0.0
Kidney normal adjacent
0.0

tissue (OD06204E)

Colon cancer (OD06159)
0.0
Kidney Cancer (OD04450-
0.0

01)

Colon Margin (OD06159)
0.0
Kidney Margin (OD04450-
0.0

03)

Colon cancer (OD06297-04)
0.0
Kidney Cancer 8120613
0.0

Colon Margin (OD06297-
0.0
Kidney Margin 8120614
0.0

015)

CC Gr.2 ascend colon
0.0
Kidney Cancer 9010320
0.0

(OD03921)

CC Margin (ODO3921)
0.0
Kidney Margin 9010321
0.0

Colon cancer metastasis
0.0
Kidney Cancer 8120607
0.0

(OD06104)

Lung Margin (OD06104)
0.0
Kidney Margin 8120608
0.0

Colon mets to lung
0.0
Normal Uterus
0.0

(OD04451-01)

Lung Margin (OD04451-02)
0.0
Uterine Cancer 064011
0.0

Normal Prostate
0.0
Normal Thyroid
0.0

Prostate Cancer (OD04410)
0.0
Thyroid Cancer 064010
6.8

Prostate Margin (OD04410)
0.0
Thyroid Cancer A302152
0.0

Normal Ovary
0.0
Thyroid Margin A302153
0.0

Ovarian cancer (OD06283-
0.0
Normal Breast
5.8

03)

Ovarian Margin (OD06283-
0.0
Breast Cancer (OD04566)
0.0

07)

Ovarian Cancer 064008
28.3
Breast Cancer 1024
0.0

Ovarian cancer (OD06145)
0.0
Breast Cancer (OD04590-
0.0

01)

Ovarian Margin (OD06145)
0.0
Breast Cancer Mets
0.0

(OD04590-03)

Ovarian cancer (OD06455-
0.0
Breast Cancer Metastasis
0.0

03)

(OD04655-05)

Ovarian Margin (OD06455-
0.0
Breast Cancer 064006
5.3

07)

Normal Lung
0.0
Breast Cancer 9100266
0.0

Invasive poor diff. lung
11.5
Breast Margin 9100265
0.0

adeno (ODO4945-01

Lung Margin (ODO4945-03)
0.0
Breast Cancer A209073
8.8

Lung Malignant Cancer
0.0
Breast Margin A2090734
0.0

(OD03126)

Lung Margin (OD03126)
0.0
Breast cancer (OD06083)
11.9

Lung Cancer (OD05014A)
6.0
Breast cancer node
0.0

metastasis (OD06083)

Lung Margin (OD05014B)
8.9
Normal Liver
100.0

Lung cancer (OD06081)
5.1
Liver Cancer 1026
5.9

Lung Margin (OD06081)
0.0
Liver Cancer 1025
20.6

Lung Cancer (OD04237-01)
0.0
Liver Cancer 6004-T
30.6

Lung Margin (OD04237-02)
0.0
Liver Tissue 6004-N
0.0

Ocular Melanoma Metastasis
0.0
Liver Cancer 6005-T
0.0

Ocular Melanoma Margin
12.0
Liver Tissue 6005-N
0.0

(Liver)

Melanoma Metastasis
0.0
Liver Cancer 064003
16.0

Melanoma Margin (Lung)
0.0
Normal Bladder
0.0

Normal Kidney
0.0
Bladder Cancer 1023
0.0

Kidney Ca, Nuclear grade 2
0.0
Bladder Cancer A302173
0.0

(OD04338)

Kidney Margin (OD04338)
0.0
Normal Stomach
0.0

Kidney Ca Nuclear grade 1/2
0.0
Gastric Cancer 9060397
5.8

(OD04339)

Kidney Margin (OD04339)
9.6
Stomach Margin 9060396
0.0

Kidney Ca, Clear cell type
0.0
Gastric Cancer 9060395
10.4

(OD04340)

Kidney Margin (OD04340)
0.0
Stomach Margin 9060394
4.8

Kidney Ca, Nuclear grade 3
0.0
Gastric Cancer 064005
0.0

(OD04348)

[0743] CNS_neurodegeneration_v1.0 Summary: Ag2207 Expression of the CG56929-01 gene is low/undetectable in all samples in this panel (CTs>35). (Data not shown.)

[0744] Panel 1.3D Summary: Ag2207 Expression of the CG56929-01 gene is low/undetectable in all samples in this panel (CTs>35). (Data not shown.)

[0745] Panel 2.2 Summary: Ag2207 Expression of the CG56929-01 gene is restricted to normal liver (CT=34.1). Thus, expression of this gene could be used to differentiate between normal liver and other samples on this panel and as a marker to detect the presence of liver cancer. Furthermore, therapeutic inhibition of the function or expression of the protein encoded by this gene may be useful in the treatment of liver cancer.

[0746] Panel 4D Summary: Ag2207 Expression of the CG56929-01 gene is low/undetectable in all samples in this panel (CTs>35). (Data not shown.)

[0747] AJ. CG57376-01 and GMAL049739_B: Olfactory Receptor

[0748] Expression of gene GMAL049739_B and full length physical clone CG57376-01 was assessed using the primer-probe set Ag2625, described in Table AJA. Results of the RTQ-PCR runs are shown in Table AJB.

158TABLE AJAProbe Name Ag2625StartSEQ IDPrimersSequencesLengthPositionNO:Forward5′-ccttctgggcttctctgaac-3′2036334ProbeTET-5′-cccagcactggaaaggactctctttg-3′2657335Reverse5′-ccagggtcaagaggtaggaa-3′2096336

[0749]

159

TABLE AJB

Panel 1.3D

Rel. Exp. (%) Ag2625, Run

Rel. Exp. (%) Ag2625, Run

Tissue Name
167649297
Tissue Name
167649297

Liver adenocarcinoma
0.0
Kidney (fetal)
0.0

Pancreas
0.0
Renal ca. 786-0
0.0

Pancreatic ca. CAPAN 2
0.0
Renal ca. A498
0.0

Adrenal gland
0.0
Renal ca. RXF 393
0.0

Thyroid
0.0
Renal ca. ACHN
0.0

Salivary gland
0.0
Renal ca. UO-31
0.0

Pituitary gland
0.0
Renal ca. TK-10
100.0

Brain (fetal)
0.0
Liver
0.0

Brain (whole)
0.0
Liver (fetal)
0.0

Brain (amygdala)
0.0
Liver ca. (hepatoblast)
0.0

HepG2

Brain (cerebellum)
0.0
Lung
0.0

Brain (hippocampus)
0.0
Lung (fetal)
0.0

Brain (substantia nigra)
2.4
Lung ca. (small cell) LX-1
0.0

Brain (thalamus)
0.0
Lung ca. (small cell)
0.0

NCI-H69

Cerebral Cortex
0.0
Lung ca. (s.cell var.)
0.0

SHP-77

Spinal cord
0.0
Lung ca. (large cell)NCI-
0.0

H460

glio/astro U87-MG
0.0
Lung ca. (non-sm.cell)
0.0

A549

glio/astro U-118-MG
0.0
Lung ca. (non-s.cell)
0.0

NCI-H23

astrocytoma SW1783
0.0
Lung ca. (non-s.cell)
0.0

HOP-62

neuro*; met SK-N-AS
0.0
Lung ca. (non-s.cl) NCI-
0.0

H522

astrocytoma SF-539
0.0
Lung ca. (squam.) SW
0.0

900

astrocytoma SNB-75
0.0
Lung ca. (squam.) NCI-
0.0

H596

glioma SNB-19
0.0
Mammary gland
0.0

glioma U251
1.3
Breast ca.* (pl.ef) MCF-7
0.0

glioma SF-295
0.0
Breast ca.* (pl.ef) MDA-
0.0

MB-231

Heart (fetal)
0.0
Breast ca.* (pl.ef) T47D
0.0

Heart
0.0
Breast ca. BT-549
1.6

Skeletal muscle (fetal)
0.0
Breast ca. MDA-N
0.0

Skeletal muscle
0.0
Ovary
0.0

Bone marrow
0.0
Ovarian ca. OVCAR-3
0.0

Thymus
0.0
Ovarian ca. OVCAR-4
0.0

Spleen
0.0
Ovarian ca. OVCAR-5
0.0

Lymph node
0.0
Ovarian ca. OVCAR-8
0.0

Colorectal
4.8
Ovarian ca. IGROV-1
0.0

Stomach
0.0
Ovarian ca.* (ascites)
4.2

SK-OV-3

Small intestine
0.0
Uterus
0.0

Colon ca. SW480
0.0
Plancenta
0.0

Colon ca.* SW620(SW480
0.0
Prostate
0.0

met)

Colon ca. HT29
0.0
Prostate ca.* (bone
0.0

met)PC-3

Colon ca. HCT-116
0.0
Testis
58.6

Colon ca. CaCo-2
0.0
Melanoma Hs688(A).T
0.0

Colon ca.
0.0
Melanoma* (met)
0.0

tissue(ODO3866)

Hs688(B).T

Colon ca. HCC-2998
0.0
Melanoma UACC-62
0.0

Gastric ca.* (liver met)
0.0
Melanoma M14
0.0

NCI-N87

Bladder
0.0
Melanoma LOX IMVI
0.0

Trachea
0.0
Melanoma* (met) SK-
0.0

MEL-5

Kidney
0.0
Adipose
0.0

[0750] Panel 1.3D Summary: Ag2625 The CG57376-01 gene is expressed at moderate levels exclusively in the testis and a renal cancer cell line in this panel (CTs=31.9-32.7). Thus, expression of this gene could be used to differentiate between those samples and other samples on this panel. Furthermore, the highly selective expression in the testis suggests that the protein encoded by this gene may be useful in the treatment of male infertility.

[0751] AK. CG59729-01: Olfactory Receptor

[0752] Expression of gene CG59729-01 was assessed using the primer-probe sets Ag2562, Ag1568 and Ag1569, described in Tables AKA, AKB and AKC. Results of the RTQ-PCR runs are shown in Tables AKD, AK-E, AKF and AKG.

160TABLE AKAProbe Name Ag2562StartSEQ IDPrimersSequencesLengthPositionNO:Forward5′-gtctttctgtgcctctcacatc-3′22503337ProbeTET-5′-ttttctgtgacacccagcctgtg-3′-TAMRA23535338Reverse5′-tgtcagagcaggagagctttag-3′22558339

[0753]

161

TABLE AKB

Probe Name Ag1568

Start
SEQ ID

Primers
Sequences
Length
Position
NO:

Forward
5′-gtctttctgtgcctctcacatc-3′
22
503
340

Probe
TET-5′-ttttctgtgacacccagcctgtg-3′-TAMRA
23
535
341

Reverse
5′-tgtcagagcaggagagctttag-3′
22
558
342

[0754]

162

TABLE AKC

Probe Name Ag1569

Start
SEQ ID

Primers
Sequences
Length
Position
NO:

Forward
5′-gtctttctgtgcctctcacatc-3′
22
503
343

Probe
TET-5′-ttttctgtgacacccagcctgtg-3′-TAMRA
23
535
344

Reverse
5′-tgtcagagcaggagagctttag-3′
22
558
345

[0755]

163

TABLE AKD

CNS_neurodegeneration_v1.0

Rel. Exp. (%) Ag2562, Run

Rel. Exp. (%) Ag2562, Run

Tissue Name
208779724
Tissue Name
208779724

AD 1 Hippo
12.8
Control (Path) 3
15.9

Temporal Ctx

AD 2 Hippo
35.1
Control (Path) 4
61.1

Temporal Ctx

AD 3 Hippo
4.7
AD 1 Occipital Ctx
13.4

AD 4 Hippo
36.1
AD 2 Occipital Ctx
0.0

(Missing)

AD 5 Hippo
36.1
AD 3 Occipital Ctx
1.7

AD 6 Hippo
37.9
AD 4 Occipital Ctx
42.6

Control 2 Hippo
17.1
AD 5 Occipital Ctx
36.1

Control 4 Hippo
19.5
AD 6 Occipital Ctx
23.5

Control (Path) 3 Hippo
19.5
Control 1 Occipital Ctx
4.4

AD 1 Temporal Ctx
23.0
Control 2 Occipital Ctx
20.6

AD 2 Temporal Ctx
38.7
Control 3 Occipital Ctx
37.4

AD 3 Temporal Ctx
3.3
Control 4 Occipital Ctx
25.0

AD 4 Temporal Ctx
61.6
Control (Path) 1
100.0

Occipital Ctx

AD 5 Inf Temporal Ctx
37.4
Control (Path) 2
18.6

Occipital Ctx

AD 5 Sup Temporal
34.2
Control (Path) 3
4.4

Ctx

Occipital Ctx

AD 6 Inf Temporal Ctx
50.0
Control (Path) 4
45.4

Occipital Ctx

AD 6 Sup Temporal
67.8
Control 1 Parietal Ctx
15.1

Ctx

Control 1 Temporal Ctx
14.1
Control 2 Parietal Ctx
48.3

Control 2 Temporal Ctx
13.1
Control 3 Parietal Ctx
18.4

Control 3 Temporal Ctx
18.9
Control (Path) 1
19.6

Parietal Ctx

Control 3 Temporal Ctx
34.9
Control (Path) 2
35.6

Parietal Ctx

Control (Path) 1
58.2
Control (Path) 3
20.6

Temporal Ctx

Parietal Ctx

Control (Path) 2
33.2
Control (Path) 4
81.8

Temporal Ctx

Parietal Ctx

[0756]

164

TABLE AKE

Panel 1.3D

Rel.
Rel.
Rel.

Rel.
Rel.
Rel.

Exp. (%)
Exp. (%)
Exp. (%)

Exp. (%)
Exp. (%)
Exp. (%)

Ag1568,
Ag1569,
Ag2562,

Ag1568,
Ag1569,
Ag2562,

Run
Run
Run

Run
Run
Run

Tissue Name
165534724
165529566
165672220
Tissue Name
165534724
165529566
165672220

Liver
0.0
8.7
7.7
Kidney
8.1
0.0
27.7

adenocarcinoma

(fetal)

Pancreas
12.6
0.0
0.0
Renal ca.
0.0
14.5
0.0

786-0

Pancreatic ca.
0.0
8.2
0.0
Renal ca.
34.2
24.0
11.8

CAPAN 2

A498

Adrenal gland
7.7
0.0
21.5
Renal ca.
17.3
43.8
9.1

RXF 393

Thyroid
13.1
0.0
0.0
Renal ca.
0.0
0.0
8.3

ACHN

Salivary gland
29.7
61.1
67.8
Renal ca.
0.0
0.0
0.0

UO-31

Pituitary gland
9.3
20.2
8.7
Renal ca.
0.0
0.0
14.7

TK-10

Brain (fetal)
7.1
0.0
14.3
Liver
22.1
26.4
0.0

Brain (whole)
0.0
16.4
0.0
Liver (fetal)
6.6
10.3
0.0

Brain (amygdala)
12.8
51.1
32.3
Liver ca.
10.3
0.0
0.0

(hepatoblast)

HepG2

Brain
27.9
7.2
19.6
Lung
7.3
10.6
47.3

(cerebellum)

Brain
24.7
16.3
45.1
Lung (fetal)
25.2
17.2
13.2

(hippocampus)

Brain (substantia
34.2
23.2
31.2
Lung ca.
0.0
0.0
0.0

nigra)

(small cell)

LX-1

Brain (thalamus)
100.0
39.8
22.1
Lung ca.
0.0
0.0
0.0

(small cell)

NCI-H69

Cerebral Cortex
6.3
43.8
11.2
Lung ca.
0.0
0.0
0.0

(s. cell var.)

SHP-77

Spinal cord
75.3
76.8
6.3
Lung ca.
0.0
33.0
18.8

(large

cell)NCI-

H460

glio/astro U87-
5.8
15.5
35.4
Lung ca.
9.3
14.8
0.0

MG

(non-sm. cell)

A549

glio/astro U-118-
0.0
0.0
21.0
Lung ca.
4.5
0.0
0.0

MG

(non-s. cell)

NCI-H23

astrocytoma
28.1
44.1
6.7
Lung ca.
0.0
9.3
4.7

SW1783

(non-s. cell)

HOP-62

neuro*; met SK-
6.0
4.0
6.5
Lung ca.
0.0
0.0
0.0

N-AS

(non-s. cl)

NCI-H522

astrocytoma SF-
9.3
15.0
0.0
Lung ca.
0.0
2.4
0.0

539

(squam.) SW

900

astrocytoma SNB-
10.7
10.4
4.7
Lung ca.
0.0
0.0
7.7

75

(squam.)

NCI-H596

glioma SNB-19
9.2
10.8
22.1
Mammary
0.0
0.0
11.1

gland

glioma U251
12.2
57.8
51.1
Breast ca.*
0.0
8.3
0.0

(pl. ef) MCF-7

glioma SF-295
36.6
7.7
26.6
Breast ca.*
0.0
9.3
7.6

(pl. ef) MDA-

MB-231

Heart (fetal)
0.0
0.0
0.0
Breast ca.*
0.0
20.7
0.0

(pl. ef) T47D

Heart
6.1
0.0
0.0
Breast ca.
0.0
0.0
0.0

BT-549

Skeletal muscle
6.0
0.0
0.0
Breast ca.
0.0
26.6
0.0

(fetal)

MDA-N

Skeletal muscle
0.0
0.0
15.3
Ovary
0.0
0.0
6.7

Bone marrow
0.0
32.1
6.3
Ovarian ca.
8.8
0.0
0.0

OVCAR-3

Thymus
36.1
9.6
0.0
Ovarian ca.
7.9
17.6
16.6

OVCAR-4

Spleen
18.4
29.9
0.0
Ovarian ca.
0.0
0.0
6.0

OVCAR-5

Lymph node
27.2
30.4
100.0
Ovarian ca.
3.4
0.0
15.3

OVCAR-8

Colorectal
77.9
31.2
39.0
Ovarian ca.
0.0
0.0
0.0

IGROV-1

Stomach
7.4
2.8
31.9
Ovarian ca.*
60.3
12.3
8.3

(ascites) SK-

OV-3

Small intestine
31.6
25.0
17.4
Uterus
8.4
100.0
45.4

Colon ca. SW480
7.6
6.7
7.5
Plancenta
12.2
51.4
24.7

Colon ca.*
0.0
0.0
0.0
Prostate
0.0
7.6
0.0

SW620(SW480

met)

Colon ca. HT29
0.0
0.0
0.0
Prostate ca.*
0.0
7.7
7.6

(bone

met)PC-3

Colon ca. HCT-
8.5
0.0
0.0
Testis
0.0
8.5
0.0

116

Colon ca. CaCo-2
15.4
0.0
0.0
Melanoma
0.0
0.0
7.5

Hs688(A).T

Colon ca.
0.0
0.0
0.0
Melanoma*
8.2
0.0
15.6

tissue(ODO3866)

(met)

Hs688(B).T

Colon ca. HCC-
28.9
0.0
0.0
Melanoma
0.0
0.0
12.4

2998

UACC-62

Gastric ca.* (liver
9.3
17.4
6.7
Melanoma
0.0
0.0
0.0

met) NCI-N87

M14

Bladder
62.4
62.0
17.8
Melanoma
0.0
0.0
0.0

LOX IMVI

Trachea
45.1
15.8
0.0
Melanoma*
9.6
0.0
0.0

(met) SK-

MEL-5

Kidney
0.0
7.9
0.0
Adipose
6.1
25.7
16.0

[0757]

165

TABLE AKF

Panel 2.2

Rel. Exp. (%)
Rel. Exp. (%)

Rel. Exp. (%)
Rel. Exp. (%)

Ag1568, Run
Ag1569, Run

Ag1568, Run
Ag1569, Run

Tissue Name
173968822
173850036
Tissue Name
173968822
173850036

Normal Colon
36.6
4.6
Kidney Margin
21.9
53.2

(OD04348)

Colon cancer
0.0
0.0
Kidney malignant
0.0
8.6

(OD06064)

cancer

(OD06204B)

Colon Margin
2.8
0.0
Kidney normal
6.7
0.0

(OD06064)

adjacent tissue

(OD06204E)

Colon cancer
0.0
0.0
Kidney Cancer
3.2
4.3

(OD06159)

(OD04450-01)

Colon Margin
35.4
12.4
Kidney Margin
23.0
27.0

(OD06159)

(OD04450-03)

Colon cancer
0.0
0.0
Kidney Cancer
0.0
0.0

(OD06297-04)

8120613

Colon Margin
2.8
19.1
Kidney Margin
15.2
2.9

(OD06297-015)

8120614

CC Gr.2 ascend
0.0
2.8
Kidney Cancer
7.2
0.0

colon (ODO3921)

9010320

CC Margin
4.9
4.2
Kidney Margin
11.7
0.0

(ODO3921)

9010321

Colon cancer
14.3
0.0
Kidney Cancer
0.0
0.0

metastasis

8120607

(OD06104)

Lung Margin
0.0
7.3
Kidney Margin
0.0
0.0

(OD06104)

8120608

Colon mets to lung
11.2
27.4
Normal Uterus
51.4
46.3

(OD04451-01)

Lung Margin
38.4
37.6
Uterine Cancer
23.0
28.1

(OD04451-02)

064011

Normal Prostate
21.6
13.1
Normal Thyroid
6.1
0.0

Prostate Cancer
22.7
23.7
Thyroid Cancer
0.0
0.0

(OD04410)

064010

Prostate Margin
15.7
5.0
Thyroid Cancer
27.7
26.1

(OD04410)

A302152

Normal Ovary
0.0
0.0
Thyroid Margin
0.0
19.2

A302153

Ovarian cancer
6.5
9.7
Normal Breast
66.4
15.0

(OD06283-03)

Ovarian Margin
25.3
30.1
Breast Cancer
22.5
15.8

(OD06283-07)

(OD04566)

Ovarian Cancer
12.0
10.3
Breast Cancer 1024
0.0
7.5

064008

Ovarian cancer
0.0
0.0
Breast Cancer
41.8
18.4

(OD06145)

(OD04590-01)

Ovarian Margin
21.8
7.5
Breast Cancer Mets
18.2
12.0

(OD06145)

(OD04590-03)

Ovarian cancer
0.0
0.0
Breast Cancer
37.1
17.6

(OD06455-03)

Metastasis

(OD04655-05)

Ovarian Margin
4.5
0.0
Breast Cancer
31.0
17.9

(OD06455-07)

064006

Normal Lung
12.0
5.9
Breast Cancer
19.2
3.2

9100266

Invasive poor diff.
6.5
3.9
Breast Margin
0.0
11.9

lung adeno

9100265

(ODO4945-01

Lung Margin
84.7
42.6
Breast Cancer
0.0
4.1

(ODO4945-03)

A209073

Lung Malignant
18.9
4.9
Breast Margin
44.4
29.5

Cancer (OD03126)

A2090734

Lung Margin
0.0
7.9
Breast cancer
42.9
37.9

(OD03126)

(OD06083)

Lung Cancer
0.0
31.2
Breast cancer node
30.6
37.4

(OD05014A)

metastasis

(OD06083)

Lung Margin
100.0
100.0
Normal Liver
20.3
13.2

(OD05014B)

Lung cancer
23.2
9.7
Liver Cancer 1026
0.0
4.5

(OD06081)

Lung Margin
45.4
7.3
Liver Cancer 1025
0.0
7.7

(OD06081)

Lung Cancer
5.7
0.0
Liver Cancer 6004-T
0.0
0.0

(OD04237-01)

Lung Margin
16.7
9.3
Liver Tissue 6004-N
0.0
3.9

(OD04237-02)

Ocular Melanoma
0.0
0.0
Liver Cancer 6005-T
0.0
0.0

Metastasis

Ocular Melanoma
0.0
4.3
Liver Tissue 6005-N
7.4
0.0

Margin (Liver)

Melanoma
0.0
0.0
Liver Cancer
10.7
0.0

Metastasis

064003

Melanoma Margin
11.1
10.7
Normal Bladder
12.8
16.0

(Lung)

Normal Kidney
22.1
4.0
Bladder Cancer
0.0
4.1

1023

Kidney Ca, Nuclear
19.9
24.1
Bladder Cancer
12.7
10.9

grade 2 (OD04338)

A302173

Kidney Margin
0.0
15.1
Normal Stomach
51.4
31.9

(OD04338)

Kidney Ca Nuclear
31.9
24.3
Gastric Cancer
0.0
0.0

grade 1/2

9060397

(OD04339)

Kidney Margin
10.3
0.7
Stomach Margin
11.3
8.7

(OD04339)

9060396

Kidney Ca, Clear
12.3
0.0
Gastric Cancer
19.8
0.0

cell type

9060395

(OD04340)

Kidney Margin
5.5
14.4
Stomach Margin
2.2
8.5

(OD04340)

9060394

Kidney Ca, Nuclear
0.0
3.6
Gastric Cancer
5.2
4.3

grade 3 (OD04348)

064005

[0758]

166

TABLE AKG

Panel 4D

Rel.
Rel.
Rel.

Rel.
Rel.
Rel.

Exp. (%)
Exp. (%)
Exp. (%)

Exp. (%)
Exp. (%)
Exp. (%)

Ag1568,
Ag1569,
Ag2562,

Ag1568,
Ag1569,
Ag2562,

Run
Run
Run

Run
Run
Run

Tissue Name
163479584
165301526
164393478
Tissue Name
163479584
165301526
164393478

Secondary Th1 act
13.0
9.5
31.0
HUVEC IL-
4.5
14.9
2.4

1beta

Secondary Th2 act
31.0
33.9
24.3
HUVEC IFN
69.7
60.7
65.1

gamma

Secondary Tr1 act
27.5
25.5
35.4
HUVEC TNF
9.6
9.2
14.7

alpha + IFN

gamma

Secondary Th1
39.8
12.2
22.7
HUVEC TNF
3.6
3.8
16.2

rest

alpha + IL4

Secondary Th2
94.6
59.9
27.2
HUVEC IL-11
57.8
46.3
35.6

rest

Secondary Tr1 rest
48.3
41.8
28.7
Lung
68.3
46.3
53.6

Microvascular

EC none

Primary Th1 act
6.1
0.0
5.5
Lung
33.0
51.8
41.8

Microvascular

EC TNF alpha +

IL-1beta

Primary Th2 act
19.2
11.2
3.5
Microvascular
55.5
77.4
55.9

Dermal EC none

Primary Tr1 act
10.8
2.3
14.2
Microsvasular
23.3
51.1
28.3

Dermal EC

TNF alpha + IL-

1beta

Primary Th1 rest
62.9
44.4
41.8
Bronchial
8.9
8.5
15.8

epithelium

TNF alpha +

IL1beta

Primary Th2 rest
47.6
26.8
57.0
Small airway
14.8
6.4
9.7

epithelium none

Primary Tr1 rest
14.6
13.9
8.1
Small airway
21.6
27.2
31.6

epithelium

TNF alpha + IL-

1beta

CD45RA CD4
16.3
5.8
2.7
Coronery artery
13.5
0.0
0.0

lymphocyte act

SMC rest

CD45RO CD4
19.5
18.2
25.5
Coronery artery
0.0
0.0
2.6

lymphocyte act

SMC TNF alpha +

IL-1beta

CD8 lymphocyte
15.6
9.3
21.0
Astrocytes rest
13.6
3.2
7.8

act

Secondary CD8
19.8
15.4
13.9
Astrocytes
3.4
2.6
2.2

lymphocyte rest

TNF alpha + IL-

1beta

Secondary CD8
21.0
11.8
21.0
KU-812
11.9
12.5
6.9

lymphocyte act

(Basophil) rest

CD4 lymphocyte
0.0
9.6
18.8
KU-812
29.1
24.8
25.5

none

(Basophil)

PMA/ionomycin

2ry
100.0
85.9
70.7
CCD1106
0.0
0.0
0.0

Th1/Th2/Tr1_anti-

(Keratinocytes)

CD95 CH11

none

LAK cells rest
28.3
23.0
10.7
CCD1106
0.0
5.1
0.0

(Keratinocytes)

TNF alpha + IL-

1beta

LAK cells IL-2
21.9
25.5
29.1
Liver cirrhosis
44.1
49.3
55.5

LAK cells IL-
30.8
11.3
26.6
Lupus kidney
0.0
0.0
9.0

2 + IL-12

LAK cells IL-
0.0
50.7
34.4
NCI-H292 none
26.2
33.4
14.8

2 + IFN gamma

LAK cells IL-2 +
44.8
39.8
37.6
NCI-H292 IL-4
12.3
2.4
19.2

IL-18

LAK cells
1.8
3.0
0.0
NCI-H292 IL-9
16.7
7.7
11.3

PMA/ionomycin

NK Cells IL-2 rest
14.6
33.4
26.2
NCI-H292 IL-13
5.8
1.6
6.8

Two Way MLR 3
32.5
18.9
25.0
NCI-H292 IFN
4.2
0.0
4.5

day

gamma

Two Way MLR 5
14.8
6.8
9.5
HPAEC none
58.2
35.4
71.2

day

Two Way MLR 7
12.2
3.4
22.2
HPAEC TNF
43.5
20.7
28.3

day

alpha + IL-1beta

PBMC rest
13.2
6.0
7.8
Lung fibroblast
10.2
7.0
27.7

none

PBMC PWM
33.2
48.6
39.5
Lung fibroblast
9.5
18.9
10.0

TNF alpha + IL-

1beta

PBMC PHA-L
15.4
11.7
11.9
Lung fibroblast
7.9
21.3
14.5

IL-4

Ramos (B cell)
0.0
7.2
2.9
Lung fibroblast
10.5
23.0
29.7

none

IL-9

Ramos (B cell)
12.8
7.1
24.5
Lung fibroblast
25.2
9.7
24.1

ionomycin

IL-13

B lymphocytes
48.6
29.3
24.1
Lung fibroblast
14.6
9.1
13.7

PWM

IFN gamma

B lymphocytes
40.6
47.3
38.7
Dermal
15.0
9.0
5.4

CD40L and IL-4

fibroblast

CCD1070 rest

EOL-1 dbcAMP
16.6
10.6
7.0
Dermal
57.8
43.2
48.0

fibroblast

CCD1070 TNF

alpha

EOL-1 dbcAMP
5.6
17.8
5.7
Dermal
7.1
2.3
10.7

PMA/ionomycin

fibroblast

CCD1070 IL-

1beta

Dendritic cells
10.4
12.7
14.7
Dermal
13.9
10.9
7.0

none

fibroblast IFN

gamma

Dendritic cells
14.4
15.1
13.6
Dermal
28.9
5.8
24.7

LPS

fibroblast IL-4

Dendritic cells
18.3
12.9
26.6
IBD Colitis 2
11.4
7.3
2.7

anti-CD40

Monocytes rest
2.5
1.3
13.9
IBD Crohn's
17.7
0.9
21.6

Monocytes LPS
11.0
8.5
11.5
Colon
29.1
17.7
15.0

Macrophages rest
9.2
14.9
12.2
Lung
8.1
12.2
10.6

Macrophages LPS
11.2
3.1
7.9
Thymus
33.2
25.3
27.2

HUVEC none
28.9
19.3
28.3
Kidney
94.0
100.0
100.0

HUVEC starved
84.7
65.1
93.3

[0759] CNS_neurodegeneration_v1.0 Summary: Ag2562 No difference is detected in the expression of the CG59729-01 gene in the postmortem brains of Alzheimer's diseased patients when compared to controls; however this panel demonstrates the expression of this gene in the brains of an independent group of subjects. Please see panel 1.3d for a discussion of utility of this gene in the central nervous system.

[0760] Panel 1.3D Summary: Ag1568/Ag1569/Ag2562 Three experiments with the same probe and primer set show expression of the CG59729-01 gene in samples from many different tissues. This gene represents a novel G-protein coupled receptor (GPCR) with expression in the brain. The GPCR family of receptors contains a large number of neurotransmitter receptors, including the dopamine, serotonin, a and b-adrenergic, acetylcholine muscarinic, histamine, peptide, and metabotropic glutamate receptors. GPCRs are excellent drug targets in various neurologic and psychiatric diseases. All antipsychotics have been shown to act at the dopamine D2 receptor; similarly novel antipsychotics also act at the serotonergic receptor, and often the muscarinic and adrenergic receptors as well. While the majority of antidepressants can be classified as selective serotonin reuptake inhibitors, blockade of the 5-HT1A and a2 adrenergic receptors increases the effects of these drugs. The GPCRs are also of use as drug targets in the treatment of stroke. Blockade of the glutamate receptors may decrease the neuronal death resulting from excitotoxicity; further more the purinergic receptors have also been implicated as drug targets in the treatment of cerebral ischemia. The b-adrenergic receptors have been implicated in the treatment of ADHD with Ritalin, while the a-adrenergic receptors have been implicated in memory. Therefore this gene may be of use as a small molecule target for the treatment of any of the described diseases.

[0761] There is also significant expression in a sample from uterus and a brain cancer cell line. Thus, expression of this gene could be used to differentiate between these and tissues and other samples on this panel.

[0762] Significant expression is also seen in the lymph node. Please see Panel 4D for discussion of potential utility in the immune system.

[0763] Overall, expression of this gene appears to be associated with normal tissue samples. This observation is in concordance with the results seen in Panel 2.2 (El Yacoubi et al., Adenosine A2A receptor antagonists are potential antidepressants: evidence based on pharmacology and A2A receptor knockout mice. Br J Pharmacol 134(1):68-77, 2001; Blier, Pharmacology of rapid-onset antidepressant treatment strategies. Clin Psychiatry 62 Suppl 15:12-7, 2001; Tranquillini and Reggiani, Glycine-site antagonists and stroke. Expert Opin Investig Drugs 8(11):1837-1848, 1999; Monopoli et al., Blockade of adenosine A2A receptors by SCH 58261 results in neuroprotective effects in cerebral ischaemia in rats. Neuroreport 9(17):3955-9, 1998).

[0764] Panel 2.2 Summary: Ag1568/Ag1569 Two experiments with the same probe and primer set show highest expression of the CG59729-01 gene in normal lung tissue adjacent to a tumor (CTs=32.5). Furthermore, there appears to be higher expression in a cluster of lung tissue samples when compared to matched tumor samples. There is also significant expression in normal kidney and uterus samples when compared to matched kidney and uterus cancers. Thus, expression of this gene could be used to differentiate between these samples and other samples in this panel. In addition, expression of this gene could also potentially be used as a marker to test for the presence of lung, kidney or uterine cancers. Therefore, therapeutic modulation of the expression or function of the protein encoded by this gene could potentially be useful in the treatment of lung, kidney and uterine cancers.

[0765] Panel 4D Summary: Ag1568/Ag1569/Ag2562 Three experiments with the same probe and primer set all show highest expression of the CG59729-01 transcript in kidney and secondary Th2 rest and secondary Th1/TH2/Tr1 cells treated with anti-CD95 (CTs=32.4). The expression of this transcript is decreased upon activation of these T (Th1 or Th2) cells. This transcript is also found at lower but still significant levels in B cells activated by PWM stimulation or treatment with CD40L and IL-4, where the latter condition promotes B cell survival and differentiation. This transcript encodes for a putative GPCR that may therefore function as regulator of Fas or other cell death pathways in T and B cells.

[0766] Expression of this transcript is also found in starved HUVEC, lung and dermal microvasculature. In addition, this expression of this transcript is down regulated in these tissues by TNF-a, a cytokine with cytotoxic activity on these cell types. Therefore, protein therapeutics designed with the putative GPCR encoded for by this protein could reduce or eliminate inflammation and tissue damage observed in lung and skin inflammatory diseases such as asthma, chronic bronchitis, psoriasis, and atopic dermatitis. Therapeutic modulation of the function or expression of the protein encoded by this gene may also reduce or eliminate inflammation and tissue damage that result from diseases associated with hyperactivated T cells including lupus erythematosus, rheumatoid arthritis, and inflammatory bowel diseases.

[0767] AL. CG59410-01: Olfactory Receptor

[0768] Expression of gene CG59410-01 was assessed using the primer-probe set Ag1709, described in Table ALA. Results of the RTQ-PCR runs are shown in Tables ALB and ALC.

167TABLE ALAProbe Name Ag1709StartSEQ IDPrimersSequencesLengthPositionNO:Forward5′-gctattctttcttgccaccttt-3′22599346ProbeTET-5′-tcagcacactactcatcgttctcaca-3′-TAMRA26628347Reverse5′-tggttacaacaatgaacgcata-3′22657348

[0769]

168

TABLE ALB

General_screening_panel_v1.5

Rel. Exp. (%) Ag1709, Run

Rel. Exp. (%) Ag1709, Run

Tissue Name
228980730
Tissue Name
228980730

Adipose
70.2
Renal ca. TK-10
1.2

Melanoma* Hs688(A).T
0.8
Bladder
2.6

Melanoma* Hs688(B).T
1.0
Gastric ca. (liver met.)
1.2

NCI-N87

Melanoma* M14
0.9
Gastric ca. KATO III
1.9

Melanoma* LOXIMVI
0.5
Colon ca. SW-948
2.0

Melanoma* SK-MEL-5
0.1
Colon ca. SW480
1.9

Squamous cell
0.7
Colon ca.* (SW480 met)
1.9

carcinoma SCC-4

SW620

Testis Pool
3.3
Colon ca. HT29
0.9

Prostate ca.* (bone met)
15.1
Colon ca. HCT-116
0.8

PC-3

Prostate Pool
1.6
Colon ca. CaCo-2
1.0

Placenta
2.2
Colon cancer tissue
0.9

Uterus Pool
0.9
Colon ca. SW1116
0.7

Ovarian ca. OVCAR-3
2.0
Colon ca. Colo-205
1.3

Ovarian ca. SK-OV-3
2.9
Colon ca. SW-48
1.0

Ovarian ca. OVCAR-4
1.8
Colon Pool
8.6

Ovarian ca. OVCAR-5
1.7
Small Intestine Pool
0.9

Ovarian ca. IGROV-1
4.2
Stomach Pool
100.0

Ovarian ca. OVCAR-8
1.4
Bone Marrow Pool
2.2

Ovary
1.9
Fetal Heart
1.0

Breast ca. MCF-7
2.0
Heart Pool
2.0

Breast ca. MDA-MB-
2.8
Lymph Node Pool
0.7

231

Breast ca. BT 549
2.1
Fetal Skeletal Muscle
1.7

Breast ca. T47D
1.4
Skeletal Muscle Pool
0.7

Breast ca. MDA-N
0.6
Spleen Pool
1.0

Breast Pool
1.6
Thymus Pool
1.5

Trachea
1.2
CNS cancer (glio/astro)
2.0

U87-MG

Lung
0.9
CNS cancer (glio/astro) U-
2.0

118-MG

Fetal Lung
1.3
CNS cancer (neuro;met)
0.8

SK-N-AS

Lung ca. NCI-N417
1.0
CNS cancer (astro) SF-539
2.3

Lung ca. LX-1
3.8
CNS cancer (astro) SNB-75
1.7

Lung ca. NCI-H146
0.5
CNS cancer (glio) SNB-19
1.1

Lung ca. SHP-77
0.5
CNS cancer (glio) SF-295
1.1

Lung ca. A549
0.5
Brain (Amygdala) Pool
1.7

Lung ca. NCI-H526
1.1
Brain (cerebellum)
1.8

Lung ca. NCI-H23
1.9
Brain (fetal)
2.1

Lung ca. NCI-H460
0.2
Brain (Hippocampus) Pool
1.4

Lung ca. HOP-62
3.2
Cerebral Cortex Pool
2.1

Lung ca. NCI-H522
4.1
Brain (Substantia nigra)
1.8

Pool

Liver
1.1
Brain (Thalamus) Pool
2.7

Fetal Liver
1.9
Brain (whole)
1.7

Liver ca. HepG2
0.6
Spinal Cord Pool
2.5

Kidney Pool
1.1
Adrenal Gland
1.7

Fetal Kidney
3.5
Pituitary gland Pool
2.8

Renal ca. 786-0
0.5
Salivary Gland
2.9

Renal ca. A498
1.9
Thyroid (female)
2.2

Renal ca. ACHN
25.7
Pancreatic ca. CAPAN2
1.4

Renal ca. UO-31
4.8
Pancreas Pool
1.4

[0770]

169

TABLE ALC

Panel 4D

Rel. Exp. (%) Ag1709,

Rel. Exp. (%) Ag1709,

Tissue Name
Run 165768294
Tissue Name
Run 165768294

Secondary Th1 act
0.0
HUVEC IL-1 beta
3.7

Secondary Th2 act
4.3
HUVEC IFN gamma
0.0

Secondary Tr1 act
9.7
HUVEC TNF alpha + IFN
3.3

gamma

Secondary Th1 rest
0.0
HUVEC TNF alpha + IL4
0.0

Secondary Th2 rest
0.0
HUVEC IL-11
0.0

Secondary Tr1 rest
0.0
Lung Microvascular EC none
0.0

Primary Th1 act
0.0
Lung Microvascular EC
0.0

TNF alpha + IL-1 beta

Primary Th2 act
0.0
Microvascular Dermal EC none
0.0

Primary Tr1 act
3.6
Microsvasular Dermal EC
0.0

TNF alpha + IL-1 beta

Primary Th1 rest
0.0
Bronchial epithelium TNF alpha +
0.0

IL1 beta

Primary Th2 rest
0.0
Small airway epithelium none
18.8

Primary Tr1 rest
1.9
Small airway epithelium
0.0

TNF alpha + IL-1 beta

CD45RA CD4 lymphocyte
0.0
Coronery artery SMC rest
0.0

act

CD45RO CD4 lymphocyte
10.7
Coronery artery SMC TNF alpha +
0.0

act

IL-1 beta

CD8 lymphocyte act
6.9
Astrocytes rest
0.0

Secondary CD8
13.6
Astrocytes TNF alpha + IL-1 beta
0.0

lymphocyte rest

Secondary CD8
0.0
KU-812 (Basophil) rest
4.9

lymphocyte act

CD4 lymphocyte none
11.1
KU-812 (Basophil)
5.1

PMA/ionomycin

2ry Th1/Th2/Tr1_anti-
0.0
CCD1106 (Keratinocytes) none
0.0

CD95 CH11

LAK cells rest
0.0
CCD1106 (Keratinocytes)
0.0

TNF alpha + IL-1 beta

LAK cells IL-2
0.0
Liver cirrhosis
100.0

LAK cells IL-2 + IL-12
0.0
Lupus kidney
0.0

LAK cells IL-2 + IFN
0.0
NCI-H292 none
0.0

gamma

LAK cells IL-2 + IL-18
0.0
NCI-H292 IL-4
0.0

LAK cells
8.9
NCI-H292 IL-9
0.0

PMA/ionomycin

NK Cells IL-2 rest
0.0
NCI-H292 IL-13
6.4

Two Way MLR 3 day
8.7
NCI-H292 IFN gamma
4.5

Two Way MLR 5 day
5.7
HPAEC none
0.0

Two Way MLR 7 day
0.0
HPAEC TNF alpha + IL-1 beta
0.0

PBMC rest
9.7
Lung fibroblast none
8.2

PBMC PWM
0.0
Lung fibroblast TNF alpha + IL-
0.0

1 beta

PBMC PHA-L
7.5
Lung fibroblast IL-4
6.1

Ramos (B cell) none
3.6
Lung fibroblast IL-9
0.0

Ramos (B cell) ionomycin
0.0
Lung fibroblast IL-13
0.0

B lymphocytes PWM
3.7
Lung fibroblast IFN gamma
0.0

B lymphocytes CD40L
0.0
Dermal fibroblast CCD1070 rest
0.0

and IL-4

EOL-1 dbcAMP
0.0
Dermal fibroblast CCD1070
18.4

TNF alpha

EOL-1 dbcAMP
0.0
Dermal fibroblast CCD1070 IL-
10.0

PMA/ionomycin

1 beta

Dendritic cells none
5.9
Dermal fibroblast IFN gamma
0.0

Dendritic cells LPS
3.7
Dermal fibroblast IL-4
0.0

Dendritic cells anti-CD40
0.0
IBD Colitis 2
58.6

Monocytes rest
0.0
IBD Crohn's
13.6

Monocytes LPS
0.0
Colon
11.2

Macrophages rest
0.0
Lung
7.0

Macrophages LPS
0.0
Thymus
0.0

HUVEC none
0.0
Kidney
0.0

HUVEC starved
0.0

[0771] General_screening_panel_v1.5 Summary: Ag1709 The expression of the CG59410-01 gene appears to be highest in a sample derived from normal stomach (CT=27.5). In addition, there is substantial expression in samples derived from normal colon tissue and adipose tissue, as well as a renal cell cancer cell line (ACHN). In the majority of samples, there was rather low level, uniform expression. Thus, the expression of this gene could be used to distinguish the above listed samples from the other samples in this panel.

[0772] This gene represents a novel G-protein coupled receptor (GPCR) and also shows expression in the brain. The GPCR family of receptors contains a large number of neurotransmitter receptors, including the dopamine, serotonin, a and b-adrenergic, acetylcholine muscarinic, histamine, peptide, and metabotropic glutamate receptors. GPCRs are excellent drug targets in various neurologic and psychiatric diseases. All antipsychotics have been shown to act at the dopamine D2 receptor; similarly novel antipsychotics also act at the serotonergic receptor, and often the muscarinic and adrenergic receptors as well. While the majority of antidepressants can be classified as selective serotonin reuptake inhibitors, blockade of the 5-HT1A and a2 adrenergic receptors increases the effects of these drugs. The GPCRs are also of use as drug targets in the treatment of stroke. Blockade of the glutamate receptors may decrease the neuronal death resulting from excitotoxicity; further more the purinergic receptors have also been implicated as drug targets in the treatment of cerebral ischemia. The b-adrenergic receptors have been implicated in the treatment of ADHD with Ritalin, while the a-adrenergic receptors have been implicated in memory. Therefore this gene may be of use as a small molecule target for the treatment of any of the described diseases (El Yacoubi et al., Adenosine A2A receptor antagonists are potential antidepressants: evidence based on pharmacology and A2A receptor knockout mice. Br J Pharmacol 134(1):68-77, 2001; Blier, Pharmacology of rapid-onset antidepressant treatment strategies. Clin Psychiatry 62 Suppl 15:12-7, 2001; Tranquillini and Reggiani, Glycine-site antagonists and stroke. Expert Opin Investig Drugs 8(11):1837-1848, 1999; Monopoli et al., Blockade of adenosine A2A receptors by SCH 58261 results in neuroprotective effects in cerebral ischaemia in rats. Neuroreport 9(17):3955-9, 1998).

[0773] Panel 4D Summary: Ag1709 Expression of the CG59410-01 gene is restricted to liver cirrhosis and IBD Colitis (CTs=33-34). The function of the putative GPCR encoded by the CG59410-01 gene may thus be important in the disease processes in both inflammatory bowel disease and in liver cirrhosis. Therefore, blocking antibodies or small molecule antagonists targeted to this GPCR may be useful as therapeutics in colitis and in cirrhosis.

[0774] AM. CG59400-01: Olfactory Receptor

[0775] Expression of gene CG59400-01 was assessed using the primer-probe sets Ag1581 and Ag 1727, described in Tables AMA and AMB. Results of the RTQ-PCR runs are shown in Table AMC.

170TABLE AMAProbe Name Ag1581StartSEQ IDPrimersSequencesLengthPositionNO:Forward5′-aattcattttgctgggactga-3′2135349ProbeTET-5′-ttcttctctttgccctcttctcggtt-3′-TAMRA2677350Reverse5′-acccaaaactgtgaccacatag-3′22105351

[0776]

171

TABLE AMB

Probe Name Ag1727

Start
SEQ ID

Primers
Sequences
Length
Position
NO:

Forward
5′-aattcattttgctgggactga-3′
21
35
352

Probe
TET-5′-ttcttctctttgccctcttctcggtt-3′-TAMRA
26
77
353

Reverse
5′-acccaaaactgtgaccacatag-3′
22
105
354

[0777]

172

TABLE AMC

Panel 4D

Rel. Exp. (%)
Rel. Exp. (%)

Rel. Exp. (%)
Rel. Exp. (%)

Ag1581, Run
Ag1727, Run

Ag1581, Run
Ag1727, Run

Tissue Name
165373544
165767201
Tissue Name
165373544
165767201

Secondary Th1 act
0.0
0.0
HUVEC IL-1beta
0.0
0.0

Secondary Th2 act
0.0
0.0
HUVEC IFN gamma
0.0
0.0

Secondary Tr1 act
0.0
0.0
HUVEC TNF alpha +
0.0
0.0

IFN gamma

Secondary Th1 rest
0.0
0.0
HUVEC TNF alpha +
0.0
0.0

IL4

Secondary Th2 rest
0.0
0.0
HUVEC IL-11
0.0
0.0

Secondary Tr1 rest
0.0
0.0
Lung Microvascular
0.0
0.0

EC none

Primary Th1 act
0.0
0.0
Lung Microvascular
0.0
0.0

EC TNF alpha + IL-

1beta

Primary Th2 act
94.0
0.0
Microvascular
0.0
0.0

Dermal EC none

Primary Tr1 act
100.0
7.2
Microsvasular Dermal
68.8
0.0

EC TNF alpha + IL-

1beta

Primary Th1 rest
0.0
0.0
Bronchial epithelium
0.0
0.0

TNF alpha + IL1beta

Primary Th2 rest
0.0
0.0
Small airway
0.0
0.0

epithelium none

Primary Tr1 rest
0.0
0.0
Small airway
0.0
0.0

epithelium TNF alpha +

IL-1beta

CD45RA CD4
0.0
0.0
Coronery artery SMC
0.0
0.0

lymphocyte act

rest

CD45RO CD4
75.8
0.0
Coronery artery SMC
0.0
0.0

lymphocyte act

TNF alpha + IL-1beta

CD8 lymphocyte act
0.0
0.0
Astrocytes rest
0.0
0.0

Secondary CD8
0.0
0.0
Astrocytes TNF alpha +
0.0
0.0

lymphocyte rest

IL-1beta

Secondary CD8
0.0
0.0
KU-812 (Basophil)
0.0
0.0

lymphocyte act

rest

CD4 lymphocyte
0.0
0.0
KU-812 (Basophil)
0.0
0.0

none

PMA/ionomycin

2ry
0.0
0.0
CCD1106
0.0
0.0

Th1/Th2/Tr1_anti-

(Keratinocytes) none

CD95 CH11

LAK cells rest
0.0
0.0
CCD1106
0.0
0.0

(Keratinocytes)

TNF alpha + IL-1beta

LAK cells IL-2
0.0
0.0
Liver cirrhosis
57.0
100.0

LAK cells IL-2 + IL-
0.0
0.0
Lupus kidney
0.0
0.0

12

LAK cells IL-2 + IFN
0.0
0.0
NCI-H292 none
0.0
0.0

gamma

LAK cells IL-2 + IL-
0.0
0.0
NCI-H292 IL-4
0.0
0.0

18

LAK cells
0.0
0.0
NCI-H292 IL-9
0.0
0.0

PMA/ionomycin

NK Cells IL-2 rest
0.0
0.0
NCI-H292 IL-13
0.0
0.0

Two Way MLR 3
0.0
0.0
NCI-H292 IFN
0.0
0.0

day

gamma

Two Way MLR 5
0.0
0.0
HPAEC none
0.0
0.0

day

Two Way MLR 7
0.0
0.0
HPAEC TNF alpha +
0.0
0.7

day

IL-1beta

PBMC rest
0.0
0.0
Lung fibroblast none
0.0
0.0

PBMC PWM
0.0
0.0
Lung fibroblast TNF
0.0
0.0

alpha + IL-1beta

PBMC PHA-L
0.0
0.0
Lung fibroblast IL-4
0.0
0.0

Ramos (B cell) none
0.0
0.0
Lung fibroblast IL-9
0.0
0.0

Ramos (B cell)
0.0
0.0
Lung fibroblast IL-13
0.0
0.0

ionomycin

B lymphocytes PWM
0.0
0.0
Lung fibroblast IFN
0.0
1.1

gamma

B lymphocytes
0.0
5.7
Dermal fibroblast
0.0
0.0

CD40L and IL-4

CCD1070 rest

EOL-1 dbcAMP
0.0
0.0
Dermal fibroblast
0.0
0.0

CCD1070 TNF alpha

EOL-1 dbcAMP
0.0
0.0
Dermal fibroblast
0.0
0.0

PMA/ionomycin

CCD1070 IL-1beta

Dendritic cells none
0.0
0.0
Dermal fibroblast IFN
0.0
0.0

gamma

Dendritic cells LPS
0.0
1.0
Dermal fibroblast IL-4
0.0
0.0

Dendritic cells anti-
74.2
0.0
IBD Colitis 2
85.3
16.2

CD40

Monocytes rest
0.0
0.0
IBD Crohn's
0.0
0.0

Monocytes LPS
0.0
0.0
Colon
0.0
8.8

Macrophages rest
0.0
0.0
Lung
0.0
0.0

Macrophages LPS
0.0
0.0
Thymus
0.0
0.0

HUVEC none
0.0
0.0
Kidney
0.0
0.0

HUVEC starved
78.5
0.0

[0778] Panel 1.3D Summary: Ag1581 The CG59400-01 gene shows low/undetectable levels of expression in all samples on this panel (CTs>35). (Data not shown.)

[0779] Panel 2.2 Summary: Ag1581 The CG59400-01 gene shows low/undetectable levels of expression in all samples on this panel (CTs>35). (Data not shown.)

[0780] Panel 4D Summary: Ag1727 The CG59400-01 transcript is only detected in liver cirrhosis. Furthermore, this transcript is not detected in normal liver in Panel 1.3D, suggesting that CG59400-01 gene expression is unique to liver cirrhosis. The CG59400-01 gene encodes a putative GPCR; therefore, antibodies or small molecule therapeutics could reduce or inhibit fibrosis that occurs in liver cirrhosis. In addition, antibodies to this putative GPCR could also be used for the diagnosis of liver cirrhosis. One run with Ag1581 had low/undetectable levels of expression (CT>35) in all samples in the panel. (Data not shown.)

[0781] AN. CG110254-01: Olfactory Receptor

[0782] Expression of gene CG110254-01 was assessed using the primer-probe sets Ag2696 and Ag1790, described in Tables ANA and ANB. Results of the RTQ-PCR runs are shown in Tables ANC, AND, and ANE.

173TABLE ANAProbe Name Ag2696StartSEQ IDPrimersSequencesLengthPositionNO:Forward5′-actacgtgccacctgtctgtat-3′22772355ProbeTET-5′-ctacctgcagcctcgctccagtgag-3′-TAMRA25794356Reverse5′-agcattggagttacgattgtgt-3′22847357

[0783]

174

TABLE ANB

Probe Name Ag1790

Start
SEQ ID

Primers
Sequences
Length
Position
NO:

Forward
5′-cctgtacacagtcatggcctat-3′
22
359
358

Probe
TET-5′-atctgtcaacccctgcactacccagt-3′-TAMRA
26
396
359

Reverse
5′-atttctgcacacatccttctgt-3′
22
430
360

[0784]

175

TABLE ANC

Panel 1.3D

Rel. Exp. (%) Ag1790, Run

Rel. Exp. (%) Ag1790, Run

Tissue Name
165974809
Tissue Name
165974809

Liver adenocarcinoma
0.0
Kidney (fetal)
0.0

Pancreas
8.7
Renal ca. 786-0
0.0

Pancreatic ca. CAPAN 2
0.0
Renal ca. A498
0.0

Adrenal gland
0.0
Renal ca. RXF 393
54.0

Thyroid
0.0
Renal ca. ACHN
0.0

Salivary gland
0.0
Renal ca. UO-31
0.0

Pituitary gland
0.0
Renal ca. TK-10
0.0

Brain (fetal)
0.0
Liver
0.0

Brain (whole)
3.7
Liver (fetal)
0.0

Brain (amygdala)
10.6
Liver ca. (hepatoblast)
0.0

HepG2

Brain (cerebellum)
6.8
Lung
0.0

Brain (hippocampus)
0.0
Lung (fetal)
0.0

Brain (substantia nigra)
0.0
Lung ca. (small cell) LX-1
0.0

Brain (thalamus)
0.0
Lung ca. (small cell)
0.0

NCI-H69

Cerebral Cortex
0.0
Lung ca. (s.cell var.)
0.0

SHP-77

Spinal cord
0.0
Lung ca. (large cell)NCI-
0.0

H460

glio/astro U87-MG
0.0
Lung ca. (non-sm.cell)
0.0

A549

glio/astro U-118-MG
0.0
Lung ca. (non-s.cell)
0.0

NCI-H23

astrocytoma SW1783
0.0
Lung ca. (non-s.cell)
0.0

HOP-62

neuro*; met SK-N-AS
0.0
Lung ca. (non-s.cell) NCI-
0.0

H522

astrocytoma SF-539
0.0
Lung ca. (squam.) SW
0.0

900

astrocytoma SNB-75
0.0
Lung ca. (squam.) NCI-
0.0

H596

glioma SNB-19
0.0
Mammary gland
0.0

glioma U251
0.0
Breast ca.* (pl.ef) MCF-7
0.0

glioma SF-295
0.0
Breast ca.* (pl.ef) MDA-
0.0

MB-231

Heart (fetal)
0.0
Breast ca.* (pl.ef) T47D
0.0

Heart
0.0
Breast ca. BT-549
0.0

Skeletal muscle (fetal)
0.0
Breast ca. MDA-N
0.0

Skeletal muscle
0.0
Ovary
0.0

Bone marrow
0.0
Ovarian ca. OVCAR-3
0.0

Thymus
0.0
Ovarian ca. OVCAR-4
0.0

Spleen
100.0
Ovarian ca. OVCAR-5
0.0

Lymph node
0.0
Ovarian ca. OVCAR-8
0.0

Colorectal
20.0
Ovarian ca. IGROV-1
0.0

Stomach
0.0
Ovarian ca.* (ascites)
0.0

SK-OV-3

Small intestine
0.0
Uterus
0.0

Colon ca. SW480
0.0
Plancenta
0.0

Colon ca.* SW620(SW480
0.0
Prostate
0.0

met)

Colon ca. HT29
0.0
Prostate ca.* (bone
7.6

met)PC-3

Colon ca. HCT-116
0.0
Testis
0.0

Colon ca. CaCo-2
0.0
Melanoma Hs688(A).T
0.0

Colon ca.
0.0
Melanoma* (met)
0.0

tissue(ODO3866)

Hs688(B).T

Colon ca. HCC-2998
0.0
Melanoma UACC-62
0.0

Gastric ca.* (liver met)
0.0
Melanoma M14
0.0

NCI-N87

Bladder
6.1
Melanoma LOX IMVI
0.0

Trachea
0.0
Melanoma* (met) SK-
0.0

MEL-5

Kidney
0.0
Adipose
0.0

[0785]

176

TABLE AND

Panel 2D

Rel. Exp. (%) Ag2696,

Rel. Exp. (%) Ag2696,

Tissue Name
Run 153291452
Tissue Name
Run 153291452

Normal Colon
21.6
Kidney Margin 8120608
0.0

CC Well to Mod Diff
10.2
Kidney Cancer 8120613
0.0

(ODO3866)

CC Margin (ODO3866)
19.2
Kidney Margin 8120614
0.0

CC Gr.2 rectosigmoid
0.0
Kidney Cancer 9010320
0.0

(ODO3868)

CC Margin (ODO3868)
0.0
Kidney Margin 9010321
0.0

CC Mod Diff (ODO3920)
12.9
Normal Uterus
0.0

CC Margin (ODO3920)
10.4
Uterus Cancer 064011
0.0

CC Gr.2 ascend colon
0.0
Normal Thyroid
0.0

(ODO3921)

CC Margin (ODO3921)
0.0
Thyroid Cancer 064010
0.0

CC from Partial Hepatectomy
0.0
Thyroid Cancer A302152
0.0

(ODO4309) Mets

Liver Margin (ODO4309)
0.0
Thyroid Margin A302153
0.0

Colon mets to lung (OD04451-
0.0
Normal Breast
0.0

01)

Lung Margin (OD04451-02)
0.0
Breast Cancer (OD04566)
0.0

Normal Prostate 6546-1
0.0
Breast Cancer (OD04590-
0.0

01)

Prostate Cancer (OD04410)
0.0
Breast Cancer Mets
0.0

(OD04590-03)

Prostate Margin (OD04410)
0.0
Breast Cancer Metastasis
0.0

(OD04655-05)

Prostate Cancer (OD04720-01)
0.0
Breast Cancer 064006
0.0

Prostate Margin (OD04720-02
0.0
Breast Cancer 1024
6.9

Normal Lung 061010
14.2
Breast Cancer 9100266
0.0

Lung Met to Muscle
0.0
Breast Margin 9100265
0.0

(ODO4286)

Muscle Margin (OD04286)
0.0
Breast Cancer A209073
0.0

Lung Malignant Cancer
0.0
Breast Margin A2090734
0.0

(OD03126)

Lung Margin (OD03126)
0.0
Normal Liver
0.0

Lung Cancer (OD04404)
100.0
Liver Cancer 064003
0.0

Lung Margin (OD04404)
0.0
Liver Cancer 1025
0.0

Lung Cancer (OD04565)
0.0
Liver Cancer 1026
0.0

Lung Margin (0D04565)
0.0
Liver Cancer 6004-T
0.0

Lung Cancer (OD04237-01)
0.0
Liver Tissue 6004-N
0.0

Lung Margin (OD04237-02)
0.0
Liver Cancer 6005-T
0.0

Ocular Mel Met to Liver
0.0
Liver Tissue 6005-N
0.0

(ODO4310)

Liver Margin (ODO4310)
0.0
Normal Bladder
43.2

Melanoma Mets to Lung
0.0
Bladder Cancer 1023
0.0

(OD04321)

Lung Margin (OD04321)
0.0
Bladder Cancer A302173
0.0

Normal Kidney
8.8
Bladder Cancer
0.0

(OD04718-01)

Kidney Ca, Nuclear grade 2
0.0
Bladder Normal Adjacent
13.2

(OD04338)

(OD04718-03)

Kidney Margin (OD04338)
0.0
Normal Ovary
0.0

Kidney Ca Nuclear grade 1/2
0.0
Ovarian Cancer 064008
0.0

(OD04339)

Kidney Margin (OD04339)
0.0
Ovarian Cancer
0.0

(OD04768-07)

Kidney Ca, Clear cell type
0.0
Ovary Margin (OD04768-
0.0

(OD04340)

08)

Kidney Margin (OD04340)
14.2
Normal Stomach
0.0

Kidney Ca, Nuclear grade 3
0.0
Gastric Cancer 9060358
0.0

(OD04348)

Kidney Margin (OD04348)
0.0
Stomach Margin 9060359
11.3

Kidney Cancer (OD04622-01)
0.0
Gastric Cancer 9060395
0.0

Kidney Margin (OD04622-03)
0.0
Stomach Margin 9060394
0.0

Kidney Cancer (OD04450-01)
0.0
Gastric Cancer 9060397
0.0

Kidney Margin (OD04450-03)
2.3
Stomach Margin 9060396
12.4

Kidney Cancer 8120607
0.0
Gastric Cancer 064005
0.0

[0786]

177

TABLE ANE

Panel 4D

Rel. Exp. (%) Ag1790,

Rel. Exp. (%) Ag1790,

Tissue Name
Run 165801864
Tissue Name
Run 165801864

Secondary Th1 act
0.0
HUVEC IL-1 beta
0.0

Secondary Th2 act
0.0
HUVEC IFN gamma
0.0

Secondary Tr1 act
0.0
HUVEC TNF alpha + IFN
0.0

gamma

Secondary Th1 rest
1.8
HUVEC TNF alpha + IL4
7.5

Secondary Th2 rest
0.0
HUVEC IL-11
0.0

Secondary Tr1 rest
0.0
Lung Microvascular EC none
0.0

Primary Th1 act
0.0
Lung Microvascular EC
0.0

TNF alpha + IL-1 beta

Primary Th2 act
0.0
Microvascular Dermal EC none
0.0

Primary Tr1 act
0.0
Microsvasular Dermal EC
0.0

TNF alpha + IL-1 beta

Primary Th1 rest
0.0
Bronchial epithelium TNF alpha +
5.7

IL1 beta

Primary Th2 rest
0.0
Small airway epithelium none
3.9

Primary Tr1 rest
0.0
Small airway epithelium
56.3

TNF alpha + IL-1 beta

CD45RA CD4 lymphocyte
0.0
Coronery artery SMC rest
0.0

act

CD45RO CD4 lymphocyte
0.0
Coronery artery SMC TNF alpha +
10.0

act

IL-1 beta

CD8 lymphocyte act
0.0
Astrocytes rest
4.0

Secondary CD8
0.0
Astrocytes TNF alpha + IL-1 beta
0.0

lymphocyte rest

Secondary CD8
0.0
KU-812 (Basophil) rest
0.0

lymphocyte act

CD4 lymphocyte none
0.0
KU-812 (Basophil)
0.0

PMA/ionomycin

2ry Th1/Th2/Tr1_anti-
0.0
CCD1106 (Keratinocytes) none
3.8

CD95 CH11

LAK cells rest
0.0
CCD1106 (Keratinocytes)
36.6

TNF alpha + IL-1 beta

LAK cells IL-2
0.0
Liver cirrhosis
100.0

LAK cells IL-2 + IL-12
0.0
Lupus kidney
8.8

LAK cells IL-2 + IFN
0.0
NCI-H292 none
0.0

gamma

LAK cells IL-2 + IL-18
0.0
NCI-H292 IL-4
0.0

LAK cells
0.0
NCI-H292 IL-9
0.0

PMA/ionomycin

NK Cells IL-2 rest
0.0
NCI-H292 IL-13
0.0

Two Way MLR 3 day
0.0
NCI-H292 IFN gamma
0.0

Two Way MLR 5 day
0.0
HPAEC none
2.6

Two Way MLR 7 day
0.0
HPAEC TNF alpha + IL-1 beta
65.1

PBMC rest
0.0
Lung fibroblast none
0.0

PBMC PWM
0.0
Lung fibroblast TNF alpha + IL-
0.0

1 beta

PBMC PHA-L
0.0
Lung fibroblast IL-4
0.0

Ramos (B cell) none
0.0
Lung fibroblast IL-9
0.0

Ramos (B cell) ionomycin
0.0
Lung fibroblast IL-13
0.0

B lymphocytes PWM
5.0
Lung fibroblast IFN gamma
0.0

B lymphocytes CD40L
3.7
Dermal fibroblast CCD1070 rest
0.0

and IL-4

EOL-1 dbcAMP
0.0
Dermal fibroblast CCD1070
3.4

TNF alpha

EOL- 1 dbcAMP
0.0
Dermal fibroblast CCD1070 IL-
0.0

PMA/ionomycin

1 beta

Dendritic cells none
0.0
Dermal fibroblast IFN gamma
0.0

Dendritic cells LPS
0.0
Dermal fibroblast IL-4
0.0

Dendritic cells anti-CD40
0.0
IBD Colitis 2
5.7

Monocytes rest
0.0
IBD Crohn's
0.0

Monocytes LPS
0.0
Colon
6.0

Macrophages rest
0.0
Lung
8.1

Macrophages LPS
0.0
Thymus
20.7

HUVEC none
0.0
Kidney
8.5

HUVEC starved
0.0

[0787] CNS_neurodegeneration_v1.0 Summary: Ag2696 Expression of the CG110254-01 gene is low/undetectable (CT>35) in all samples in this panel. (Data not shown.)

[0788] Panel 1.3D Summary: Ag1790 The CG110254-01 gene is only expressed in the spleen (CT=33.9), an important site of secondary immune responses. Therefore, antibodies or small molecule therapeutics that block the function of this GPCR may be useful as anti-inflammatory therapeutics for the treatment of allergies, autoimmune diseases, and inflammatory diseases. Ag2696 Expression of the CG110254-01 gene is low/undetectable (CT>35) in all samples in this panel. (Data not shown.)

[0789] Panel 2.2 Summary: Ag1790 Expression of the CG110254-01 gene is low/undetectable (CT>35) in all samples in this panel. (Data not shown.)

[0790] Panel 2D Summary: Ag2696 Expression of the CG110254-01 gene is restricted to a lung cancer (CT=34.3). This gene appears to be overexpressed in lung cancer when compared to adjacent normal tissue. Therefore, expression of this gene could be used to as a marker for lung cancer. Furthermore, therapeutic modulation of the expression or function of the protein product may be effective in the treatment of lung cancer.

[0791] Panel 4D Summary: Ag1790 The CG110254-01 gene is expressed at highest levels in small airway epithelium (CT=33.5). Therefore, expression of this gene could be used to distinguish small airway epithelium from the other samples on this panel. Furthermore, antibodies or small molecule drugs that inhibit the action of the CG110254-01 gene product may reduce or eliminate the symptoms in patients with asthma, allergies, and chronic obstructive pulmonary disease. Ag2696 Expression of this gene is low/undetectable (CT>35) in all of the samples in this panel (data not shown).

[0792] AO. CG50303-03/GMAC073079_B: Olfactory Receptor

[0793] Expression of gene CG50303-03 was assessed using the primer-probe sets Ag2377, Ag2607, Ag2610, Ag1501 and Ag1585, described in Tables AOA, AOB, AOC, AOD and AOE. Results of the RTQ-PCR runs are shown in Tables AOF, AOG, AOH, AOI, AOJ, AOK and AOL.

178TABLE AOAProbe Name Ag2377StartSEQ IDPrimersSequencesLengthPositionNO:Forward5′-atgggaaacaccatcatcatag-3′22130361ProbeTET-5′-tggtcatagctgacacccacctacat-3′-TAMRA26155362Reverse5′-aattgcccaggaagaagtacat-3′22187363

[0794]

179

TABLE AOB

Probe Name Ag2607

Start
SEQ ID

Primers
Sequences
Length
Position
NO:

Forward
5′-catagctgacacccacctacat-3′
22
130
364

Probe
TET-5′-cacccatgtacttcttcctgggcaat-3′-TAMRA
26
182
365

Reverse
5′-actgcagtcatggttaccaaga-3′
22
224
366

[0795]

180

TABLE AOC

Probe Name Ag2610

Start
SEQ ID

Primers
Sequences
Length
Position
NO:

Forward
5′-gtctcacctcacactggtcttc-3′
22
738
367

Probe
TET-5′-catctttctgtatgtcaggcctggca-3′-TAMRA
26
777
368

Reverse
5′-ctgacttgcacagagtgagctt-3′
22
803
369

[0796]

181

TABLE AOD

Probe Name Ag1501

Start
SEQ ID

Primers
Sequences
Length
Position
NO:

Forward
5′-catagctgacacccacctacat-3′
22
159
370

Probe
TET-5′-cacccatgtacttcttcctgggcaat-3′-TAMRA
26
182
371

Reverse
5′-ctgcagtcatggttaccaagat-3′
22
223
372

[0797]

182

TABLE AOE

Probe Name Ag1585

Start
SEQ ID

Primers
Sequences
Length
Position
NO:

Forward
5′-catagctgacacccacctacat-3′
22
159
373

Probe
TET-5′-cacccatgtacttcttcctgggcaat-3′-TAMRA
26
182
374

Reverse
5′-ctgcagtcatggttaccaagat-3′
22
223
375

[0798]

183

TABLE AOF

CNS_neurodegeneration_v1.0

Rel.
Rel.
Rel.

Rel.
Rel.
Rel.

Exp. (%)
Exp. (%)
Exp. (%)

Exp. (%)
Exp. (%)
Exp. (%)

Ag2377,
Ag2607,
Ag2610,

Ag2377,
Ag2607,
Ag2610,

Run
Run
Run
Tissue
Run
Run
Run

Tissue Name
208271229
208971580
208393679
Name
208271229
208971580
208393679

AD 1 Hippo
9.2
4.5
14.9
Control
1.4
1.4
0.0

(Path) 3

Temporal

Ctx

AD 2 Hippo
10.7
15.1
45.7
Control
18.2
35.1
36.9

(Path) 4

Temporal

Ctx

AD 3 Hippo
8.8
8.5
18.7
AD 1
16.3
35.6
17.7

Occipital

Ctx

AD 4 Hippo
7.2
9.0
4.8
AD 2
0.0
0.0
0.0

Occipital

Ctx

(Missing)

AD 5 hippo
32.5
44.4
48.0
AD 3
10.3
13.0
18.9

Occipital

Ctx

AD 6 Hippo
100.0
35.6
33.9
AD 4
16.4
57.4
20.0

Occipital

Ctx

Control 2
24.0
33.4
22.5
AD 5
11.8
27.5
22.4

Hippo

Occipital

Ctx

Control 4
5.6
7.4
13.9
AD 6
3.6
18.0
13.6

Hippo

Occipital

Ctx

Control (Path)
4.0
6.2
0.0
Control 1
7.2
7.7
2.6

3 Hippo

Occipital

Ctx

AD 1
36.6
60.3
27.4
Control 2
0.1
55.5
25.3

Temporal Ctx

Occipital

Ctx

AD 2
18.3
39.5
46.3
Control 3
19.3
29.1
32.3

Temporal Ctx

Occipital

Ctx

AD 3
10.1
10.3
18.9
Control 4
10.0
20.9
8.2

Temporal Ctx

Occipital

Ctx

AD 4
30.4
52.5
31.9
Control
62.9
100.0
100.0

Temporal Ctx

(Path) 1

Occipital

Ctx

AD 5 Inf
35.1
85.3
52.5
Control
23.5
47.6
12.9

Temporal Ctx

(Path) 2

Occipital

Ctx

AD 5
10.7
36.6
28.7
Control
1.1
2.2
5.2

SupTemporal

(Path) 3

Ctx

Occipital

Ctx

AD 6 Inf
27.5
87.7
60.7
Control
30.8
46.0
39.2

Temporal Ctx

(Path) 4

Occipital

Ctx

AD 6 Sup
22.4
72.2
52.5
Control 1
14.3
35.1
10.0

Temporal Ctx

Parietal

Ctx

Control 1
3.9
15.6
6.0
Control 2
17.4
61.1
30.6

Temporal Ctx

Parietal

Ctx

Control 2
7.6
11.6
13.0
Control 3
15.5
26.2
25.7

Temporal Ctx

Parietal

Ctx

Control 3
9.7
12.6
4.4
Control
27.2
60.3
64.6

Temporal Ctx

(Path) 1

Parietal

Ctx

Control 4
11.5
15.4
6.8
Control
57.0
54.7
43.8

Temporal Ctx

(Path) 2

Parietal

Ctx

Control (Path)
43.8
67.4
36.3
Control
2.0
0.0
7.3

1 Temporal

(Path) 3

Ctx

Parietal

Ctx

Control (Path)
17.8
35.6
22.4
Control
48.3
65.5
82.9

2 Temporal

(Path) 4

Ctx

Parietal

Ctx

[0799]

184

TABLE AOG

Panel 1.2

Rel. Exp. (%) Ag1501, Run

Rel. Exp. (%) Ag1501, Run

Tissue Name
140466099
Tissue Name
140466099

Endothelial cells
6.8
Renal ca. 786-0
14.3

Heart (Fetal)
0.3
Renal ca. A498
12.6

Pancreas
0.6
Renal ca. RXF 393
15.9

Pancreatic ca. CAPAN 2
0.3
Renal ca. ACHN
11.8

Adrenal Gland
16.4
Renal ca. UO-31
21.0

Thyroid
0.0
Renal ca. TK-10
28.3

Salivary gland
38.2
Liver
7.6

Pituitary gland
1.2
Liver (fetal)
5.1

Brain (fetal)
20.6
Liver ca. (hepatoblast)
9.9

HepG2

Brain (whole)
13.3
Lung
0.0

Brain (amygdala)
17.2
Lung (fetal)
0.9

Brain (cerebellum)
7.0
Lung ca. (small cell) LX-1
33.9

Brain (hippocampus)
67.4
Lung ca. (small cell)
58.6

NCI-H69

Brain (thalamus)
92.0
Lung ca. (s.cell var.)
3.6

SHP-77

Cerebral Cortex
85.3
Lung ca. (large cell)NCI-
24.0

H460

Spinal cord
25.0
Lung ca. (non-sm.cell)
30.8

A549

glio/astro U87-MG
17.0
Lung ca. (non-s.cell)
81.8

NCI-H23

glio/astro U-118-MG
5.5
Lung ca. (non-s.cell)
24.3

HOP-62

astrocytoma SW1783
7.2
Lung ca. (non-s.cl) NCI-
59.0

H522

neuro*; met SK-N-AS
1.5
Lung ca. (squam.) SW
10.4

900

astrocytoma SF-539
2.4
Lung ca. (squam.) NCI-
25.7

H596

astrocytoma SNB-75
14.2
Mammary gland
15.3

glioma SNB-19
23.5
Breast ca.* (pl.ef) MCF-7
2.1

glioma U251
6.9
Breast ca.* (pl.ef) MDA-
2.3

MB-231

glioma SF-295
18.8
Breast ca.* (pl. ef) T47D
88.9

Heart
50.7
Breast ca. BT-549
6.1

Skeletal Muscle
13.2
Breast ca. MDA-N
74.7

Bone marrow
7.5
Ovary
0.7

Thymus
0.0
Ovarian ca. OVCAR-3
14.3

Spleen
4.6
Ovarian ca. OVCAR-4
40.3

Lymph node
2.9
Ovarian ca. OVCAR-5
72.7

Colorectal Tissue
9.3
Ovarian ca. OVCAR-8
100.0

Stomach
0.7
Ovarian ca. IGROV-1
56.3

Small intestine
6.5
Ovarian ca. (ascites) SK-
16.4

OV-3

Colon ca. SW480
1.5
Uterus
9.5

Colon ca.* SW620
16.2
Placenta
39.0

(SW480 met)

Colon ca. HT29
14.7
Prostate
8.8

Colon ca. HCT-116
7.6
Prostate ca.* (bone met)
39.0

PC-3

Colon ca. CaCo-2
9.4
Testis
14.0

Colon ca. Tissue
31.2
Melanoma Hs688(A).T
1.3

(ODO3866)

Colon ca. HCC-2998
10.3
Melanoma* (met)
10.6

Hs688(B).T

Gastric ca.* (liver met)
21.3
Melanoma UACC-62
48.6

NCI-N87

Bladder
36.6
Melanoma M14
69.7

Trachea
0.0
Melanoma LOX IMVI
0.0

Kidney
35.8
Melanoma* (met) SK-
9.7

MEL-5

Kidney (fetal)
14.9

[0800]

185

TABLE AOH

Panel 1.3D

Rel. Exp. (%)
Rel. Exp. (%)
Rel. Exp. (%)
Rel. Exp. (%)

Ag1585, Run
Ag2377, Run
Ag2607, Run
Ag2610, Run

Tissue Name
165529870
165631765
166219825
166162989

Liver adenocarcinoma
5.1
5.6
0.0
0.0

Pancreas
0.0
0.0
0.0
0.0

Pancreatic ca. CAPAN 2
0.0
0.0
0.0
5.6

Adrenal gland
0.0
0.0
0.0
0.0

Thyroid
0.0
0.0
0.0
0.0

Salivary gland
11.0
4.5
7.0
0.0

Pituitary gland
0.0
0.0
7.7
28.9

Brain (fetal)
31.2
53.2
12.7
21.0

Brain (whole)
56.3
48.6
100.0
100.0

Brain (amygdala)
15.4
0.0
31.9
0.0

Brain (cerebellum)
10.0
19.6
0.0
25.2

Brain (hippocampus)
5.5
20.3
17.3
7.0

Brain (substantia
54.3
80.7
49.3
31.6

nigra)

Brain (thalamus)
26.2
58.2
55.5
72.2

Cerebral Cortex
5.6
0.0
14.3
0.0

Spinal cord
100.0
100.0
98.6
38.7

glio/astro U87-MG
13.0
0.0
0.0
0.0

glio/astro U-118-MG
4.2
7.2
0.0
0.0

astrocytoma SW1783
0.0
0.0
19.9
0.0

neuro*; met SK-N-AS
0.0
0.0
0.0
0.0

astrocytoma SF-539
12.3
6.5
0.0
5.4

astrocytoma SNB-75
0.0
0.0
0.0
3.9

glioma SNB-19
2.1
4.0
4.5
18.2

glioma U251
19.3
0.0
0.0
0.0

glioma SF-295
23.5
0.0
0.0
0.0

Heart (fetal)
0.0
0.0
0.0
0.0

Heart
12.4
10.7
0.0
0.0

Skeletal muscle (fetal)
0.0
0.0
0.0
0.0

Skeletal muscle
6.1
0.0
0.0
0.0

Bone marrow
0.0
0.0
0.0
0.0

Thymus
0.0
0.0
0.0
0.0

Spleen
0.0
0.0
0.0
0.0

Lymph node
6.7
0.0
0.0
0.0

Colorectal
10.7
4.9
0.0
0.0

Stomach
0.0
0.0
0.0
7.2

Small intestine
0.0
0.0
0.0
0.0

Colon ca. SW480
0.0
0.0
0.0
5.2

Colon ca.*
6.8
0.0
6.8
8.5

SW620(SW480 met)

Colon ca. HT29
0.0
0.0
0.0
0.0

Colon ca. HCT-116
5.9
0.0
0.0
0.0

Colon ca. CaCo-2
0.0
6.6
0.0
0.0

Colon ca.
0.0
0.0
0.0
0.0

tissue(ODO3866)

Colon ca. HCC-2998
0.0
0.0
0.0
0.0

Gastric ca.* (liver met)
0.0
6.7
0.0
8.0

NCI-N87

Bladder
2.8
0.0
9.9
17.4

Trachea
0.0
6.4
0.0
0.0

Kidney
0.0
0.0
0.0
0.0

Kidney (fetal)
0.0
0.0
0.0
12.3

Renal ca. 786-0
0.0
3.4
0.0
0.0

Renal ca. A498
6.9
0.0
0.0
6.1

Renal ca. RXF 393
0.0
9.5
0.0
7.9

Renal ca. ACHN
0.0
0.0
3.1
6.4

Renal ca. UO-31
11.0
0.0
0.0
0.0

Renal ca. TK-10
4.9
0.0
0.0
0.0

Liver
8.0
0.0
0.0
0.0

Liver (fetal)
0.0
0.0
0.0
0.0

Liver ca. (hepatoblast)
0.0
0.0
0.0
0.0

HepG2

Lung
11.0
0.0
0.0
0.0

Lung (fetal)
0.0
0.0
0.0
0.0

Lung ca. (small cell)
3.5
14.0
7.4
0.0

LX-1

Lung ca. (small cell)
0.0
0.0
0.0
0.0

NCI-H69

Lung ca. (s.cell var.)
0.0
0.0
0.0
4.1

SHP-77

Lung ca. (large
3.3
0.0
0.0
0.0

cell)NCI-H460

Lung ca. (non-sm.cell)
0.0
0.0
0.0
0.0

A549

Lung ca. (non-s.cell)
5.3
14.4
8.2
5.6

NCI-H23

Lung ca. (non-s.cell)
0.0
0.0
0.0
0.0

HOP-62

Lung ca. (non-s.cl)
0.0
0.0
0.0
0.0

NCI-H522

Lung ca. (squam.) SW
7.2
5.4
0.0
0.0

900

Lung ca. (squam.)
0.0
0.0
0.0
0.0

NCI-H596

Mammary gland
0.0
0.0
7.7
0.0

Breast ca.* (pl.ef)
4.7
0.0
0.0
0.0

MCF-7

Breast ca.* (pl.ef)
0.0
0.0
0.0
6.0

MDA-MB-231

Breast ca.* (pl.ef)
14.0
0.0
6.9
29.3

T47D

Breast ca. BT-549
0.0
0.0
0.0
0.0

Breast ca. MDA-N
5.2
12.6
18.0
0.0

Ovary
0.0
0.0
0.0
0.0

Ovarian ca. OVCAR-3
0.0
0.0
15.8
0.0

Ovarian ca. OVCAR-4
0.0
0.0
0.0
5.0

Ovarian ca. OVCAR-5
7.7
0.0
11.7
4.5

Ovarian ca. OVCAR-8
29.7
15.9
18.3
8.0

Ovarian ca. IGROV-1
17.6
0.0
0.0
0.0

Ovarian ca.* (ascites)
0.0
0.0
0.0
6.4

SK-OV-3

Uterus
0.0
9.5
6.1
0.0

Plancenta
51.8
21.5
43.2
39.0

Prostate
0.0
14.6
0.0
0.0

Prostate ca.* (bone
0.0
0.0
6.0
0.0

met)PC-3

Testis
17.4
14.8
18.6
10.5

Melanoma Hs688(A).T
0.0
0.0
0.0
0.0

Melanoma* (met)
0.0
0.0
0.0
0.0

Hs688(B).T

Melanoma UACC-62
0.0
3.7
0.0
6.9

Melanoma M14
6.4
35.6
12.1
0.0

Melanoma LOX IMVI
0.0
0.0
0.0
0.0

Melanoma* (met) SK-
0.0
0.0
0.0
10.4

MEL-5

Adipose
0.0
0.0
0.0
0.0

[0801]

186

TABLE AOI

Panel 2.2

Rel. Exp. (%)
Rel. Exp. (%)

Rel. Exp. (%)
Rel. Exp. (%)

Ag2377, Run
Ag2607, Run

Ag2377, Run
Ag2607, Run

Tissue Name
174553776
175128152
Tissue Name
174553776
175128152

Normal Colon
0.0
0.0
Kidney Margin
0.0
20.7

(OD04348)

Colon cancer
15.3
0.0
Kidney malignant
24.1
21.8

(OD06064)

cancer

(OD06204B)

Colon Margin
0.0
0.0
Kidney normal
0.0
0.0

(OD06064)

adjacent tissue

(OD06204E)

Colon cancer
0.0
0.0
Kidney Cancer
0.0
9.5

(OD06159)

(OD04450-01)

Colon Margin
0.0
0.0
Kidney Margin
0.0
0.0

(OD06159)

(OD04450-03)

Colon cancer
0.0
0.0
Kidney Cancer
0.0
0.0

(OD06297-04)

8120613

Colon Margin
0.0
0.0
Kidney Margin
0.0
0.0

(OD06297-015)

8120614

CC Gr.2 ascend
0.0
0.0
Kidney Cancer
0.0
0.0

colon (ODO3921)

9010320

CC Margin
0.0
0.0
Kidney Margin
0.0
0.0

(ODO3921)

9010321

Colon cancer
0.0
0.0
Kidney Cancer
0.0
0.0

metastasis

8120607

(OD06104)

Lung Margin
0.0
0.0
Kidney Margin
0.0
0.0

(OD06104)

8120608

Colon mets to lung
27.9
0.0
Normal Uterus
35.6
17.8

(OD04451-01)

Lung Margin
32.5
0.0
Uterine Cancer
0.0
0.0

(OD04451-02)

064011

Normal Prostate
0.0
0.0
Normal Thyroid
0.0
0.0

Prostate Cancer
0.0
0.0
Thyroid Cancer
0.0
0.0

(OD04410)

064010

Prostate Margin
0.0
0.0
Thyroid Cancer
0.0
0.0

(OD04410)

A302152

Normal Ovary
0.0
0.0
Thyroid Margin
0.0
0.0

A302153

Ovarian cancer
10.7
0.0
Normal Breast
15.0
8.7

(OD06283-03)

Ovarian Margin
0.0
0.0
Breast Cancer
0.0
0.0

(OD06238-07)

(OD04566)

Ovarian Cancer
0.0
24.1
Breast Cancer 1024
84.1
36.1

064008

Ovarian cancer
0.0
0.0
Breast Cancer
0.0
0.0

(OD06145)

(OD04590-01)

Ovarian Margin
34.9
0.0
Breast Cancer Mets
22.5
0.0

(OD06145)

(OD04590-03)

Ovarian cancer
24.7
0.0
Breast Cancer
0.0
0.0

(OD06455-03)

Metastasis

(OD04655-05)

Ovarian Margin
9.9
0.0
Breast Cancer
0.0
19.1

(OD06455-07)

064006

Normal Lung
0.0
9.2
Breast Cancer
100.0
100.0

9100266

Invasive poor diff.
12.7
0.0
Breast Margin
9.9
0.0

lung adeno

9100265

(ODO4945-01

Lung Margin
0.0
18.3
Breast Cancer
0.0
0.0

(ODO4945-03)

A209073

Lung Malignant
0.0
0.0
Breast Margin
14.6
0.0

Cancer (OD03126)

A2090734

Lung Margin
0.0
0.0
Breast cancer
75.8
9.9

(OD03126)

(OD06083)

Lung Cancer
0.0
0.0
Breast cancer node
16.6
25.3

(OD05014A)

metastasis

(OD06083)

Lung Margin
19.5
6.0
Normal Liver
0.0
0.0

(OD05014B)

Lung cancer
25.9
9.3
Liver Cancer 1026
0.0
0.0

(OD06081)

Lung Margin
0.0
0.0
Liver Cancer 1025
33.4
6.7

(OD06081)

Lung Cancer
0.0
0.0
Liver Cancer 6004-T
0.0
0.0

(OD04237-01)

Lung Margin
0.0
11.0
Liver Tissue 6004-N
0.0
0.0

(OD04237-02)

Ocular Melanoma
13.6
0.0
Liver Cancer 6005-T
0.0
0.0

Metastasis

Ocular Melanoma
0.0
0.0
Liver Tissue 6005-N
0.0
9.7

Margin (Liver)

Melanoma
0.0
2.6
Liver Cancer
0.0
0.0

Metastasis

064003

Melanoma Margin
0.0
0.0
Normal Bladder
0.0
0.0

(Lung)

Normal Kidney
17.9
0.0
Bladder Cancer
0.0
0.0

1023

Kidney Ca, Nuclear
0.0
0.0
Bladder Cancer
0.0
0.0

grade 2 (OD04338)

A302173

Kidney Margin
0.0
0.0
Normal Stomach
0.0
0.0

(OD04338)

Kidney Ca Nuclear
15.3
0.0
Gastric Cancer
0.0
0.0

grade 1/2

9060397

(OD04339)

Kidney Margin
0.0
0.0
Stomach Margin
0.0
7.6

(OD04339)

9060396

Kidney Ca, Clear
0.0
0.0
Gastric Cancer
33.2
7.6

cell type

9060395

(OD04340)

Kidney Margin
0.0
12.2
Stomach Margin
0.0
0.0

(OD04340)

9060394

Kidney Ca, Nuclear
0.0
0.0
Gastric Cancer
0.0
0.0

grade 3 (OD04348)

064005

[0802]

187

TABLE AOJ

Panel 4D

Rel. Exp. (%)
Rel. Exp. (%)

Rel. Exp. (%)
Rel. Exp. (%)

Ag2377, Run
Ag2607, Run

Ag2377, Run
Ag2607, Run

Tissue Name
164216614
164160833
Tissue Name
164216614
164160833

Secondary Th1 act
27.4
14.3
HUVEC IL-1beta
0.0
0.0

Secondary Th2 act
36.3
0.0
HUVEC IFN gamma
0.0
4.0

Secondary Tr1 act
10.5
9.4
HUVEC TNF alpha +
0.0
4.0

IFN gamma

Secondary Th1 rest
0.0
0.0
HUVEC TNF alpha +
12.4
6.7

IL4

Secondary Th2 rest
0.0
0.0
HUVEC IL-11
12.6
0.0

Secondary Tr1 rest
12.9
0.0
Lung Microvascular
18.7
16.7

EC none

Primary Th1 act
0.0
0.0
Lung Microvascular
0.0
12.9

EC TNF alpha + IL-

1beta

Primary Th2 act
8.1
0.0
Microvascular
35.1
4.5

Dermal EC none

Primary Tr1 act
25.3
10.3
Microsvasular Dermal
8.2
10.7

EC TNF alpha + IL-

1beta

Primary Th1 rest
28.1
53.2
Bronchial epithelium
0.0
0.0

TNF alpha + IL1beta

Primary Th2 rest
69.3
23.3
Small airway
0.0
0.0

epithelium none

Primary Tr1 rest
13.3
15.1
Small airway
0.0
10.7

epithelium TNF alpha

+ IL-1beta

CD45RA CD4
0.0
0.0
Coronery artery SMC
12.2
0.0

lymphocyte act

rest

CD45RO CD4
34.4
7.3
Coronery artery SMC
0.0
0.0

lymphocyte act

TNF alpha + IL-1beta

CD8 lymphocyte act
8.7
9.2
Astrocytes rest
19.1
5.1

Secondary CD8
31.0
13.1
Astrocytes TNF alpha
8.9
0.0

lymphocyte rest

+ IL-1beta

Secondary CD8
12.2
10.8
KU-812 (Basophil)
0.0
0.0

lymphocyte act

rest

CD4 lymphocyte
0.0
0.0
KU-812 (Basophil)
13.9
5.6

none

PMA/ionomycin

2ry
25.9
14.6
CCD1106
10.7
4.3

Th1/Th2/Tr1_anti-

(Keratinocytes) none

CD95 CH11

LAK cells rest
28.5
33.2
CCD1106
12.7
0.0

(Keratinocytes)

TNF alpha + IL-1beta

LAK cells IL-2
0.0
9.9
Liver cirrhosis
81.8
50.0

LAK cells IL-2 + IL-
20.0
0.0
Lupus kidney
23.2
8.6

12

LAK cells IL-2 + IFN
12.2
10.0
NCI-H292 none
0.0
3.7

gamma

LAK cells IL-2 + IL-
9.4
0.0
NCI-H292 IL-4
44.4
0.0

18

LAK cells
18.9
0.0
NCI-H292 IL-9
7.0
0.0

PMA/ionomycin

NK Cells IL-2 rest
25.5
17.4
NCI-H292 IL-13
0.0
3.9

Two Way MLR 3
31.2
5.4
NCI-H292 IFN
11.0
1.9

day

gamma

Two Way MLR 5
0.0
12.0
HPAEC none
6.3
11.3

day

Two Way MLR 7
10.6
0.0
HPAEC TNF alpha +
24.3
4.5

day

IL-1beta

PBMC rest
11.4
5.0
Lung fibroblast none
0.0
0.0

PBMC PWM
12.2
19.5
Lung fibroblast TNF
0.0
0.0

alpha + IL-1beta

PBMC PHA-L
28.1
18.6
Lung fibroblast IL-4
0.0
0.0

Ramos (B cell) none
13.9
18.8
Lung fibroblast IL-9
0.0
3.3

Ramos (B cell)
16.6
7.8
Lung fibroblast IL-13
0.0
0.0

ionomycin

B lymphocytes PWM
15.7
12.9
Lung fibroblast IFN
0.0
5.1

gamma

B lymphocytes
24.8
0.0
Dermal fibroblast
2.4
15.7

CD40L and IL-4

CCD1070 rest

EOL-1 dbcAMP
13.7
7.4
Dermal fibroblast
66.4
10.4

CCD1070 TNF alpha

EOL-1 dbcAMP
6.2
0.0
Dermal fibroblast
0.0
0.0

PMA/ionomycin

CCD1070 IL-1beta

Dendritic cells none
15.5
29.5
Dermal fibroblast IFN
15.1
7.5

gamma

Dendritic cells LPS
10.7
17.0
Dermal fibroblast IL-4
0.0
0.0

Dendritic cells anti-
11.5
5.6
IBD Colitis 2
10.7
0.0

CD40

Monocytes rest
0.0
0.0
IBD Crohn's
0.0
0.0

Monocytes LPS
100.0
50.3
Colon
0.0
0.0

Macrophages rest
76.8
100.0
Lung
0.0
4.5

Macrophages LPS
8.0
8.5
Thymus
24.1
13.7

HUVEC none
9.0
0.0
Kidney
44.4
17.7

HUVEC starved
41.8
2.8

[0803]

188

TABLE AOK

Panel CNS_1

Rel. Exp. (%)
Rel. Exp. (%)

Rel. Exp. (%)
Rel. Exp. (%)

Ag2377, Run
Ag2377, Run

Ag2377, Run
Ag2377, Run

Tissue Name
171656285
182012511
Tissue Name
171656285
182012511

BA4 Control
6.1
7.4
BA17 PSP
0.0
21.0

BA4 Control2
8.8
4.2
BA17 PSP2
11.1
10.4

BA4
0.0
7.1
Sub Nigra
42.3
41.2

Alzheimer's2

Control

BA4
24.7
19.8
Sub Nigra
29.5
3.6

Parkinson's

Control2

BA4
17.8
30.8
Sub Nigra
28.5
12.6

Parkinson's2

Alzheimer's2

BA4
17.6
3.7
Sub Nigra
55.1
61.1

Huntington's

Parkinson's2

BA4
8.1
0.0
Sub Nigra
100.0
100.0

Huntington's2

Huntington's

BA4 PSP
38.2
12.2
Sub Nigra
17.3
21.2

Huntington's2

BA4 PSP2
20.0
5.2
Sub Nigra PSP2
9.7
5.4

BA4
49.7
31.2
Sub Nigra
87.1
42.0

Depression

Depression

BA4
14.2
18.0
Sub Nigra
33.0
20.4

Depression2

Depression2

BA7 Control
23.3
2.7
Glob Palladus
28.5
25.7

Control

BA7 Control2
25.5
11.5
Glob Palladus
25.2
15.2

Control2

BA7
18.9
4.4
Glob Palladus
11.9
16.4

Alzheimer's2

Alzheimer's

BA7
11.4
9.8
Glob Palladus
4.2
36.9

Parkinson's

Alzheimer's2

BA7
0.0
14.6
Glob Palladus
37.9
44.4

Parkinson's

Parkinson's

BA7
23.7
10.9
Glob Palladus
9.0
26.1

Huntington's

Parkinson's2

BA7
42.9
26.8
Glob Palladus
48.0
33.4

Huntington's2

PSP

BA7 PSP
30.8
14.6
Glob Palladus
10.7
9.9

PSP2

BA7 PSP2
4.2
10.4
Glob Palladus
40.9
39.5

Depression

BA7
31.9
21.3
Temp Pole
0.0
0.0

Depression

Control

BA9 Control
2.0
4.4
Temp Pole
11.8
7.9

Control2

BA9 Control2
16.7
24.7
Temp Pole
0.0
0.0

Alzheimer's

BA9
0.0
6.6
Temp Pole
0.0
3.4

Alzheimer's

Alzheimer's2

BA9
2.9
0.0
Temp Pole
17.3
7.1

Alzheimer's2

Parkinson's

BA9
11.3
15.2
Temp Pole
0.0
9.5

Parkinson's

Parkinson's2

BA9
7.9
4.2
Temp Pole
0.0
4.9

Parkinson's

Huntington's

BA9
39.8
14.5
Temp Pole PSP
6.7
6.3

Huntington's

BA9
8.1
3.7
Temp Pole PSP2
0.0
0.0

Huntington's2

BA9 PSP
44.4
5.7
Temp Pole
0.0
23.8

Depression2

BA9 PSP2
0.0
0.0
Cing Gyr Control
31.2
27.4

BA9
15.1
5.9
Cing Gyr
16.8
24.5

Depression

Control2

BA9
14.4
8.7
Cing Gyr
17.8
13.2

Depression2

Alzheimer's

BA17 Control
47.0
30.4
Cing Gyr
13.9
3.4

Alzheimer's2

BA17 Control2
28.7
5.4
Cing Gyr
26.2
30.8

Parkinson's

BA17
7.5
7.1
Cing Gyr
24.8
25.9

Alzheimer's2

Parkinson's2

BA17
38.2
68.3
Cing Gyr
30.8
28.7

Parkinson's

Huntington's

BA17
24.0
9.3
Cing Gyr
20.7
14.2

Parkinson's2

Huntington's2

BA17
36.1
13.8
Cing Gyr PSP
90.1
76.3

Huntington's

BA17
15.2
16.4
Cing Gyr PSP2
0.0
20.3

Huntington's2

BA17
58.6
27.7
Cing Gyr
52.5
61.1

Depression

Depression

BA17
65.5
60.3
Cing Gyr
43.5
15.3

Depression2

Depression2

[0804]

189

TABLE AOL

Panel CNS_1.1

Rel. Exp. (%)
Rel. Exp. (%)

Rel. Exp. (%)
Rel. Exp. (%)

Ag2377, Run
Ag2377, Run

Ag2377, Run
Ag2377, Run

Tissue Name
200060897
200061715
Tissue Name
200060897
200061715

Cing Gyr
39.2
13.9
BA17 PSP2
5.3
11.8

Depression2

Cing Gyr
35.8
23.2
BA17 PSP
4.2
13.1

Depression

Cing Gyr PSP2
6.2
2.8
BA17
17.8
10.1

Huntington's2

Cing Gyr PSP
100.0
100.0
BA17
36.9
6.9

Huntington's

Cing Gyr
32.5
10.4
BA17
19.2
12.2

Huntington's2

Parkinson's2

Cing Gyr
27.4
8.8
BA17
37.4
19.1

Huntington's

Parkinson's

Cing Gyr
12.8
1.9
BA17
7.7
0.0

Parkinson's2

Alzheimer's2

Cing Gyr
47.6
32.5
BA17 Control2
35.8
20.0

Parkinson's

Cing Gyr
0.0
7.2
BA17 Control
35.1
22.7

Alzheimer's2

Cing Gyr
13.8
3.6
BA9
8.6
3.8

Alzheimer's

Depression2

Cing Gyr
77.9
1.4
BA9
0.0
14.1

Control2

Depression

Cing Gyr Control
30.1
11.7
BA9 PSP2
3.6
12.4

Temp Pole
0.0
14.8
BA9 PSP
48.6
18.3

Depression2

Temp Pole PSP2
0.0
4.5
BA9
6.9
5.0

Huntington's2

Temp Pole PSP
5.5
3.3
BA9
59.0
8.4

Huntington's

Temp Pole
0.0
7.7
BA9
0.0
2.5

Huntington's

Parkinson's2

Temp Pole
0.0
0.0
BA9
7.9
0.0

Parkinson's2

Parkinson's

Temp Pole
27.5
4.9
BA9
0.0
0.0

Parkinson's

Alzheimer's2

Temp Pole
0.0
0.0
BA9
0.0
0.0

Alzheimer's2

Alzheimer's

Temp Pole
0.0
0.0
BA9 Control2
26.4
12.2

Alzheimer's

Temp Pole
21.3
8.4
BA9 Control
15.1
0.0

Control2

Temp Pole
0.0
0.0
BA7
29.3
11.1

Control

Depression

Glob Palladus
35.8
16.4
BA7 PSP2
28.7
2.9

Depression

Glob Palladus
5.5
5.0
BA7 PSP
7.0
6.6

PSP2

Glob Palladus
23.7
8.6
BA7
18.6
23.7

PSP

Huntington's2

Glob Palladus
34.2
6.5
BA7
11.3
6.7

Parkinson's2

Huntington's

Glob Palladus
16.4
20.3
BA7
0.0
0.0

Parkinson's

Parkinson's2

Glob Palladus
19.5
3.4
BA7
9.5
1.2

Alzheimer's2

Parkinson's

Glob Palladus
24.3
6.7
BA7
19.6
0.0

Alzheimer's

Alzheimer's2

Glob Palladus
13.8
2.8
BA7 Control2
25.3
2.4

Control2

Glob Palladus
33.2
17.7
BA7 Control
10.1
9.6

Control

Sub Nigra
36.3
5.5
BA4
27.5
15.9

Depression2

Depression2

Sub Nigra
52.5
10.4
BA4
10.8
15.6

Depression

Depression

Sub Nigra PSP2
12.4
15.9
BA4 PSP2
15.2
17.0

Sub Nigra
8.7
5.9
BA4 PSP
11.3
10.7

Huntington's2

Sub Nigra
82.4
51.1
BA4
0.0
0.0

Huntington's

Huntington's2

Sub Nigra
34.9
12.5
BA4
0.0
3.8

Parkinson's2

Huntington's

Sub Nigra
34.2
15.0
BA4
18.7
11.7

Alzheimer's2

Parkinson's2

Sub Nigra
6.3
5.3
BA4
54.0
3.2

Control2

Parkinson's

Sub Nigra
58.6
10.2
BA4
6.2
0.0

Control

Alzheimer's2

BA17
39.2
9.3
BA4 Control2
0.0
4.2

Depression2

BA17 Depression
43.5
50.7
BA4 Control
35.6
4.7

[0805] CNS_neurodegeneration_v1.0 Summary: Ag2610/Ag2607/Ag2377 The CG50303-03 gene is expressed more highly in the temporal cortex of Alzheimer's diseased brain than in control brain without amyloid plaques, which are diagnostic and potentially causative of Alzheimer's disease. The CG50303-03 gene encodes a protein with homology to GPCRs.

[0806] GPCRs are readily targetable with drugs, and regulate many specific brain processes, including signaling processes, that are currently the target of FDA-approved pharmaceuticals that treat Alzheimer's disease, such as the cholinergic system. The major mechanisms proposed for AbetaP-induced cytotoxicity involve the loss of Ca2+ homeostasis and the generation of reactive oxygen species (ROS). The changes in Ca2+ homeostasis could be the result of changes in G-protein-driven releases of second messengers. Thus, targeting this class of molecule can have therapeutic potential in Alzheimer's disease treatment. In particular, the increased CG50303-03 gene expression in brains affected by Alzheimer's indicates potential therapeutic value to drugs that target this GPCR (Perrine et al., Cognitive functioning after pallidotomy for refractory Parkinson's disease. J Neurol Neurosurg Psychiatry 65(2): 150-4, 1998; Kourie, Mechanisms of amyloid beta protein-induced modification in ion transport systems: implications for neurodegenerative diseases. Cell Mol Neurobiol 21(3): 173-213, 2001).

[0807] Panel 1.2 Summary: Ag1501 The CG50303-03 gene is expressed at moderate levels throughout many of the samples in this panel. Highest expression is detected in an ovarian cancer cell line (CT=30.7). In addition, this gene is overexpressed in all six ovarian cancer cell lines present in this panel when compared to expression in normal ovary. The CG50303-03 gene is also moderately expressed in cell lines derived from melanoma, breast cancer, and lung cancer. Thus, the expression of this gene could be used to distinguish these cell lines from other tissue samples. In addition, therapeutic modulation of the CG50303-03 gene or its protein product, through the use of small molecule drugs or antibodies, might be useful in the treatment of ovarian cancer, breast cancer, lung cancer or melanoma.

[0808] Among tissues involved in metabolic function, the CG50303-03 gene is moderately expressed in the adrenal gland, heart, skeletal muscle, and adult liver. Interestingly, CG50303-03 gene expression is much lower in fetal liver and heart tissues than in the corresponding adult tissues. Thus, expression of the CG50303-03 gene could be used to differentiate between adult and fetal tissues derived from the heart and liver. Furthermore, this gene or its protein product may be important in the pathogenesis and/or treatment of disease in any or all of the above-named tissues.

[0809] There is widespread moderate expression of the CG50303-03 gene across many of the samples derived from the CNS, including the amygdala, cerebellum, hippocampus, thalamus, cerebral cortex, and spinal cord. Please see CNS_neurodegeneration_panel_v1.0 summary for description of potential utility in the treatment of CNS disorders.

[0810] Panel 1.3D Summary: Ag2610/Ag2607/Ag1585/Ag2377 Expression of the CG50303-03 gene appears to be limited to tissues involved in central nervous system function on this panel. Specifically, low but significant expression is detected in the thalamus, substantia nigra, spinal cord and fetal brain. Ag2545 Expression of the CG50303-03 gene is low/undetectable (Ct values>35) in all samples on this panel (data not shown).

[0811] Panel 2.2 Summary: Ag2377/Ag2607 Two experiments with two different probe and primer sets both show highest expression of the CG50303-03 gene in a sample derived from a breast cancer sample (CTs=34-34.7). Thus, the expression of this gene could be used to distinguish breast cancer samples from other samples and as a diagnostic marker for the presence of breast cancer. Furthermore, therapeutic modulation of the CG50303-03 gene or the activity of its protein product, through the use of small molecule drugs or antibodies, might be effective in the treatment of breast cancer. Ag2610/Ag1585 Expression of the CG50303-03 gene is low/undetectable (Ct values>35) in all samples on this panel (data not shown).

[0812] Panel 4D Summary: Ag2607/Ag2377 Two experiments with two different probe and primer sets show the CG50303-03 gene is up regulated in LPS-stimulated monocytes (CTw=32-34). The putative GPCR encoded by this gene may therefore be involved in the activation of monocytes in their function as antigen-presenting cells. This suggests that antibodies or small molecule therapeutics that block the function of this membrane protein may be useful as anti-inflammatory therapeutics for the treatment of autoimmune and inflammatory diseases. Furthermore, antibodies or small molecule therapeutics that stimulate the function of this GPCR may be useful therapeutics for the treatment of immunosupressed individuals. Please note that data from one experiment with probe and primer set Ag2610 showed low/undetectable expression in all the samples on this panel (CTs>35). (Data not shown.) Data from one experiment with the probe and primer set Ag1585 is not included because the amp plot suggests that there were experimental difficulties with this run.

[0813] Panel CNS—1 Summary: Ag2377 Two experiments with the same probe and primer set produce results that are in very good agreement. Expression of the CG50303-03 gene is highest in the substantia nigra of a Huntington's disease patient, indicating that this gene may participate in the genetic dysregulation associated with the neurodegeneration that occurs in this brain region. The substantia nigra is also critical to the progression of Parkinson's disease neurodegeneration. Thus, pharmacological targeting of the GPCR encoded by the CG50303-03 gene may help counter this genetic dysregulation and contribute to the restoration of normal function in Huntington's disease as well as potentially Parkinson's disease patients. Pharmacological modulation of GPCR signaling systems is the mechanism by which powerful depression therapies, such as SSRIs, exert their effect. Please note that a third experiment with the probe and primer set Ag1585 showed low/undetectable expression in all the samples on this panel (CTs>35). (Data not shown.)

[0814] Panel CNS—1.1 Summary: Ag2377 In two experiments using the same probe and primer, highest expression of the CG50303-03 gene is seen in the cingulate gyrus of patients with parasupranuclear palsy PSP (CTs=32) and depression. This observation indicates that targeting this GPCR could have therapeutic value in the treatment of these diseases.

[0815] AP. CG153775-01/GMAP000723_A: Olfactory Receptor

[0816] Expression of gene CG153775-01 was assessed using the primer-probe sets Ag1516 and Ag2017, described in Tables APA and APB. Results of the RTQ-PCR runs are shown in Tables APC, APD, and APE.

190TABLE APAProbe Name Ag1516StartSEQ IDPrimersSequencesLengthPositionNO:Forward5′-atattattgccagcccttcct-3′21137376ProbeTET-5′-tgtatttcttccttgcctgcctgtca-3′-TAMRA26170377Reverse5′-ttgggagaaatggtagtggaat-3′22212378

[0817]

191

TABLE APB

Probe Name Ag2017

Start
SEQ ID

Primers
Sequences
Length
Position
NO:

Forward
5′-atattattgccagcccttcct-3′
21
137
379

Probe
TET-5′-tgtatttcttccttgcctgcctgtca-3′-TAMRA
26
170
380

Reverse
5′-atggtagtggaatatgcagcat-3′
22
203
381

[0818]

192

TABLE APC

Panel 1.2

Rel. Exp. (%) Ag1516, Run

Rel. Exp. (%) Ag1516, Run

Tissue Name
142098713
Tissue Name
142098713

Endothelial cells
0.0
Renal ca. 786-0
0.0

Heart (Fetal)
0.8
Renal ca. A498
0.0

Pancreas
0.0
Renal ca. RXF 393
0.0

Pancreatic ca. CAPAN 2
0.0
Renal ca. ACHN
0.0

Adrenal Gland
0.0
Renal ca. UO-31
0.9

Thyroid
0.0
Renal ca. TK-10
0.0

Salivary gland
0.0
Liver
0.0

Pituitary gland
0.0
Liver (fetal)
0.0

Brain (fetal)
0.0
Liver ca. (hepatoblast)
0.0

HepG2

Brain (whole)
0.0
Lung
0.0

Brain (amygdala)
0.0
Lung (fetal)
0.0

Brain (cerebellum)
0.0
Lung ca. (small cell) LX-1
0.0

Brain (hippocampus)
0.0
Lung ca. (small cell)
49.7

NCI-H69

Brain (thalamus)
0.0
Lung ca. (s.cell var.)
0.0

SHP-77

Cerebral Cortex
0.0
Lung ca. (large cell)NCI-
1.8

H460

Spinal cord
0.0
Lung ca. (non-sm.cell)
0.6

A549

glio/astro U87-MG
0.0
Lung ca. (non-s.cell)
0.0

NCI-H23

glio/astro U-118-MG
0.0
Lung ca. (non-s.cell)
1.8

HOP-62

astrocytoma SW1783
0.0
Lung ca. (non-s.cl) NCI-
0.0

H522

neuro*; met SK-N-AS
0.0
Lung ca. (squam.) SW
0.0

900

astrocytoma SF-539
0.7
Lung ca. (squam.) NCI-
0.0

H596

astrocytoma SNB-75
1.7
Mammary gland
0.0

glioma SNB-19
0.8
Breast ca.* (pl.ef) MCF-7
0.0

glioma U251
0.6
Breast ca.* (pl.ef) MDA-
0.0

MB-231

glioma SF-295
0.0
Breast ca.* (pl.ef) T47D
18.0

Heart
0.0
Breast ca. BT-549
0.0

Skeletal Muscle
0.0
Breast ca. MDA-N
0.8

Bone marrow
0.0
Ovary
0.0

Thymus
0.0
Ovarian ca. OVCAR-3
0.0

Spleen
0.0
Ovarian ca. OVCAR-4
0.0

Lymph node
0.0
Ovarian ca. OVCAR-5
100.0

Colorectal Tissue
5.8
Ovarian ca. OVCAR-8
5.9

Stomach
0.0
Ovarian ca. IGROV-1
0.0

Small intestine
0.0
Ovarian ca. (ascites) SK-
0.0

OV-3

Colon ca. SW480
0.0
Uterus
0.0

Colon ca.* SW620
0.0
Placenta
0.0

(SW480 met)

Colon ca. HT29
1.5
Prostate
0.0

Colon ca. HCT-116
0.0
Prostate ca.* (bone met)
0.0

PC-3

Colon ca. CaCo-2
0.0
Testis
0.2

Colon ca. Tissue
21.0
Melanoma Hs688(A).T
0.0

(ODO3866)

Colon ca. HCC-2998
0.0
Melanoma* (met)
8.4

Hs688(B).T

Gastric ca.* (liver met)
0.0
Melanoma UACC-62
0.0

NCI-N87

Bladder
0.0
Melanoma M14
27.7

Trachea
0.0
Melanoma LOX IMVI
0.0

Kidney
0.0
Melanoma* (met) SK-
0.0

MEL-5

Kidney (fetal)
0.0

[0819]

193

TABLE APD

Panel 1.3D

Rel. Exp. (%)
Rel. Exp. (%)

Rel. Exp. (%)
Rel. Exp. (%)

Ag2017, Run
Ag2017, Run

Ag2017, Run
Ag2017, Run

Tissue Name
148203726
165618877
Tissue Name
148203726
165618877

Liver
0.0
64.6
Kidney (fetal)
0.0
0.0

adenocarcinoma

Pancreas
0.0
0.0
Renal ca. 786-0
0.0
0.0

Pancreatic ca.
0.0
0.0
Renal ca. A498
0.0
0.0

CAPAN 2

Adrenal gland
0.0
0.0
Renal ca. RXF
0.0
100.0

393

Thyroid
0.0
0.0
Renal ca. ACHN
0.0
0.0

Salivary gland
0.0
0.0
Renal ca. UO-31
0.0
0.0

Pituitary gland
0.0
0.0
Renal ca. TK-10
5.5
0.0

Brain (fetal)
0.0
0.0
Liver
0.0
0.0

Brain (whole)
0.0
33.0
Liver (fetal)
0.0
0.0

Brain (amygdala)
0.0
0.0
Liver ca.
0.0
0.0

(hepatoblast)

HepG2

Brain (cerebellum)
0.0
0.0
Lung
2.0
0.0

Brain (hippocampus)
0.0
0.0
Lung (fetal)
0.0
0.0

Brain (substantia
0.0
0.0
Lung ca. (small
0.0
0.0

nigra)

cell) LX-1

Brain (thalamus)
0.0
47.3
Lung ca. (small
0.0
40.6

cell) NCI-H69

Cerebral Cortex
0.0
0.0
Lung ca. (s.cell
0.0
0.0

var.) SHP-77

Spinal cord
3.2
0.0
Lung ca. (large
0.0
37.6

cell)NCI-H460

glio/astro U87-MG
0.0
0.0
Lung ca. (non-
6.9
0.0

sm. cell) A549

glio/astro U-118-
6.3
0.0
Lung ca. (non-
0.0
0.0

MG

s. cell) NCI-H23

astrocytoma
0.0
0.0
Lung ca. (non-
8.8
0.0

SW1783

s.cell) HOP-62

neuro*; met SK-N-
0.0
0.0
Lung ca. (non-
0.0
0.0

AS

s.cl) NCI-H522

astrocytoma SF-539
0.0
80.1
Lung ca.
0.0
0.0

(squam.) SW 900

astrocytoma SNB-75
3.0
0.0
Lung ca.
0.0
0.0

(squam.) NCI-

H596

glioma SNB-19
3.9
0.0
Mammary gland
0.0
0.0

glioma U251
0.0
64.2
Breast ca.*
0.0
0.0

(pl.ef) MCF-7

glioma SF-295
0.0
0.0
Breast ca.*
0.0
0.0

(pl.ef) MDA-

MB-231

Heart (fetal)
0.0
0.0
Breast ca.*
0.0
52.1

(pl.ef) T47D

Heart
0.0
0.0
Breast ca. BT-
1.8
0.0

549

Skeletal muscle
0.9
0.0
Breast ca. MDA-N
0.7
0.0

(fetal)

Skeletal muscle
0.0
0.0
Ovary
0.0
0.0

Bone marrow
2.1
0.0
Ovarian ca.
0.0
52.1

OVCAR-3

Thymus
0.0
0.0
Ovarian ca.
0.0
0.0

OVCAR-4

Spleen
0.0
0.0
Ovarian ca.
0.0
0.0

OVCAR-5

Lymph node
0.0
0.0
Ovarian ca.
0.0
0.0

OVCAR-8

Colorectal
47.0
23.5
Ovarian ca.
0.0
0.0

IGROV-1

Stomach
0.0
0.0
Ovarian ca.*
0.0
0.0

(ascites) SK-OV-3

Small intestine
0.0
0.0
Uterus
0.0
0.0

Colon ca. SW480
0.0
0.0
Plancenta
0.0
0.0

Colon ca.*
0.0
0.0
Prostate
0.0
0.0

SW620(SW480 met)

Colon ca. HT29
0.7
0.0
Prostate ca.*
0.0
0.0

(bone met)PC-3

Colon ca. HCT-116
0.0
0.0
Testis
0.0
0.0

Colon ca. CaCo-2
100.0
88.3
Melanoma
0.0
0.0

Hs688(A).T

Colon ca.
0.0
0.0
Melanoma*
0.0
0.0

tissue(ODO3866)

(met)

Hs688(B).T

Colon ca. HCC-2998
0.0
0.0
Melanoma
0.0
0.0

UACC-62

Gastric ca.* (liver
0.0
0.0
Melanoma M14
0.0
0.0

met) NCI-N87

Bladder
0.0
0.0
Melanoma LOX
0.0
0.0

IMVI

Trachea
0.0
0.0
Melanoma*
0.0
0.0

(met) SK-MEL-5

Kidney
0.0
0.0
Adipose
0.0
0.0

[0820]

194

TABLE APE

Panel 2.2

Rel. Exp. (%) Ag2017,

Rel. Exp. (%) Ag2017,

Tissue Name
Run 174232810
Tissue Name
Run 174232810

Normal Colon
0.0
Kidney Margin (OD04348)
0.0

Colon cancer (OD06064)
0.0
Kidney malignant cancer
1.7

(OD06204B)

Colon Margin (OD06064)
0.0
Kidney normal adjacent
0.0

tissue (OD06204E)

Colon cancer (OD06159)
0.0
Kidney Cancer (OD04450-
0.0

01)

Colon Margin (OD06159)
0.0
Kidney Margin (OD04450-
0.0

03)

Colon cancer (OD06297-04)
0.0
Kidney Cancer 8120613
0.0

Colon Margin (OD06297-
0.0
Kidney Margin 8120614
0.0

015)

CC Gr.2 ascend colon
7.3
Kidney Cancer 9010320
0.0

(OD03921)

CC Margin (ODO3921)
0.0
Kidney Margin 9010321
0.0

Colon cancer metastasis
0.0
Kidney Cancer 8120607
0.0

(OD06104)

Lung Margin (OD06104)
100.0
Kidney Margin 8120608
0.0

Colon mets to lung
0.0
Normal Uterus
0.0

(OD04451-01)

Lung Margin (OD04451-02)
6.2
Uterine Cancer 064011
0.0

Normal Prostate
0.0
Normal Thyroid
0.0

Postate Cancer (OD04410)
0.0
Thyroid Cancer 064010
0.0

Prostate Margin (OD04410)
0.0
Thyroid Cancer A302152
0.0

Normal Ovary
0.0
Thyroid Margin A302153
0.0

Ovarian cancer (OD06283-
0.0
Normal Breast
0.0

03)

Ovarian Margin (OD06283-
0.0
Breast Cancer (OD04566)
5.8

07)

Ovarian Cancer 064008
10.8
Breast Cancer 1024
0.0

Ovarian cancer (OD06145)
0.0
Breast Cancer (OD04590-
8.8

01)

Ovarian Margin (OD06145)
0.0
Breast Cancer Mets
31.4

(OD04590-03)

Ovarian cancer (OD06455-
0.0
Breast Cancer Metastasis
0.0

03)

(OD04655-05)

Ovarian Margin (OD06455-
0.0
Breast Cancer 064006
0.0

07)

Normal Lung
14.3
Breast Cancer 9100266
4.8

Invasive poor diff. lung
0.0
Breast Margin 9100265
7.0

adeno (ODO4945-01

Lung Margin (ODO4945-03)
0.0
Breast Cancer A209073
0.0

Lung Malignant Cancer
0.0
Breast Margin A2090734
0.0

(OD03126)

Lung Margin (OD03126)
0.0
Breast cancer (OD06083)
1.3

Lung Cancer (OD05014A)
0.0
Breast cancer node
0.0

metastasis (OD06083)

Lung Margin (OD05014B)
0.0
Normal Liver
0.0

Lung cancer (OD06081)
0.0
Liver Cancer 1026
0.0

Lung Margin (OD06081)
0.0
Liver Cancer 1025
0.0

Lung Cancer (OD04237-01)
0.0
Liver Cancer 6004-T
0.0

Lung Margin (OD04237-02)
0.0
Liver Tissue 6004-N
0.0

Ocular Melanoma Metastasis
0.0
Liver Cancer 6005-T
0.0

Ocular Melanoma Margin
0.0
Liver Tissue 6005-N
0.0

(Liver)

Melanoma Metastasis
36.6
Liver Cancer 064003
0.0

Melanoma Margin (Lung)
0.0
Normal Bladder
0.0

Normal Kidney
0.0
Bladder Cancer 1023
0.0

Kidney Ca, Nuclear grade 2
0.0
Bladder Cancer A302173
0.0

(OD04338)

Kidney Margin (OD04338)
0.0
Normal Stomach
0.0

Kidney Ca Nuclear grade 1/2
0.0
Gastric Cancer 9060397
0.0

(OD04339)

Kidney Margin (OD04339)
0.0
Stomach Margin 9060396
0.0

Kidney Ca, Clear cell type
0.0
Gastric Cancer 9060395
0.0

(OD04340)

Kidney Margin (OD04340)
4.2
Stomach Margin 9060394
0.0

Kidney Ca, Nuclear grade 3
0.0
Gastric Cancer 064005
0.0

(OD04348)

[0821] Panel 1.2 Summary: Ag1516 Expression of the CG153775-01 gene appears to be expressed in some representative samples from cultured cell lines derived from melanoma, ovarian cancer, breast cancer and lung cancer. In addition, there is also expression detected in a sample derived from a primary colon cancer. Thus, the therapeutic modulation of the CG153775-01 gene, through the use of small molecule drugs or antibodies, might be of utility in the treatment of melanoma, ovarian cancer, breast cancer, lung cancer or colon cancer.

[0822] Panel 1.3D Summary: Ag2017 The expression profile of this gene was run in two independent experiments; there was some concordance between the runs. Of those concordant samples, the expression of this gene appears to be highest in a cell line derived from a colon cancer (CT=31). In addition, there is significant expression in a colorectal tissue sample (CT=32). These observations suggest that CG153775-01 gene expression may be specific for colonic tissue or cancers derived from such. Thus, the expression of this gene may be useful as a marker of colonic derived cells. In addition, therapeutic modulation of the CG153775-01 gene product, through down-regulation of activity by small molecule drugs or antibodies, may have therapeutic value in the treatment of colon cancer.

[0823] Panel 2.2 Summary: Ag2017 Expression of the CG153775-01 gene appears to be highest in a sample of normal lung tissue adjacent to a colon cancer metastasis (CT=33.6). Other samples that show expression are at levels too low to be reliable. Thus, the expression of this gene might be a marker of normal lung tissue response to metastatic cancer. In this context, the expression of the CG153775-01 gene might be facilitating the process of metastasis to the lung and hence inhibition of this gene product's activity, through the use of small molecule drugs or antibodies, might be of utility for the treatment of metastatic cancer to lung.

[0824] Panel 4D Summary: Ag2017 Expression of the CG153775-01 gene is low/undetectable (CT values>34.4) across the samples on this panel. (Data not shown.)

[0825] AQ. CG59725-02/GMAP001521_A and CG59725-03/GMAP002826-A: Olfactory Receptor

[0826] Expression of gene CG59725-02 and variant CG59725-03 was assessed using the primer-probe sets Ag1520, Ag1524 and Ag1627, described in Tables AQA, AQB and AQC. Results of the RTQ-PCR runs are shown in Tables AQD, AQE, and AQF.

195TABLE AQAProbe Name Ag1520StartSEQ IDPrimersSequencesLengthPositionNO:Forward5′-cagggctgtgtgatacaattct-3′22296382ProbeTET-5′-tgcaacatttgcaaccagtgactgtt-3′-TAMRA26325383Reverse5′-taaggatccactgccatcatag-3′22360384

[0827]

196

TABLE AQB

Probe Name Ag1524

Start
SEQ ID

Primers
Sequences
Length
Position
NO:

Forward
5′-cagggctgtgtgatacaattct-3′
22
296
385

Probe
TET-5′-tgcaacatttgcaaccagtgactgtt-3′-TAMRA
26
325
386

Reverse
5′-taaggatccactgccatcatag-3′
22
360
384

[0828]

197

TABLE AQC

Probe Name Ag1627

Start
SEQ ID

Primers
Sequences
Length
Position
NO:

Forward
5′-cagggctgtgtgatacaattct-3′
22
296
388

Probe
TET-5′-tgcaacatttgcaaccagtgactgtt-3′-TAMRA
26
325
389

Reverse
5′-taaggatccactgccatcatag-3′
22
360
390

[0829]

198

TABLE AQD

Panel 1.2

Rel. Exp. (%)
Rel. Exp. (%)

Rel. Exp. (%)
Rel. Exp. (%)

Ag1520, Run
Ag1524, Run

Ag1520, Run
Ag1524, Run

Tissue Name
141990587
142098792
Tissue Name
141990587
142098792

Endothelial cells
0.0
0.3
Renal ca. 786-0
0.0
0.0

Heart (Fetal)
3.2
1.6
Renal ca. A498
0.1
0.2

Pancreas
0.0
0.0
Renal ca. RXF
0.0
0.0

393

Pancreatic ca.
0.0
0.0
Renal ca. ACHN
0.0
0.1

CAPAN 2

Adrenal Gland
0.1
0.8
Renal ca. UO-31
1.4
0.7

Thyroid
0.0
0.0
Renal ca. TK-10
0.4
0.0

Salivary gland
0.3
0.4
Liver
18.3
17.6

Pituitary gland
0.0
0.0
Liver (fetal)
0.4
0.0

Brain (fetal)
0.0
0.0
Liver ca.
0.0
0.0

(hepatoblast)

HepG2

Brain (whole)
0.0
0.7
Lung
0.0
0.0

Brain (amygdala)
0.7
0.3
Lung (fetal)
0.8
0.5

Brain (cerebellum)
0.0
0.0
Lung ca. (small
0.0
0.0

cell) LX-1

Brain
0.6
1.3
Lung ca. (small
3.9
1.7

(hippocampus)

cell) NCI-H69

Brain (thalamus)
1.1
0.6
Lung ca. (s.cell
0.0
0.1

var.) SHP-77

Cerebral Cortex
0.1
1.1
Lung ca. (large
0.0
0.4

cell)NCI-H460

Spinal cord
1.0
1.1
Lung ca. (non-
1.1
0.9

sm. cell) A549

glio/astro U87-MG
0.0
0.1
Lung ca. (non-
0.0
0.2

s.cell) NCI-H23

glio/astro U-118-
0.2
0.1
Lung ca. (non-
1.5
0.9

MG

s.cell) HOP-62

astrocytoma
0.2
0.1
Lung ca. (non-
0.0
0.2

SW1783

s.cl) NCI-H522

neuro*; met SK-N-
0.9
0.3
Lung ca.
1.3
0.4

AS

(squam.) SW 900

astrocytoma SF-
0.6
0.1
Lung ca.
0.9
0.4

539

(squam.) NCI-

H596

astrocytoma SNB-
0.0
0.0
Mammary gland
0.4
0.0

75

glioma SNB-19
1.9
1.6
Breast ca.* (pl.ef)
0.0
0.0

MCF-7

glioma U251
0.1
0.1
Breast ca.* (pl.ef)
0.0
0.0

MDA-MB-231

glioma SF-295
0.0
0.0
Breast ca.* (pl.
3.4
1.6

ef) T47D

Heart
3.1
3.0
Breast ca. BT-
0.5
0.9

549

Skeletal Muscle
0.7
1.3
Breast ca. MDA-N
10.2
6.4

Bone marrow
0.0
0.0
Ovary
0.0
0.2

Thymus
0.0
0.0
Ovarian ca.
0.0
0.0

OVCAR-3

Spleen
100.0
100.0
Ovarian ca.
0.0
0.0

OVCAR-4

Lymph node
0.0
0.0
Ovarian ca.
12.3
6.6

OVCAR-5

Colorectal Tissue
0.6
0.9
Ovarian ca.
0.8
0.0

OVCAR-8

Stomach
0.0
0.1
Ovarian ca.
0.7
0.1

IGROV-1

Small intestine
0.7
0.8
Ovarian ca.
0.1
0.1

(ascites) SK-OV-3

Colon ca. SW480
0.0
0.0
Uterus
0.0
0.0

Colon ca.* SW620
0.0
0.0
Placenta
0.6
0.4

(SW480 met)

Colon ca. HT29
1.0
0.4
Prostate
0.0
0.1

Colon ca. HCT-
0.0
0.0
Prostate ca.*
0.1
0.7

116

(bone met) PC-3

Colon ca. CaCo-2
0.0
0.0
Testis
0.4
0.0

Colon ca. Tissue
1.6
1.5
Melanoma
0.0
0.3

(ODO3866)

Hs688(A).T

Colon ca. HCC-
0.0
0.0
Melanoma* (met)
0.6
0.4

2998

Hs688(B).T

Gastric ca.* (liver
0.3
0.0
Melanoma
1.7
1.0

met) NCI-N87

UACC-62

Bladder
6.2
4.1
Melanoma M14
3.7
2.9

Trachea
0.0
0.0
Melanoma LOX
0.0
0.1

IMVI

Kidney
2.9
1.3
Melanoma* (met)
0.7
0.4

SK-MEL-5

Kidney (fetal)
1.1
1.1

[0830]

199

TABLE AQE

Panel 1.3D

Rel. Exp. (%)
Rel. Exp. (%)

Rel. Exp. (%)
Rel. Exp. (%)

Ag1524, Run
Ag1627, Run

Ag1524, Run
Ag1627, Run

Tissue Name
147076910
165531487
Tissue Name
147076910
165531487

Liver
0.9
0.0
Kidney (fetal)
0.0
0.0

adenocarcinoma

Pancreas
0.0
0.0
Renal ca. 786-0
0.0
0.0

Pancreatic ca.
0.0
0.0
Renal ca. A498
1.4
0.0

CAPAN 2

Adrenal gland
0.0
0.0
Renal ca. RXF
0.0
0.0

393

Thyroid
0.0
0.0
Renal ca. ACHN
0.0
0.0

Salivary gland
0.0
0.0
Renal ca. UO-31
0.0
0.0

Pituitary gland
0.0
0.0
Renal ca. TK-10
0.0
0.0

Brain (fetal)
0.0
0.0
Liver
1.2
1.1

Brain (whole)
1.1
1.0
Liver (fetal)
0.0
0.0

Brain (amygdala)
0.0
2.0
Liver ca.
0.0
0.0

(hepatoblast)

HepG2

Brain (cerebellum)
0.0
4.9
Lung
0.6
1.1

Brain (hippocampus)
0.0
0.0
Lung (fetal)
1.7
0.0

Brain (substantia
0.0
0.0
Lung ca. (small
0.0
0.0

nigra)

cell) LX-1

Brain (thalamus)
0.0
2.1
Lung ca. (small
0.0
0.0

cell) NCI-H69

Cerebral Cortex
0.0
0.0
Lung ca. (s.cell
0.0
0.0

var.) SHP-77

Spinal cord
3.2
10.2
Lung ca. (large
0.0
0.0

cell)NCI-H460

glio/astro U87-MG
0.0
0.0
Lung ca. (non-
0.0
0.0

sm. cell) A549

glio/astro U-118-
0.0
0.0
Lung ca. (non-
0.0
0.0

MG

s.cell) NCI-H23

astrocytoma
0.8
0.0
Lung ca. (non-
0.0
0.0

SW1783

s.cell) HOP-62

neuro*; met SK-N-
0.0
0.0
Lung ca. (non-
0.0
0.0

AS

s.cl) NCI-H522

astrocytoma SF-539
0.0
0.0
Lung ca.
0.9
0.0

(squam.) SW 900

astrocytoma SNB-75
0.0
0.0
Lung ca.
0.0
0.0

(squam.) NCI-

H596

glioma SNB-19
0.0
0.0
Mammary gland
0.9
0.0

glioma U251
0.0
0.0
Breast ca.*
0.0
0.0

(pl.ef) MCF-7

glioma SF-295
0.0
0.0
Breast ca.*
0.0
0.0

(pl.ef) MDA-

MB-231

Heart (fetal)
0.0
0.0
Breast ca.*
0.0
0.0

(pl.ef) T47D

Heart
0.5
2.2
Breast ca. BT-
0.9
0.0

549

Skeletal muscle
7.0
2.5
Breast ca. MDA-N
2.8
0.0

(fetal)

Skeletal muscle
0.0
0.0
Ovary
0.0
0.0

Bone marrow
0.0
0.0
Ovarian ca.
0.0
0.0

OVCAR-3

Thymus
0.0
0.0
Ovarian ca.
0.0
0.0

OVCAR-4

Spleen
100.0
100.0
Ovarian ca.
1.8
0.0

OVCAR-5

Lymph node
0.0
1.4
Ovarian ca.
0.0
0.0

OVCAR-8

Colorectal
1.5
0.0
Ovarian ca.
0.0
0.0

IGROV-1

Stomach
0.0
0.0
Ovarian ca.*
0.0
0.0

(ascites) SK-OV-3

Small intestine
0.0
0.0
Uterus
0.0
0.0

Colon ca. SW480
0.0
0.0
Plancenta
3.2
0.0

Colon ca.*
0.0
0.0
Prostate
0.0
0.0

SW620(SW480 met)

Colon ca. HT29
0.0
0.0
Prostate ca.*
0.0
0.0

(bone met)PC-3

Colon ca. HCT-116
0.0
0.0
Testis
0.7
0.0

Colon ca. CaCo-2
0.0
0.0
Melanoma
0.0
0.0

Hs688(A).T

Colon ca.
0.0
0.0
Melanoma*
0.0
0.0

tissue(ODO3866)

(met)

Hs688(B).T

Colon ca. HCC-2998
0.0
0.0
Melanoma
0.0
0.0

UACC-62

Gastric ca.* (liver
0.0
0.0
Melanoma M14
0.0
0.0

met) NCI-N87

Bladder
1.6
0.0
Melanoma LOX
0.0
0.0

IMVI

Trachea
0.0
0.0
Melanoma*
0.0
0.0

(met) SK-MEL-5

Kidney
0.0
0.0
Adipose
0.0
0.0

[0831]

200

TABLE AQF

Panel 2D

Rel.
Rel.
Rel.

Rel.
Rel.
Rel.

Exp. (%)
Exp. (%)
Exp. (%)

Exp. (%)
Exp. (%)
Exp. (%)

Ag1520,
Ag1524,
Ag1524,

Ag1520,
Ag1524,
Ag1524,

Run
Run
Run
Tissue
Run
Run
Run

Tissue Name
145049825
145158073
147077017
Name
145049825
145158073
147077017

Normal Colon
0.0
27.7
12.3
Kidney
0.0
0.0
0.0

Margin

8120608

CC Well to
0.0
0.0
25.5
Kidney
0.0
0.0
0.0

Mod Diff

Cancer

(ODO3866)

8120613

CC Margin
22.7
19.2
0.0
Kidney
11.0
16.6
0.0

(ODO3866)

Margin

8120614

CC Gr.2
0.0
0.0
0.0
Kidney
11.6
12.6
0.0

rectosigmoid

Cancer

(ODO3868)

9010320

CC Margin
0.0
0.0
0.0
Kidney
0.0
0.0
0.0

(ODO3868)

Margin

9010321

CC Mod Diff
11.5
0.0
0.0
Normal
0.0
0.0
0.0

(ODO3920)

Uterus

CC Margin
0.0
0.0
0.0
Uterus
9.4
0.0
0.0

(ODO3920)

Cancer

064011

CC Gr.2
0.0
0.0
0.0
Normal
0.0
0.0
5.9

ascend colon

Thyroid

(ODO3921)

CC Margin
0.0
0.0
0.0
Thyroid
0.0
0.0
0.0

(ODO3921)

Cancer

064010

CC from
21.5
39.2
32.8
Thyroid
20.2
0.0
0.0

Partial

Cancer

Hepatectomy

A302152

(ODO4309)

Mets

Liver Margin
0.0
41.2
58.6
Thyroid
0.0
0.0
11.3

(ODO4309)

Margin

A302153

Colon mets to
0.0
0.0
0.0
Normal
12.5
0.0
5.7

lung

Breast

(OD04451-01)

Lung Margin
0.0
0.0
0.0
Breast
0.0
0.0
0.0

(OD04451-02)

Cancer

(OD04566)

Normal
0.0
0.0
0.0
Breast
6.0
0.0
21.6

Prostate 6546-1

Cancer

(OD04590-

01)

Prostate Cancer
0.0
0.0
0.0
Breast
0.0
0.0
0.0

(OD04410)

Cancer Mets

(OD04590-

03)

Prostate
0.0
0.0
0.0
Breast
16.0
0.0
6.3

Margin

Cancer

(OD04410)

Metastasis

(OD04655-

05)

Prostate Cancer
0.0
0.0
0.0
Breast
9.4
0.0
0.0

(OD04720-01)

Cancer

064006

Prostate
0.0
0.0
0.0
Breast
0.0
0.0
0.0

Margin

Cancer 1024

(OD04720-02)

Normal Lung
36.9
100.0
90.8
Breast
0.0
0.0
0.0

061010

Cancer

9100266

Lung Met to
0.0
0.0
12.0
Breast
0.0
0.0
0.0

Muscle

Margin

(ODO4286)

9100265

Muscle Margin
10.2
0.0
0.0
Breast
0.0
0.0
16.2

(ODO4286)

Cancer

A209073

Lung
0.0
0.0
0.0
Breast
0.0
0.0
11.8

Malignant

Margin

Cancer

A2090734

(OD03126)

Lung Margin
10.1
17.7
54.0
Normal
37.9
50.0
94.0

(OD03126)

Liver

Lung Cancer
0.0
0.0
0.0
Liver Cancer
0.0
29.3
12.8

(OD04404)

064003

Lung Margin
64.2
78.5
100.0
Liver Cancer
4.2
19.6
37.1

(OD04404)

1025

Lung Cancer
0.0
0.0
0.0
Liver Cancer
0.0
0.0
0.0

(OD04565)

1026

Lung Margin
9.9
26.6
37.4
Liver Cancer
9.6
35.4
11.8

(OD04565)

6004-T

Lung Cancer
0.0
0.0
0.0
Liver Tissue
0.0
0.0
0.0

(OD04237-01)

6004-N

Lung Margin
100.0
54.7
79.0
Liver Cancer
0.0
0.0
0.0

(OD04237-02)

6005-T

Ocular Mel
0.0
0.0
12.2
Liver Tissue
0.0
0.0
8.1

Met to Liver

6005-N

(ODO4310)

Liver Margin
22.8
56.6
36.6
Normal
8.4
0.0
53.6

(ODO4310)

Bladder

Melanoma
0.0
0.0
0.0
Bladder
23.2
0.0
12.9

Mets to Lung

Cancer 1023

(OD04321)

Lung Margin
0.0
58.2
17.8
Bladder
32.8
55.9
52.1

(OD04321)

Cancer

A302173

Normal Kidney
21.2
25.5
17.6
Bladder
0.0
0.0
0.0

Cancer

(OD04718-

01)

Kidney Ca,
0.0
0.0
13.6
Bladder
0.0
25.9
7.3

Nuclear grade

Normal

2 (OD04338)

Adjacent

(OD04718-

03)

Kidney Margin
0.0
19.9
0.0
Normal
0.0
0.0
0.0

(OD04338)

Ovary

Kidney Ca
0.0
0.0
0.0
Ovarian
0.0
0.0
0.0

Nuclear grade

Cancer

1/2 (OD04339)

064008

Kidney Margin
0.0
0.0
10.9
Ovarian
0.0
0.0
0.0

(OD04339)

Cancer

(OD04768-

07)

Kidney Ca,
9.2
0.0
24.8
Ovary
0.0
0.0
8.7

Clear cell type

Margin

(OD04340)

(OD04768-

08)

Kidney Margin
0.0
0.0
0.0
Normal
0.0
0.0
2.1

(OD04340)

Stomach

Kidney Ca,
0.0
0.0
0.0
Gastric
0.0
0.0
0.0

Nuclear grade

Cancer

3 (OD04348)

9060358

Kidney Margin
0.0
0.0
0.0
Stomach
0.0
0.0
0.0

(OD04348)

Margin

9060359

Kidney Cancer
0.0
0.0
0.0
Gastric
0.0
0.0
0.0

(OD04622-01)

Cancer

9060395

Kidney Margin
0.0
0.0
0.0
Stomach
0.0
0.0
12.5

(OD04622-03)

Margin

9060394

Kidney Cancer
0.0
0.0
0.0
Gastric
0.0
0.0
0.0

(OD04450-01)

Cancer

9060397

Kidney Margin
5.9
0.0
12.2
Stomach
0.0
0.0
0.0

(OD04450-03)

Margin

9060396

Kidney Cancer
0.0
0.0
0.0
Gastric
0.0
0.0
0.0

8120607

Cancer

064005

[0832] Panel 1.2 Summary: Ag1520/Ag1524 Two runs with the same probe and primer set show highest expression of the CG59725-02 gene in the spleen (CTs=26-28), an important site of secondary immune responses. Therefore, antibodies or small molecule therapeutics that block the function of this GPCR may be useful as anti-inflammatory therapeutics for the treatment of allergies, autoimmune diseases, and inflammatory diseases.

[0833] This gene also shows higher levels of expression in adult liver (CTs=28-20) than in fetal liver (CTs=35-40). Thus, expression of this gene could be used to differentiate between adult and fetal liver.

[0834] This gene also shows expression in clusters of cells from melanoma, breast and lung cancer cell lines. Thus, expression of this gene could be used to differentiate between these samples and other samples on this panel and as a marker to detect the presence of these cancers.

[0835] This gene represents a novel G-protein coupled receptor (GPCR) that also shows expression in the brain. The GPCR family of receptors contains a large number of neurotransmitter receptors, including the dopamine, serotonin, a and b-adrenergic, acetylcholine muscarinic, histamine, peptide, and metabotropic glutamate receptors. GPCRs are excellent drug targets in various neurologic and psychiatric diseases. All antipsychotics have been shown to act at the dopamine D2 receptor; similarly novel antipsychotics also act at the serotonergic receptor, and often the muscarinic and adrenergic receptors as well. While the majority of antidepressants can be classified as selective serotonin reuptake inhibitors, blockade of the 5-HT IA and a2 adrenergic receptors increases the effects of these drugs. The GPCRs are also of use as drug targets in the treatment of stroke. Blockade of the glutamate receptors may decrease the neuronal death resulting from excitotoxicity; further more the purinergic receptors have also been implicated as drug targets in the treatment of cerebral ischemia. The b-adrenergic receptors have been implicated in the treatment of ADHD with Ritalin, while the a-adrenergic receptors have been implicated in memory. Therefore this gene may be of use as a small molecule target for the treatment of any of the described diseases (El Yacoubi et al., Adenosine A2A receptor antagonists are potential antidepressants: evidence based on pharmacology and A2A receptor knockout mice. Br J Pharmacol 134(1):68-77, 2001; Blier, Pharmacology of rapid-onset antidepressant treatment strategies. Clin Psychiatry 62 Suppl 15:12-7, 2001; Tranquillini and Reggiani, Glycine-site antagonists and stroke. Expert Opin Investig Drugs 8(11):1837-1848, 1999; Monopoli et al., Blockade of adenosine A2A receptors by SCH 58261 results in neuroprotective effects in cerebral ischaemia in rats. Neuroreport 9(17):3955-9, 1998).

[0836] Panel 1.3D Summary: Ag1524/Ag1627 (identical sequence) Two experiments with same probe and primer show expression of the CG59725-02 gene limited to the spleen (CTs=30-31). This expression profile is in agreement with expression seen in Panel 1.2, The spleen is an important site of secondary immune responses. Therefore, antibodies or small molecule therapeutics that block the function of this GPCR may be useful as anti-inflammatory therapeutics for the treatment of allergies, autoimmune diseases, and inflammatory diseases.

[0837] Panel 2D Summary: Ag1520/Ag1524 Three experiments with the same probe and primer set show results that are in very good agreement, with highest expression of the CG59725-02 gene in normal lung tissue (CTs=32-34). Thus, expression of this gene could be used to differentiate normal lung tissue from other samples on this panel and in particular from malignant lung tissue. Furthermore, therapeutic modulation of the function or expression of the protein encoded by this gene may be effective in the treatment of lung cancer.

[0838] Panel 4D Summary: Ag1525/Ag1627 (identical sequence) Expression of the CG59725-02 gene is low/undet. in all samples on this panel (CT>35) (Data not shown.)

[0839] AR. CG53390-01/GM2557p19-A: Olfactory Receptor

[0840] Expression of gene CG53390-01 was assessed using the primer-probe sets Ag2015 and Ag1588, described in Tables ARA and ARB. Results of the RTQ-PCR runs are shown in Tables ARC, ARD, and ARE

201TABLE ARAProbe Name Ag2015StartSEQ IDPrimerSequencesLengthPositionNO:Forward5′-aagctctcctgtgcagatacct-3′22571391ProbeTET-5′-ctacgagatggcgctgtccacct-3′-TAMRA23597392Reverse5′-aaagagggagcattaggatcag-3′22628393

[0841]

202

TABLE ARB

Probe Name Ag1588

Start
SEQ ID

Primers
Sequences
Length
Position
NO:

Forward
5′-aagctctcctgtgcagatacct-3′
22
571
394

Probe
TET-5′-ctacgagatggcgctgtccacct-3′-TAMRA
23
597
395

Reverse
5′-aaagagggagcattaggatcag-3′
22
628
396

[0842]

203

TABLE ARC

Panel 1.3D

Rel.
Rel.
Rel.

Rel.
Rel.
Rel.

Exp. (%)
Exp. (%)
Exp. (%)

Exp. (%)
Exp. (%)
Exp. (%)

Ag1588,
Ag2015,
Ag2015,

Ag1588,
Ag2015,
Ag2015,

Run
Run
Run

Run
Run
Run

Tissue Name
165529897
147837991
152152893
Tissue Name
165529897
147837991
152152893

Liver
0.0
0.0
0.0
Kidney (fetal)
0.0
0.0
0.0

adenocarcinoma

Pancreas
0.0
0.0
0.0
Renal ca. 786-0
0.0
0.0
0.0

Pancreatic ca.
0.0
0.0
0.0
Renal ca.
0.0
0.0
0.0

CAPAN 2

A498

Adrenal gland
0.0
0.0
0.6
Renal ca. RXF
0.0
0.8
0.0

393

Thyroid
0.0
0.0
0.0
Renal ca.
0.0
0.0
1.2

ACHN

Salivary gland
0.0
0.0
0.0
Renal ca. UO-
0.0
0.0
0.0

31

Pituitary gland
0.0
0.0
0.0
Renal ca. TK-
0.0
0.0
1.2

10

Brain (fetal)
0.0
0.0
0.0
Liver
0.0
0.0
0.0

Brain (whole)
0.0
0.0
0.0
Liver (fetal)
0.0
0.0
0.0

Brain
0.0
0.0
0.0
Liver ca.
0.0
0.0
0.0

(amygdala)

(hepatoblast)

HepG2

Brain
0.0
0.0
0.0
Lung
0.0
0.3
0.0

(cerebellum)

Brain
0.0
0.0
0.0
Lung (fetal)
0.0
0.0
0.0

(hippocampus)

Brain
0.0
0.0
0.0
Lung ca.
0.0
0.0
0.0

(substantia

(small cell)

nigra)

LX-1

Brain
0.0
0.0
0.0
Lung ca.
0.0
0.0
0.0

(thalamus)

(small cell)

NCI-H69

Cerebral Cortex
0.0
0.0
2.6
Lung ca.
100.0
100.0
100.0

(s.cell var.)

SHP-77

Spinal cord
0.0
0.0
0.0
Lung ca.
0.0
0.0
0.0

(large

cell)NCI-

H460

glio/astro U87-
0.0
0.0
0.0
Lung ca. (non-
0.0
0.0
0.0

MG

sm. cell) A549

glio/astro U-
0.0
0.0
1.1
Lung ca. (non-
1.7
1.6
2.1

118-MG

s.cell) NCI-

H23

astrocytoma
1.4
0.3
0.0
Lung ca. (non-
0.0
0.0
0.0

SW1783

s.cell) HOP-62

neuro*; met
0.0
0.0
0.0
Lung ca. (non-
0.0
0.0
0.0

SK-N-AS

s.cl) NCI-

H522

astrocytoma SF-
0.0
0.0
0.0
Lung ca.
0.0
0.0
0.0

539

(squam.) SW

900

astrocytoma
0.0
0.0
1.1
Lung ca.
0.0
0.0
0.0

SNB-75

(squam.) NCI-

H596

glioma SNB-19
0.0
0.3
2.0
Mammary
0.0
0.0
1.3

gland

glioma U251
0.7
0.0
0.0
Breast ca.*
0.0
0.5
0.0

(pl.ef) MCF-7

glioma SF-295
0.0
0.0
0.0
Breast ca.*
0.0
0.0
0.0

(pl.ef) MDA-

MB-231

Heart (fetal)
0.0
0.0
0.0
Breast ca.*
0.0
0.4
0.0

(pl.ef) T47D

Heart
0.0
0.0
0.7
Breast ca. BT-
0.0
0.0
0.6

549

Skeletal muscle
0.0
0.0
0.6
Breast ca.
0.0
0.9
0.0

(fetal)

MDA-N

Skeletal muscle
0.0
0.0
0.0
Ovary
0.0
0.0
0.0

Bone marrow
0.0
0.0
0.0
Ovarian ca.
0.0
0.8
0.0

OVCAR-3

Thymus
0.0
0.3
0.0
Ovarian ca.
0.0
0.0
0.0

OVCAR-4

Spleen
0.8
0.0
0.0
Ovarian ca.
0.0
0.4
0.0

OVCAR-5

Lymph node
0.0
0.0
0.0
Ovarian ca.
0.7
0.0
0.0

OVCAR-8

Colorectal
0.8
0.7
2.1
Ovarian ca.
0.0
0.0
0.0

IGROV-1

Stomach
0.0
0.0
0.0
Ovarian ca.*
0.0
0.0
0.0

(ascites) SK-

OV-3

Small intestine
0.0
0.0
0.6
Uterus
0.0
0.4
0.0

Colon ca.
0.0
0.0
0.0
Plancenta
0.0
0.8
2.4

SW480

Colon ca.*
0.0
0.0
1.9
Prostate
0.0
0.0
0.0

SW620(SW480

met)

Colon ca. HT29
0.0
0.0
0.0
Prostate ca.*
0.0
0.0
0.0

(bone met)PC-3

Colon ca. HCT-
0.0
0.7
0.0
Testis
0.0
1.6
0.5

116

Colon ca.
0.0
0.0
38.7
Melanoma
0.0
0.0
0.0

CaCo-2

Hs688(A).T

Colon ca.
0.0
0.0
1.6
Melanoma*
0.0
0.0
0.0

tissue(ODO3866)

(met)

Hs688(B).T

Colon ca. HCC-
0.0
0.0
0.6
Melanoma
0.0
0.0
0.0

2998

UACC-62

Gastric ca.*
0.0
0.0
0.7
Melanoma
0.0
0.0
0.0

(liver met) NCI-

M14

N87

Bladder
0.0
0.0
0.0
Melanoma
0.0
0.0
1.1

LOX IMVI

Trachea
0.0
0.0
0.0
Melanoma*
0.0
0.0
1.4

(met) SK-

MEL-5

Kidney
0.0
0.0
0.0
Adipose
0.0
0.0
0.0

[0843]

204

TABLE ARD

Panel 2D

Rel. Exp. (%) Ag2015,

Rel. Exp. (%) Ag2015,

Tissue Name
Run 152152937
Tissue Name
Run 152152937

Normal Colon
10.6
Kidney Margin 8120608
0.0

CC Well to Mod Diff
32.5
Kidney Cancer 8120613
0.0

(ODO3866)

CC Margin (ODO3866)
1.1
Kidney Margin 8120614
25.9

CC Gr.2 rectosigmoid
20.6
Kidney Cancer 9010320
0.0

(ODO3868)

CC Margin (ODO3868)
0.0
Kidney Margin 9010321
0.0

CC Mod Diff (ODO3920)
0.0
Normal Uterus
0.0

CC Margin (ODO3920)
0.0
Uterus Cancer 064011
0.0

CC Gr.2 ascend colon
0.0
Normal Thyroid
0.0

(ODO3921)

CC Margin (ODO3921)
48.6
Thyroid Cancer 064010
0.0

CC from Partial Hepatectomy
0.0
Thyroid Cancer A302152
0.0

(ODO4309) Mets

Liver Margin (ODO4309)
0.0
Thyroid Margin A302153
0.0

Colon mets to lung (OD04451-
0.0
Normal Breast
0.0

01)

Lung Margin (OD04451-02)
0.0
Breast Cancer (OD04566)
0.0

Normal Prostate 6546-1
0.0
Breast Cancer (OD04590-
0.0

01)

Prostate Cancer (OD04410)
0.0
Breast Cancer Mets
0.0

(OD04590-03)

Prostate Margin (OD04410)
0.0
Breast Cancer Metastasis
0.0

(OD04655-05)

Prostate Cancer (OD04720-01)
0.0
Breast Cancer 064006
0.0

Prostate Margin (OD04720-02)
0.0
Breast Cancer 1024
0.0

Normal Lung 061010
25.0
Breast Cancer 9100266
0.0

Lung Met to Muscle
0.0
Breast Margin 9100265
0.0

(ODO4286)

Muscle Margin (ODO4286)
0.0
Breast Cancer A209073
21.3

Lung Malignant Cancer
0.0
Breast Margin A2090734
9.7

(OD03126)

Lung Margin (OD03126)
0.0
Normal Liver
0.0

Lung Cancer (OD04404)
0.0
Liver Cancer 064003
0.8

Lung Margin (OD04404)
0.0
Liver Cancer 1025
0.0

Lung Cancer (OD04565)
0.0
Liver Cancer 1026
0.0

Lung Margin (OD04565)
0.0
Liver Cancer 6004-T
11.9

Lung Cancer (OD04237-01)
0.0
Liver Tissue 6004-N
100.0

Lung Margin (OD04237-02)
0.0
Liver Cancer 6005-T
0.0

Ocular Mel Met to Liver
0.0
Liver Tissue 6005-N
0.0

(ODO4310)

Liver Margin (ODO4310)
0.0
Normal Bladder
23.7

Melanoma Mets to Lung
0.0
Bladder Cancer 1023
19.9

(OD04321)

Lung Margin (OD04321)
0.0
Bladder Cancer A302173
60.7

Normal Kidney
0.0
Bladder Cancer
0.0

(OD04718-01)

Kidney Ca, Nuclear grade 2
0.0
Bladder Normal Adjacent
0.0

(OD04338)

(OD04718-03)

Kidney Margin (OD04338)
0.0
Normal Ovary
0.0

Kidney Ca Nuclear grade 1/2
0.0
Ovarian Cancer 064008
0.0

(OD04339)

Kidney Margin (OD04339)
0.0
Ovarian Cancer
26.1

(OD04768-07)

Kidney Ca, Clear cell type
0.0
Ovary Margin (OD04768-
0.0

(OD04340)

08)

Kidney Margin (OD04340)
0.0
Normal Stomach
0.0

Kidney Ca, Nuclear grade 3
0.0
Gastric Cancer 9060358
21.0

(OD04348)

Kidney Margin (OD04348)
0.0
Stomach Margin 9060359
0.0

Kidney Cancer (OD04622-01)
0.0
Gastric Cancer 9060395
0.0

Kidney Margin (OD04622-03)
0.0
Stomach Margin 9060394
0.0

Kidney Cancer (OD04450-01)
0.0
Gastric Cancer 9060397
0.0

Kidney Margin (OD04450-03)
24.7
Stomach Margin 9060396
0.0

Kidney Cancer 8120607
0.0
Gastric Cancer 064005
19.8

[0844]

205

TABLE ARE

Panel 4D

Rel.
Rel.
Rel.

Rel.
Rel.
Rel.

Exp. (%)
Exp. (%)
Exp. (%)

Exp. (%)
Exp. (%)
Exp. (%)

Ag1588,
Ag2015,
Ag2015,

Ag1588,
Ag2015,
Ag2015,

Run
Run
Run

Run
Run
Run

Tissue Name
165373604
152153145
152685551
Tissue Name
165373604
152153145
152685551

Secondary Th1 act
0.0
0.0
40.1
HUVEC IL-
0.0
0.0
0.0

1beta

Secondary Th2 act
26.4
25.3
26.1
HUVEC IFN
0.0
0.0
0.0

gamma

Secondary Tr1 act
12.3
58.6
0.0
HUVEC TNF
0.0
0.0
0.0

alpha + IFN

gamma

Secondary Th1
0.0
11.5
0.0
HUVEC TNF
0.0
0.0
0.0

rest

alpha + IL4

Secondary Th2
16.6
43.8
31.0
HUVEC IL-11
0.0
0.0
0.0

rest

Secondary Tr1 rest
42.6
24.5
14.2
Lung
0.0
0.0
0.0

Microvascular

EC none

Primary Th1 act
0.0
17.3
25.0
Lung
0.0
0.0
0.0

Microvascular

EC TNF alpha +

IL-1beta

Primary Th2 act
0.0
0.0
0.0
Microvascular
0.0
0.0
0.0

Dermal EC none

Primary Tr1 act
0.0
27.5
0.0
Microsvasular
0.0
0.0
24.0

Dermal EC

TNF alpha + IL-

1beta

Primary Th1 rest
18.6
69.7
12.3
Bronchial
0.0
0.0
0.0

epithelium

TNF alpha +

IL1beta

Primary Th2 rest
13.9
31.6
0.0
Small airway
0.0
0.0
0.0

epithelium none

Primary Tr1 rest
0.0
0.0
0.0
Small airway
0.0
0.0
0.0

epithelium

TNF alpha + IL-

1beta

CD45RA CD4
0.0
0.0
12.6
Coronery artery
0.0
44.4
0.0

lymphocyte act

SMC rest

CD45RO CD4
23.0
0.0
0.0
Coronery artery
0.0
0.0
0.0

lymphocyte act

SMC TNF alpha

+ IL-1beta

CD8 lymphocyte
0.0
0.0
0.0
Astrocytes rest
0.0
0.0
27.5

act

Secondary CD8
0.0
0.0
0.0
Astrocytes
0.0
0.0
0.0

lymphocyte rest

TNF alpha + IL-

1beta

Secondary CD8
27.5
0.0
0.0
KU-812
0.0
0.0
0.0

lymphocyte act

(Basophil) rest

CD4 lymphocyte
13.1
0.0
13.2
KU-812
0.0
0.0
41.8

none

(Basophil)

PMA/ionomycin

2ry
0.0
23.2
29.5
CCD1106
0.0
0.0
0.0

Th1/Th2/Tr1_anti-

(Keratinocytes)

CD95 CH11

none

LAK cells rest
15.2
12.0
0.0
CCD1106
0.0
0.0
24.7

(Keratinocytes)

TNF alpha + IL-

1beta

LAK cells IL-2
0.0
31.6
0.0
Liver cirrhosis
100.0
100.0
100.0

LAK cells IL-
16.6
33.2
0.0
Lupus kidney
0.0
0.0
0.0

2 + IL-12

LAK cells IL-
0.0
0.0
15.6
NCI-H292 none
0.0
0.0
0.0

2 + IFN gamma

LAK cells IL-2 +
0.0
0.0
25.3
NCI-H292 IL-4
0.0
26.6
0.0

IL-18

LAK cells
0.0
0.0
29.7
NCI-H292 IL-9
0.0
0.0
0.0

PMA/ionomycin

NK Cells IL-2 rest
35.6
25.5
20.3
NCI-H292 IL-13
0.0
0.0
0.0

Two Way MLR 3
0.0
0.0
0.0
NCI-H292 IFN
0.0
23.3
0.0

day

gamma

Two Way MLR 5
0.0
0.0
24.1
HPAEC none
0.0
0.0
0.0

day

Two Way MLR 7
0.0
0.0
0.0
HPAEC TNF
0.0
0.0
0.0

day

alpha + IL-1beta

PBMC rest
0.0
12.3
0.0
Lung fibroblast
0.0
14.2
0.0

none

PBMC PWM
10.1
0.0
0.0
Lung fibroblast
0.0
0.0
0.0

TNF alpha + IL-

1beta

PBMC PHA-L
0.0
0.0
24.3
Lung fibroblast
0.0
25.7
0.0

IL-4

Ramos (B cell)
0.0
0.0
0.0
Lung fibroblast
0.0
0.0
0.0

none

IL-9

Ramos (B cell)
0.0
16.3
0.0
Lung fibroblast
0.0
19.3
0.0

ionomycin

IL-13

B lymphocytes
0.0
13.7
43.2
Lung fibroblast
0.0
0.0
0.0

PWM

IFN gamma

B lymphocytes
0.0
20.2
0.0
Dermal
0.0
0.0
30.1

CD40L and IL-4

fibroblast

CCD1070 rest

EOL-1 dbcAMP
0.0
0.0
0.0
Dermal
57.4
40.9
25.7

fibroblast

CCD1070 TNF

alpha

EOL-1 dbcAMP
0.0
11.0
0.0
Dermal
0.0
0.0
0.0

PMA/ionomycin

fibroblast

CCD1070 IL-1

beta

Dendritic cells
0.0
0.0
0.0
Dermal
0.0
0.0
0.0

none

fibroblast IFN

gamma

Dendritic cells
0.0
0.0
0.0
Dermal
0.0
0.0
0.0

LPS

fibroblast IL-4

Dendritic cells
0.0
22.5
0.0
IBD Colitis 2
0.0
0.0
0.0

anti-CD40

Monocytes rest
0.0
0.0
23.2
IBD Crohn's
0.0
0.0
24.8

Monocytes LPS
0.0
0.0
22.7
Colon
0.0
0.0
29.9

Macrophages rest
0.0
0.0
24.5
Lung
0.0
22.5
56.3

Macrophages LPS
0.0
0.0
0.0
Thymus
14.7
0.0
0.0

HUVEC none
0.0
18.7
0.0
Kidney
0.0
0.0
0.0

HUVEC starved
0.0
0.0
0.0

[0845] CNS_neurodegeneration_v1.0 Summary: Ag2015 Expression of the CG53390-01 gene is low/undetectable.(CT>35) in all samples in this panel. (Data not shown)

[0846] Panel 1.3D Summary: Ag1588/Ag2015 (identical sequences) Three experiments with the same probe and primer set produce results that are in excellent agreement, with highest expression of the CG53390-01 gene in a lung cancer cell line (CTs=29). Expression of this gene is almost exclusive to this sample and thus, could potentially be used to differentiate between this sample and other samples on this panel and between normal and cancerous lung tissue. Furthermore, therapeutic modulation of the expression or function of this gene product may be effective in the treatment of lung cancer.

[0847] Panel 2D Summary: Ag2015 Highest expression of the CG53390-01 gene is seen in normal liver tissue (CT=33). Expression of this gene is almost exclusive to this sample and thus, could potentially be used to differentiate between this sample and other samples on this panel and between normal and cancerous liver tissue. Furthermore, therapeutic modulation of the expression or function of this gene product may be effective in the treatment of liver cancer.

[0848] Panel 4D Summary: Ag1588/Ag2015 Three experiments with the same probe and primer set show highest expression in liver cirrhosis (CTs=32-34). Furthermore, expression of this gene is not detected in normal liver in Panel 1.3D, suggesting that its expression is unique to liver cirrhosis. This gene encodes a putative GPCR; therefore, antibodies or small molecule therapeutics could reduce or inhibit fibrosis that occurs in liver cirrhosis. In addition, antibodies to this putative GPCR could also be used for the diagnosis of liver cirrhosis.

[0849] AS. CG55881-03/GM656o22_A: Olfactory Receptor

[0850] Expression of gene CG55881-03 was assessed using the primer-probe sets Ag1523 and Ag1898, described in Tables ASA and ASB. Results of the RTQ-PCR runs are shown in Tables ASC, ASD, and ASE

206TABLE ASAProbe Name Ag1523StartSEQ IDPrimersSequencesLengthPositionNO:Forward5′-agggcaagttcatagctctgtt-3′22809397ProbeTET-5′-ctacaccgtagtcactcctgcgctga-3′-TAMRA26831398Reverse5′-cgtgttcctcagggtgtaaata-3′22864399

[0851]

207

TABLE ASB

Probe Name Ag1898

Start
SEQ ID

Primers
Sequences
Length
Position
NO:

Forward
5′-ggctgtggtgtctctgttttac-3′
22
735
400

Probe
TET-5′-catcttcatgtatctccagccagcca-3′
26
765
401

Reverse
5′-ctatgaacttgccctgctcat-3′
21
803
402

[0852]

208

TABLE ASC

Panel 1.2

Rel. Exp. (%) Ag1523, Run

Rel. Exp. (%) Ag1523, Run

Tissue Name
142131146
Tissue Name
142131146

Endothelial cells
0.0
Renal ca. 786-0
0.0

Heart (Fetal)
0.0
Renal ca. A498
0.3

Pancreas
0.2
Renal ca. RXF 393
0.0

Pancreatic ca. CAPAN 2
0.0
Renal ca. ACHN
0.0

Adrenal Gland
0.2
Renal ca. UO-31
0.3

Thyroid
0.0
Renal ca. TK-10
0.2

Salivary gland
0.2
Liver
0.0

Pituitary gland
0.0
Liver (fetal)
0.1

Brain (fetal)
0.0
Liver ca. (hepatoblast)
0.0

HepG2

Brain (whole)
0.0
Lung
0.0

Brain (amygdala)
0.1
Lung (fetal)
0.0

Brain (cerebellum)
2.0
Lung ca. (small cell) LX-1
0.1

Brain (hippocampus)
0.1
Lung ca. (small cell)
2.4

NCI-H69

Brain (thalamus)
0.2
Lung ca. (s.cell var.)
0.1

SHP-77

Cerebral Cortex
0.0
Lung ca. (large cell)NCI-
0.6

H460

Spinal cord
0.0
Lung ca. (non-sm.cell)
1.0

A549

glio/astro U87-MG
0.0
Lung ca. (non-s.cell)
0.0

NCI-H23

glio/astro U-118-MG
0.0
Lung ca. (non-s.cell)
0.6

HOP-62

astrocytoma SW1783
0.1
Lung ca. (non-s.cl) NCI-
0.0

H522

neuro*; met SK-N-AS
0.0
Lung ca. (squam.) SW
0.2

900

astrocytoma SF-539
0.0
Lung ca. (squam.) NCI-
1.0

H596

astrocytoma SNB-75
0.2
Mammary gland
0.1

glioma SNB-19
0.4
Breast ca.* (pl.ef) MCF-7
0.0

glioma U251
0.0
Breast ca.* (pl.ef) MDA-
0.1

MB-231

glioma SF-295
0.0
Breast ca.* (pl.ef) T47D
0.6

Heart
0.3
Breast ca. BT-549
0.0

Skeletal Muscle
0.2
Breast ca. MDA-N
10.4

Bone marrow
0.2
Ovary
0.0

Thymus
0.0
Ovarian ca. OVCAR-3
0.0

Spleen
0.0
Ovarian ca. OVCAR-4
0.0

Lymph node
0.0
Ovarian ca. OVCAR-5
1.1

Colorectal Tissue
0.2
Ovarian ca. OVCAR-8
0.3

Stomach
0.0
Ovarian ca. IGROV-1
0.4

Small intestine
0.1
Ovarian ca. (ascites) SK-
0.4

OV-3

Colon ca. SW480
0.0
Uterus
0.0

Colon ca.* SW620
0.0
Placenta
0.1

(SW480 met)

Colon ca. HT29
0.2
Prostate
19.3

Colon ca. HCT-116
0.0
Prostate ca.* (bone met)
0.3

PC-3

Colon ca. CaCo-2
0.0
Testis
2.3

Colon ca. Tissue
0.8
Melanoma Hs688(A).T
0.0

(ODO3866)

Colon ca. HCC-2998
0.0
Melanoma* (met)
0.4

Hs688(B).T

Gastric ca.* (liver met)
0.1
Melanoma UACC-62
0.6

NCI-N87

Bladder
0.5
Melanoma M14
4.1

Trachea
0.0
Melanoma LOX IMVI
0.2

Kidney
0.1
Melanoma* (met) SK-
100.0

MEL-5

Kidney (fetal)
0.2

[0853]

209

TABLE ASD

Panel 1.3D

Rel. Exp. (%) Ag1898, Run

Rel. Exp. (%) Ag1898, Run

Tissue Name
165544705
Tissue Name
165544705

Liver adenocarcinoma
0.0
Kidney (fetal)
0.0

Pancreas
0.0
Renal ca. 786-0
0.0

Pancreatic ca. CAPAN 2
0.0
Renal ca. A498
0.0

Adrenal gland
0.0
Renal ca. RXF 393
0.0

Thyroid
0.0
Renal ca. ACHN
0.0

Salivary gland
0.0
Renal ca. UO-31
0.0

Pituitary gland
0.0
Renal ca. TK-10
0.0

Brain (fetal)
0.0
Liver
0.0

Brain (whole)
1.7
Liver (fetal)
0.0

Brain (amygdala)
0.0
Liver ca. (hepatoblast)
0.0

HepG2

Brain (cerebellum)
29.9
Lung
0.0

Brain (hippocampus)
0.0
Lung (fetal)
0.0

Brain (substantia nigra)
1.2
Lung ca. (small cell) LX-1
0.0

Brain (thalamus)
0.0
Lung ca. (small cell)
0.0

NCI-H69

Cerebral Cortex
0.0
Lung ca. (s.cell var.)
0.3

SHP-77

Spinal cord
0.0
Lung ca. (large cell)NCI-
0.0

H460

glio/astro U87-MG
0.0
Lung ca. (non-sm.cell)
0.0

A549

glio/astro U-118-MG
0.0
Lung ca. (non-s.cell)
0.0

NCI-H23

astrocytoma SW1783
0.0
Lung ca. (non-s.cell)
0.0

HOP-62

neuro*; met SK-N-AS
0.0
Lung ca. (non-s.cl) NCI-
0.0

H522

astrocytoma SF-539
2.6
Lung ca. (squam.) SW
0.0

900

astrocytoma SNB-75
0.0
Lung ca. (squam.) NCI-
0.0

H596

glioma SNB-19
0.0
Mammary gland
0.0

glioma U251
0.0
Breast ca.* (pl.ef) MCF-7
0.0

glioma SF-295
0.0
Breast ca.* (pl.ef) MDA-
0.0

MB-231

Heart (fetal)
0.0
Breast ca.* (pl.ef) T47D
0.0

Heart
0.0
Breast ca. BT-549
0.0

Skeletal muscle (fetal)
0.0
Breast ca. MDA-N
4.1

Skeletal muscle
0.0
Ovary
0.8

Bone marrow
0.0
Ovarian ca. OVCAR-3
0.0

Thymus
1.3
Ovarian ca. OVCAR-4
0.0

Spleen
0.0
Ovarian ca. OVCAR-5
0.0

Lymph node
2.2
Ovarian ca. OVCAR-8
0.0

Colorectal
0.0
Ovarian ca. IGROV-1
0.0

Stomach
0.0
Ovarian ca.* (ascites)
0.0

SK-OV-3

Small intestine
0.0
Uterus
0.0

Colon ca. SW480
0.0
Plancenta
0.0

Colon ca.* SW620(SW480
0.0
Prostate
4.2

met)

Colon ca. HT29
0.0
Prostate ca.* (bone
0.0

met)PC-3

Colon ca. HCT-116
0.0
Testis
31.9

Colon ca. CaCo-2
0.0
Melanoma Hs688(A).T
0.0

Colon ca.
0.0
Melanoma* (met)
0.0

tissue(ODO3866)

Hs688(B).T

Colon ca. HCC-2998
0.0
Melanoma UACC-62
2.3

Gastric ca.* (liver met)
0.0
Melanoma M14
6.0

NCI-N87

Bladder
0.0
Melanoma LOX IMVI
0.0

Trachea
0.0
Melanoma* (met) SK-
100.0

MEL-5

Kidney
0.0
Adipose
0.0

[0854]

210

TABLE ASE

Panel 2D

Rel. Exp. (%)
Rel. Exp. (%)

Rel. Exp. (%)
Rel. Exp. (%)

Ag1523, Run
Ag1523, Run

Ag1523, Run
Ag1523, Run

Tissue Name
145049949
145711220
Tissue Name
145049949
145711220

Normal Colon
5.2
0.0
Kidney Margin
0.0
0.0

8120608

CC Well to Mod
0.0
3.7
Kidney Cancer
0.0
0.0

Diff (ODO3866)

8120613

CC Margin
3.3
0.0
Kidney Margin
2.2
0.0

(ODO3866)

8120614

CC Gr.2
0.0
0.0
Kidney Cancer
0.0
0.0

rectosigmoid

9010320

(ODO3868)

CC Margin
0.0
4.2
Kidney Margin
0.0
0.0

(ODO3868)

9010321

CC Mod Diff
0.0
0.0
Normal Uterus
0.0
0.0

(ODO3920)

CC Margin
0.0
0.0
Uterus Cancer
0.0
0.0

(ODO3920)

064011

CC Gr.2 ascend
2.0
0.0
Normal Thyroid
0.0
0.0

colon (ODO3921)

CC Margin
0.0
0.0
Thyroid Cancer
0.0
0.0

(ODO3921)

064010

CC from Partial
0.0
0.0
Thyroid Cancer
0.0
0.0

Hepatectomy

A302152

(ODO4309) Mets

Liver Margin
0.0
2.0
Thyroid Margin
0.0
0.0

(ODO4309)

A302153

Colon mets to lung
0.0
0.0
Normal Breast
0.0
0.0

(OD04451-01)

Lung Margin
0.0
0.0
Breast Cancer
0.0
0.0

(OD04451-02)

(OD04566)

Normal Prostate
28.5
100.0
Breast Cancer
0.0
0.0

6546-1

(OD04590-01)

Prostate Cancer
100.0
56.3
Breast Cancer
0.0
0.0

(OD04410)

Mets (OD04590-

03)

Prostate Margin
23.5
27.2
Breast Cancer
0.0
0.0

(OD04410)

Metastasis

(OD04655-05)

Prostate Cancer
15.7
28.3
Breast Cancer
0.5
0.0

(OD04720-01)

064006

Prostate Margin
11.7
11.2
Breast Cancer
0.0
0.0

(OD04720-02)

1024

Normal Lung
0.0
4.8
Breast Cancer
5.0
0.0

061010

9100266

Lung Met to Muscle
0.0
0.0
Breast Margin
1.8
2.9

(ODO4286)

9100265

Muscle Margin
0.0
0.0
Breast Cancer
3.1
0.0

(ODO4286)

A209073

Lung Malignant
8.2
5.3
Breast Margin
0.0
0.0

Cancer (OD03126)

A2090734

Lung Margin
0.0
0.0
Normal Liver
0.0
0.0

(OD03126)

Lung Cancer
0.0
0.0
Liver Cancer
1.8
5.4

(OD04404)

064003

Lung Margin
1.6
0.0
Liver Cancer
0.0
0.0

(OD04404)

1025

Lung Cancer
0.0
0.0
Liver Cancer
0.0
0.0

(OD04565)

1026

Lung Margin
0.0
0.0
Liver Cancer
0.0
0.0

(OD04565)

6004-T

Lung Cancer
0.0
0.0
Liver Tissue
2.2
4.1

(OD04237-01)

6004-N

Lung Margin
0.0
0.0
Liver Cancer
4.0
0.0

(OD04237-02)

6005-T

Ocular Mel Met to
2.5
2.7
Liver Tissue
0.0
0.0

Liver (ODO4310)

6005-N

Liver Margin
0.0
0.0
Normal Bladder
0.0
0.0

(ODO4310)

Melanoma Mets to
3.6
0.0
Bladder Cancer
0.0
0.0

Lung (OD04321)

1023

Lung Margin
0.0
0.0
Bladder Cancer
0.8
5.8

(OD04321)

A302173

Normal Kidney
0.0
0.0
Bladder Cancer
0.0
0.0

(OD04718-01)

Kidney Ca, Nuclear
0.0
0.0
Bladder Normal
0.0
0.0

grade 2 (OD04338)

Adjacent

(OD04718-03)

Kidney Margin
0.0
0.0
Normal Ovary
0.0
0.0

(OD04338)

Kidney Ca Nuclear
0.0
0.0
Ovarian Cancer
0.0
0.0

grade 1/2

064008

(OD04339)

Kidney Margin
0.0
0.0
Ovarian Cancer
0.0
0.0

(OD04339)

(OD04768-07)

Kidney Ca, Clear
0.0
0.0
Ovary Margin
0.0
0.0

cell type (OD04340)

(OD04768-08)

Kidney Margin
0.0
0.0
Normal Stomach
0.0
0.0

(OD04340)

Kidney Ca, Nuclear
0.0
0.0
Gastric Cancer
0.0
0.0

grade 3 (OD04348)

9060358

Kidney Margin
0.0
4.6
Stomach Margin
0.0
0.0

(OD04348)

9060359

Kidney Cancer
0.0
0.0
Gastric Cancer
0.0
0.0

(OD04622-01)

9060395

Kidney Margin
0.0
0.0
Stomach Margin
3.7
0.0

(OD04622-03)

9060394

Kidney Cancer
0.0
0.0
Gastric Cancer
0.0
0.0

(OD04450-01)

9060397

Kidney Margin
0.0
0.0
Stomach Margin
0.0
0.0

(OD04450-03)

9060396

Kidney Cancer
0.0
0.0
Gastric Cancer
0.0
7.0

8120607

064005

[0855] Panel 1.2 Summary: Ag1523 Expression of the CG55881-03 gene is highest in a melanoma cancer cell line (CT=26.5), with expression detected in a cluster of melanoma cell lines. Thus, the expression of this gene could be used to distinguish samples derived from melanoma cell lines from other samples. In addition, therapeutic modulation of the expression or function of this gene, through the use of small molecule drugs or antibodies might be of use in the treatment of melanoma. In addition, the CG55881-03 gene is expressed in healthy prostate tissue but not in the prostate cancer cell line.

[0856] The CG55881-03 gene is also expressed differentially in the cerebellum. Cerebellar G protein function is known to be defective in Alzheimer's disease cerebella, suggesting this cerebellum-preferential GPCR may have utility as a drug target to counter the G-protein signaling deficit in Alzheimer's disease (Fowler et al., Receptor-effector coupling dysfunctions in Alzheimer's disease. Ann NY Acad. Sci. 786:294-304, 1996; Cowbum et al., Adenylyl cyclase activity in postmortem human brain: evidence of altered G protein mediation in Alzheimer's disease. J. Neurochem. 58:1409-19, 1992).

[0857] Panel 1.3D Summary: Ag1898 Highest expression of the CG55881-03 gene is detected in a melanoma cell line (CT=31) as is seen in Panel 1.2, with low but significant expression also seen in the cerebellum and testis. Thus, the expression of this gene could be used to distinguish samples derived from this melanoma cell line from other samples. In addition, therapeutic modulation of the expression or function of this gene, through the use of small molecule drugs or antibodies, might be of use in the treatment of melanoma. Please see Panel 1.2 summary for potential relevance of expression in cerebellum to the treatment of CNS disorders.

[0858] Panel 2D Summary: Ag1523 Two experiments with the same probe and primer set show expression of the CG55881-03 gene to be highest in a normal prostate in one run and a prostate cancer in the second run. In addition, the expression seen in both runs on panel 2D is specific to prostate derived samples. Thus, expression of the CG55881-03 gene could be used to distinguish samples derived from prostate tissue from other samples. Furthermore, since there is substantial over expression observed in a sample derived from prostate cancer when compared to a sample derived from its normal adjacent tissue, therapeutic modulation of the expression or function of the CG55881-03 gene product, through the use of small molecule drugs or antibodies, may be useful in the treatment of prostate cancer

[0859] Panel 4D Summary: Ag1898 Expression of the CG55881-03 gene is low/undetectable (Ct values>35) in all samples on this panel (data not shown).

[0860] AT. CG110025-02/GMbA144L1_A: Olfactory Receptor

[0861] Expression of gene CG110025-02 was assessed using the primer-probe sets Ag1370, Ag1621, Ag2437 and Ag2463, described in Tables ATA, ATB, ATC and ATD. Results of the RTQ-PCR runs are shown in Tables ATE, ATF, and ATG.

211TABLE ATAProbe Name Ag1370StartSEQ IDPrimersSequencesLengthPositionNO:Forward5′-atccttaccccattgttcaatc-3′22846403ProbeTET-5′-aaggaattcaaatcagccctacgaag-3′-TAMRA26891404Reverse5′-atagaaagtttggccgattgtt-3′22917405

[0862]

212

TABLE ATB

Probe Name Ag1621

Start
SEQ ID

Primers
Sequences
Length
Position
NO:

Forward
5′-atccttaccccattgttcaatc-3′
22
846
406

Probe
TET-5′-aaggaattcaaatcagccctacgaag-3′-TAMRA
26
891
407

Reverse
5′-atagaaagtttggccgattgtt-3′
22
917
408

[0863]

213

TABLE ATC

Probe Name Ag2437

Start
SEQ ID

Primers
Sequences
Length
Position
NO:

Forward
5′-accccattgttcaatccaat-3′
20
852
409

Probe
TET-5′-taaggaattcaaatcagccctacgaa-3′-TAMRA
26
890
410

Reverse
5′-gaaagtttggccgattgttc-3′
20
916
411

[0864]

214

TABLE ATD

Probe Name Ag2463

Start
SEQ ID

Primers
Senquences
Length
Position
NO:

Forward
5′-accccattgttcaatccaat-3′
20
852
412

Probe
TET-5′-taagaattcaaatcagccctacgaa-3′-TAMRA
26
890
413

Reverse
5′-gaaagtttggccgattgttc-3′
20
916
414

[0865]

215

TABLE ATE

Panel 1.2

Rel. Exp. (%) Ag1370, Run

Rel. Exp. (%) Ag1370, Run

Tissue Name
133467439
Tissue Name
133467439

Endothelial cells
0.0
Renal ca. 786-0
0.0

Heart (Fetal)
0.0
Renal ca. A498
0.0

Pancreas
0.0
Renal ca. RXF 393
0.0

Pancreatic ca. CAPAN 2
0.0
Renal ca. ACHN
0.0

Adrenal Gland
0.0
Renal ca. UO-31
0.0

Thyroid
0.0
Renal ca. TK-10
0.0

Salivary gland
0.0
Liver
0.0

Pituitary gland
0.0
Liver (fetal)
0.0

Brain (fetal)
0.0
Liver ca. (hepatoblast)
0.0

HepG2

Brain (whole)
0.0
Lung
0.0

Brain (amygdala)
0.0
Lung (fetal)
0.0

Brain (cerebellum)
0.0
Lung ca. (small cell) LX-1
0.0

Brain (hippocampus)
0.0
Lung ca. (small cell)
2.7

NCI-H69

Brain (thalamus)
0.0
Lung ca. (s.cell var.)
0.0

SHP-77

Cerebral Cortex
0.0
Lung ca. (large cell)NCI-
3.0

H460

Spinal cord
0.0
Lung ca. (non-sm.cell)
0.0

A549

glio/astro U87-MG
0.0
Lung ca. (non-s.cell)
0.0

NCI-H23

glio/astro U-118-MG
0.0
Lung ca. (non-s.cell)
0.0

HOP-62

astrocytoma SW1783
0.0
Lung ca. (non-s.cl) NCI-
0.0

H522

neuro*; met SK-N-AS
0.0
Lung ca. (squam.) SW
0.0

900

astrocytoma SF-539
0.0
Lung ca. (squam.) NCI-
0.0

H596

astrocytoma SNB-75
0.0
Mammary gland
0.0

glioma SNB-19
0.0
Breast ca.* (pl.ef) MCF-7
0.0

glioma U251
0.0
Breast ca.* (pl.ef) MDA-
0.0

MB-231

glioma SF-295
0.0
Breast ca.* (pl.ef) T47D
3.9

Heart
0.0
Breast ca. BT-549
1.7

Skeletal Muscle
0.0
Breast ca. MDA-N
0.0

Bone marrow
0.0
Ovary
0.0

Thymus
100.0
Ovarian ca. OVCAR-3
0.0

Spleen
0.0
Ovarian ca. OVCAR-4
0.0

Lymph node
0.0
Ovarian ca. OVCAR-5
1.2

Colorectal Tissue
0.0
Ovarian ca. OVCAR-8
0.0

Stomach
0.0
Ovarian ca. IGROV-1
0.0

Small intestine
0.0
Ovarian ca. (ascites) SK-
0.0

OV-3

Colon ca. SW480
0.0
Uterus
0.0

Colon ca.* SW620
0.0
Placenta
0.0

(SW480 met)

Colon ca. HT29
0.0
Prostate
0.0

Colon ca. HCT-116
0.0
Prostate ca.* (bone met)
0.0

PC-3

Colon ca. CaCo-2
0.0
Testis
0.0

Colon ca. Tissue
4.4
Melanoma Hs688(A).T
0.0

(ODO3866)

Colon ca. HCC-2998
0.0
Melanoma* (met)
0.0

Hs688(B).T

Gastric ca.* (liver met)
0.0
Melanoma UACC-62
0.0

NCI-N87

Bladder
1.2
Melanoma M14
3.3

Trachea
0.0
Melanoma LOX IMVI
0.0

Kidney
0.0
Melanoma* (met) SK-
0.0

MEL-5

Kidney (fetal)
0.0

[0866]

216

TABLE ATF

Panel 1.3D

Rel.
Rel.
Rel.

Rel.
Rel.
Rel.

Exp. (%)
Exp. (%)
Exp. (%)

Exp. (%)
Exp. (%)
Exp. (%)

Ag1621,
Ag2437,
Ag2463,

Ag1621,
Ag2437,
Ag2463,

Run
Run
Run

Run
Run
Run

Tissue Name
165531041
159555891
166108762
Tissue Name
165531041
159555891
166108762

Liver
0.0
0.0
0.0
Kidney
0.0
8.5
0.0

adenocarcinoma

(fetal)

Pancreas
0.0
0.0
0.0
Renal ca.
0.0
0.0
0.0

786-0

Pancreatic ca.
0.0
0.0
0.0
Renal ca.
0.0
0.0
0.0

CAPAN 2

A498

Adrenal gland
0.0
7.3
0.0
Renal ca.
0.0
0.0
0.0

RXF 393

Thyroid
0.0
0.0
0.0
Renal ca.
10.9
0.0
0.0

ACHN

Salivary gland
0.0
0.0
0.0
Renal ca.
0.0
0.0
0.0

UO-31

Pituitary gland
0.0
0.0
0.0
Renal ca.
0.0
0.0
5.4

TK-10

Brain (fetal)
0.0
8.4
0.0
Liver
0.0
0.0
0.0

Brain (whole)
0.0
0.0
0.0
Liver (fetal)
0.0
0.0
0.0

Brain (amygdala)
0.0
4.7
0.0
Liver ca.
0.0
0.0
8.7

(hepatoblast)

HepG2

Brain
0.0
0.0
0.0
Lung
0.0
0.0
0.0

(cerebellum)

Brain
0.0
3.9
0.0
Lung (fetal)
0.0
0.0
0.0

(hippocampus)

Brain (substantia
0.0
3.0
0.0
Lung ca.
0.0
0.0
0.0

nigra)

(small cell)

LX-1

Brain (thalamus)
0.0
0.0
0.0
Lung ca.
0.0
0.0
0.0

(small cell)

NCI-H69

Cerebral Cortex
0.0
0.0
0.0
Lung ca.
6.9
1.8
0.0

(s.cell var.)

SHP-77

Spinal cord
0.0
6.6
0.0
Lung ca.
0.0
2.7
0.0

(large

cell)NCI-

H460

glio/astro U87-
0.0
0.0
0.0
Lung ca.
0.0
0.0
0.0

MG

(non-sm. cell)

A549

glio/astro U-118-
0.0
0.0
0.0
Lung ca.
0.0
0.0
0.0

MG

(non-s.cell)

NCI-H23

astrocytoma
0.0
5.2
0.0
Lung ca.
0.0
1.5
0.0

SW1783

(non-s.cell)

HOP-62

neuro*; met SK-
0.0
0.0
0.0
Lung ca.
0.0
0.0
0.0

N-AS

(non-s.cl)

NCI-H522

astrocytoma SF-
0.0
0.0
0.0
Lung ca.
0.0
0.0
0.0

539

(squam.) SW

900

astrocytoma SNB-
0.0
4.7
0.0
Lung ca.
0.0
0.0
0.0

75

(squam.)

NCI-H596

glioma SNB-19
0.0
0.0
0.0
Mammary
0.0
6.0
0.0

gland

glioma U251
0.0
0.0
0.0
Breast ca.*
0.0
0.0
0.0

(pl.ef) MCF-7

glioma SF-295
0.0
0.0
0.0
Breast ca.*
0.0
0.0
0.0

(pl.ef) MDA-

MB-231

Heart (fetal)
0.0
0.0
0.0
Breast ca.*
0.0
0.0
0.0

(pl.ef) T47D

Heart
0.0
0.0
0.0
Breast ca.
0.0
0.0
14.3

BT-549

Skeletal muscle
0.0
0.0
0.0
Breast ca.
0.0
0.0
0.0

(fetal)

MDA-N

Skeletal muscle
10.2
0.0
0.0
Ovary
0.0
0.0
0.0

Bone marrow
0.0
0.0
0.0
Ovarian ca.
0.0
0.0
0.0

OVCAR-3

Thymus
100.0
100.0
100.0
Ovarian ca.
0.0
0.0
0.0

OVCAR-4

Spleen
0.0
0.0
0.0
Ovarian ca.
0.0
5.6
0.0

OVCAR-5

Lymph node
0.0
0.0
0.0
Ovarian ca.
0.0
0.0
0.0

OVCAR-8

Colorectal
0.0
1.5
0.0
Ovarian ca.
0.0
0.0
0.0

IGROV-1

Stomach
0.0
0.0
0.0
Ovarian ca.*
0.0
0.0
0.0

(ascites) SK-

OV-3

Small intestine
0.0
0.0
0.0
Uterus
3.4
7.2
0.0

Colon ca. SW480
0.0
0.0
0.0
Plancenta
0.0
0.0
0.0

Colon ca.*
0.0
0.0
0.0
Prostate
0.0
0.0
0.0

SW620(SW480

met)

Colon ca. HT29
0.0
0.0
0.0
Prostate ca.*
0.0
0.0
0.0

(bone

met)PC-3

Colon ca. HCT-
0.0
3.9
0.0
Testis
0.0
0.0
0.0

116

Colon ca. CaCo-2
0.0
10.5
0.0
Melanoma
0.0
0.0
0.0

Hs688(A).T

Colon ca.
0.0
0.0
0.0
Melanoma*
0.0
0.0
0.0

tissue(ODO3866)

(met)

Hs688(B).T

Colon ca. HCC-
0.0
0.0
0.0
Melanoma
0.0
0.0
0.0

2998

UACC-62

Gastric ca.* (liver
0.0
0.0
0.0
Melanoma
0.0
0.0
0.0

met) NCI-N87

M14

Bladder
0.0
0.0
0.0
Melanoma
12.1
0.0
0.0

LOX IMVI

Trachea
0.0
0.0
0.0
Melanoma*
0.0
0.0
0.0

(met) SK-

MEL-5

Kidney
0.0
0.0
0.0
Adipose
0.0
0.0
0.0

[0867]

217

TABLE ATG

Panel 4D

Rel.
Rel.
Rel.

Rel.
Rel.
Rel.

Exp. (%)
Exp. (%)
Exp. (%)

Exp. (%)
Exp. (%)
Exp. (%)

Ag1621,
Ag2437,
Ag2463,

Ag1621,
Ag2437,
Ag2463,

Run
Run
Run

Run
Run
Run

Tissue Name
164739992
159556063
164168557
Tissue Name
164739992
159556063
164168557

Secondary Th1 act
0.0
0.3
0.0
HUVEC IL-
0.0
0.0
0.0

1beta

Secondary Th2 act
0.0
0.0
0.0
HUVEC IFN
0.0
0.0
0.0

gamma

Secondary Tr1 act
0.0
0.0
0.0
HUVEC TNF
0.0
0.0
0.1

alpha + IFN

gamma

Secondary Th1
0.0
0.0
0.0
HUVEC TNF
0.0
0.7
0.0

rest

alpha + IL4

Secondary Th2
0.0
0.0
0.0
HUVEC IL-11
0.0
0.0
0.0

rest

Secondary Tr1 rest
0.0
0.0
0.0
Lung
0.0
0.0
0.0

Microvascular

EC none

Primary Th1 act
0.0
0.0
0.0
Lung
0.0
0.0
0.0

Microvascular

EC TNF alpha +

IL-1beta

Primary Th2 act
0.0
0.0
0.0
Microvascular
0.0
0.0
0.0

Dermal EC none

Primary Tr1 act
0.0
0.0
0.0
Microsvasular
0.0
0.0
0.0

Dermal EC

TNF alpha + IL-

1beta

Primary Th1 rest
0.0
0.0
0.0
Bronchial
0.0
0.0
0.0

epithelium

TNF alpha +

IL1beta

Primary Th2 rest
0.0
0.0
0.0
Small airway
0.0
0.6
0.0

epithelium none

Primary Tr1 rest
0.0
0.0
0.0
Small airway
0.0
0.0
0.0

epithelium

TNF alpha + IL-

1beta

CD45RA CD4
0.0
0.5
0.0
Coronery artery
0.0
0.0
0.0

lymphocyte act

SMC rest

CD45RO CD4
0.0
0.0
0.0
Coronery artery
0.0
0.6
0.0

lymphocyte act

SMC TNF

alpha + IL-1beta

CD8 lymphocyte
0.0
0.0
0.0
Astrocytes rest
0.0
0.0
0.0

act

Secondary CD8
0.0
0.0
0.0
Astrocytes
0.0
0.0
0.0

lymphocyte rest

TNF alpha + IL-

1beta

Secondary CD8
0.0
0.0
0.0
KU-812
0.0
0.6
0.0

lymphocyte act

(Basophil) rest

CD4 lymphocyte
0.0
0.0
0.0
KU-812
0.0
0.0
0.0

none

(Basophil)

PMA/ionomycin

2ry
0.0
1.2
0.0
CCD1106
0.0
0.0
0.0

Th1/Th2/Tr1_anti-

(Keratinocytes)

CD95 CH11

none

LAK cells rest
0.0
0.0
0.0
CCD1106
0.0
0.0
0.0

(Keratinocytes)

TNF alpha + IL-

1beta

LAK cells IL-2
0.0
0.0
0.0
Liver cirrhosis
2.2
2.1
0.6

LAK cells IL-
0.0
0.0
0.0
Lupus kidney
0.0
0.0
0.0

2 + IL-12

LAK cells IL-
0.0
0.0
0.0
NCI-H292 none
0.0
0.0
0.0

2 + IFN gamma

LAK cells IL-2 +
0.0
0.0
0.0
NCI-H292 IL-4
0.0
0.0
0.0

IL-18

LAK cells
0.0
0.0
0.0
NCI-H292 IL-9
0.0
0.0
0.0

PMA/ionomycin

NK Cells IL-2 rest
0.0
0.0
0.0
NCI-H292 IL-13
0.0
0.0
0.0

Two Way MLR 3
0.0
0.0
0.0
NCI-H292 IFN
0.0
0.0
0.0

day

gamma

Two Way MLR 5
0.0
0.0
0.0
HPAEC none
0.0
0.0
0.0

day

Two Way MLR 7
0.0
0.3
0.0
HPAEC TNF
0.0
0.0
0.0

day

alpha + IL-1beta

PBMC rest
0.0
0.0
0.0
Lung fibroblast
0.0
0.0
0.0

none

PBMC PWM
0.0
0.0
0.0
Lung fibroblast
0.0
0.7
0.0

TNF alpha + IL-

1beta

PBMC PHA-L
0.0
0.0
0.0
Lung fibroblast
0.0
0.0
0.0

IL-4

Ramos (B cell)
0.0
0.0
0.0
Lung fibroblast
0.0
0.0
0.0

none

IL-9

Ramos (B cell)
0.0
0.0
0.0
Lung fibroblast
0.0
0.0
0.0

ionomycin

IL-13

B lymphocytes
0.0
0.6
0.0
Lung fibroblast
0.0
0.6
0.0

PWM

IFN gamma

B lymphocytes
0.0
0.0
0.0
Dermal
0.0
0.0
0.0

CD40L and IL-4

fibroblast

CCD1070 rest

EOL-1 dbcAMP
0.0
0.0
0.0
Dermal
0.0
0.7
0.0

fibroblast

CCD1070 TNF

alpha

EOL-1 dbcAMP
0.0
0.4
0.0
Dermal
0.0
0.0
0.0

PMA/ionomycin

fibroblast

CCD1070 IL-1

beta

Dendritic cells
0.0
0.0
0.0
Dermal
0.0
0.0
0.0

none

fibroblast IFN

gamma

Dendritic cells
0.0
0.0
0.0
Dermal
0.8
0.5
0.2

LPS

fibroblast IL-4

Dendritic cells
0.0
0.0
0.0
IBD Colitis 2
0.0
0.7
0.3

anti-CD40

Monocytes rest
0.0
0.0
0.0
IBD Crohn's
2.1
0.2
0.1

Monocytes LPS
0.0
0.0
0.0
colon
0.0
0.3
0.0

Macrophages rest
0.0
0.0
0.0
Lung
0.5
0.0
0.0

Macrophages LPS
0.0
0.0
0.7
Thymus
0.0
0.0
0.3

HUVEC none
0.0
0.0
0.0
Kidney
100.0
100.0
100.0

HUVEC starved
0.0
0.0
0.0

[0868] Panel 1.2 Summary: Ag1370 Expression of the CG110025-02 gene in this panel is limited to the thymus (CT=33.2). CG110025-02 gene encodes a putative GPCR that may be expressed by cells normally present within these tissues. Alternatively, the transcript may be expressed by blood cells whose numbers may vary from tissue to tissue and may explain the discrepancy between panel 1.3 and 4D (see below). Modulation of the immune response by blocking the putative GPCR encoded for by the CG110025-02 gene could be important for organ transplant or controlling autoimmune diseases.

[0869] Panel 1.3D Summary: Ag1621/Ag2437Ag2463 Significant expression of the CG110025-02 gene is limited to thymus (CTs=33-34). This result is consistent with what is observed in Panel 1.2. The CG110025-02 gene encodes a putative GPCR that may be expressed by cells normally present within these tissues. Alternatively, the transcript may be expressed by blood cells whose numbers may vary from tissue to tissue and may explain the discrepancy between panel 1.3 and 4D (see below). Modulation of the immune response by blocking the putative GPCR encoded for by the CG110025-02 gene could be important for organ transplant or controlling autoimmune diseases.

[0870] Panel 2.2 Summary: Ag1621/Ag2463 Expression of the CG110025-02 gene is low/undetectable (CT values>35) across the samples on these panels. (Data not shown.)

[0871] Panel 2D Summary: Ag2437 Expression of the CG110025-02 gene is low/undetectable (CT values>35) across the samples on these panels. (Data not shown.)

[0872] Panel 4D Summary: Ag1621/Ag2437/Ag2463 The CG110025-02 transcript is only expressed at detectable levels in the kidney. The putative GPCR encoded for by the CG110025-02 gene could allow cells within the kidney to respond to specific microenvironmental signals (For example, ref. 1). Therefore, antibody or small molecule therapies designed with the protein encoded for by the CG110025-02 gene could modulate kidney function and be important in the treatment of inflammatory or autoimmune diseases that affect the kidney, including lupus and glomerulonephritis (Mark et al., G protein modulation of recombinant P/Q-type calcium channels by regulators of G protein signalling proteins. J. Physiol. 528 Pt 1: 65-77, 2000).

[0873] AU. CG155916-01/GMAP000435_A: Olfactory Receptor

[0874] Expression of gene CG155916-01 was assessed using the primer-probe set Ag2412, described in Table AUA. Results of the RTQ-PCR runs are shown in Table AUB.

218TABLE AUAProbe Name Ag2412StartSEQ IDPrimersSequencesLengthPositionNO:Forward5′-ttaatcctgctggactcatc-3′22162415ProbeTET-5′-tttctcagtaacctgtctcttgcagg-3′-TAMRA26204416Reverse5′-gacagctgaggagtaaccaatg-3′22230417

[0875]

219

TABLE AUB

Panel 4D

Rel. Exp. (%) Ag2412,

Rel. Exp. (%) Ag2412,

Tissue Name
Run 163869095
Tissue Name
Run 163869095

Secondary Th1 act
0.0
HUVEC IL-1 beta
0.0

Secondary Th2 act
0.0
HUVEC IFN gamma
0.0

Secondary Tr1 act
0.0
HUVEC TNF alpha + IFN
3.5

gamma

Secondary Th1 rest
0.0
HUVEC TNF alpha + IL4
3.9

Secondary Th2 rest
0.0
HUVEC IL-11
0.0

Secondary Tr1 rest
0.0
Lung Microvascular EC none
27.9

Primary Th1 act
0.0
Lung Microvascular EC
38.2

TNF alpha + IL-1 beta

Primary Th2 act
0.0
Microvascular Dermal EC none
100.0

Primary Tr1 act
0.0
Microsvasular Dermal EC
24.1

TNF alpha + IL-1 beta

Primary Th1 rest
0.0
Bronchial epithelium TNF alpha +
0.0

IL1 beta

Primary Th2 rest
0.0
Small airway epithelium none
0.0

Primary Tr1 rest
0.0
Small airway epithelium
0.0

TNF alpha + IL-1 beta

CD45RA CD4 lymphocyte
0.0
Coronery artery SMC rest
0.0

act

CD45RO CD4 lymphocyte
0.0
Coronery artery SMC TNF alpha +
10.9

act

IL-1 beta

CD8 lymphocyte act
0.0
Astrocytes rest
0.0

Secondary CD8
0.0
Astrocytes TNF alpha + IL-1 beta
4.7

lymphocyte rest

Secondary CD8
0.0
KU-812 (Basophil) rest
0.0

lymphocyte act

CD4 lymphocyte none
0.0
KU-812 (Basophil)
0.0

PMA/ionomycin

2ry Th1/Th2/Tr1_anti-
0.0
CCD1106 (Keratinocytes) none
0.0

CD95 CH11

LAK cells rest
0.0
CCD1106 (Keratinocytes)
0.0

TNF alpha + IL-1 beta

LAK cells IL-2
0.0
Liver cirrhosis
25.7

LAK cells IL-2 + IL-12
0.0
Lupus kidney
0.0

LAK cells IL-2 + IFN
0.0
NCI-H292 none
0.0

gamma

LAK cells IL-2 + IL-18
0.0
NCI-H292 IL-4
0.0

LAK cells
0.0
NCI-H292 IL-9
0.0

PMA/ionomycin

NK Cells IL-2 rest
0.0
NCI-H292 IL-13
0.0

Two Way MLR 3 day
0.0
NCI-H292 IFN gamma
0.0

Two Way MLR 5 day
0.0
HPAEC none
0.0

Two Way MLR 7 day
0.0
HPAEC TNF alpha + IL-1 beta
8.4

PBMC rest
0.0
Lung fibroblast none
0.0

PBMC PWM
0.0
Lung fibroblast TNF alpha + IL-
4.2

1 beta

PBMC PHA-L
1.7
Lung fibroblast IL-4
0.0

Ramos (B cell) none
0.0
Lung fibroblast IL-9
0.0

Ramos (B cell) ionomycin
0.0
Lung fibroblast IL-13
0.9

B lymphocytes PWM
0.0
Lung fibroblast IFN gamma
0.0

B lymphocytes CD40L
0.0
Dermal fibroblast CCD1070 rest
3.5

and IL-4

EOL-1 dbcAMP
0.0
Dermal fibroblast CCD1070
0.0

TNF alpha

EOL-1 dbcAMP
0.0
Dermal fibroblast CCD1070 IL-
0.0

PMA/ionomycin

1 beta

Dendritic cells none
0.0
Dermal fibroblast IFN gamma
0.0

Dendritic cells LPS
0.0
Dermal fibroblast IL-4
13.3

Dendritic cells anti-CD40
0.0
IBD Colitis 2
0.0

Monocytes rest
0.0
IBD Crohn's
0.0

Monocytes LPS
0.0
Colon
0.0

Macrophages rest
0.0
Lung
0.0

Macrophages LPS
0.0
Thymus
3.4

HUVEC none
3.2
Kidney
2.4

HUVEC starved
1.7

[0876] CNS_neurodegeneration_v1.0 Summary: Ag2412 Expression of the CG155916-01 gene is low/undetectable in all samples in this panel (CT>35). (Data not shown.)

[0877] Panel 1.3D Summary: Ag2412 Expression of the CG155916-01 gene is low/undetectable in all samples in this panel (CT>35). (Data not shown.)

[0878] Panel 2.2 Summary: Ag2412 Expression of the CG155916-01 gene is low/undetectable in all samples in this panel (CT>35). (Data not shown.)

[0879] Panel 4D Summary: Ag2412: The CG155916-01 transcript is most highly expressed in dermal microvasculature (CT=32.2), and to a lower extent in lung microvasculature. This expression profile suggests a role in angiogenesis and normal integrity of these tissues for the GPCR homolog encoded by this transcript. Therefore, agonistic protein therapeutics designed against this gene product may be beneficial for the improvement the effectiveness of skin transplantation. In addition, protein therapeutics designed with the putative GPCR encoded for by this protein could reduce or eliminate inflammation and tissue destruction due to inflammatory skin and lung diseases such as psoriasis, contact dermatitis, asthma, emphysema and chronic obstructive pulmonary diseases.

[0880] AV. CG155769-01/GMAC025942_B: Olfactory Receptor

[0881] Expression of gene CG155769-01 was assessed using the primer-probe set Ag2299, described in Table AVA. Results of the RTQ-PCR runs are shown in Tables AVB and AVC.

220TABLE AVAProbe Name Ag2299StartSEQ IDPrimersSequencesLengthPositionNO:Forward5′-aaagggatacgaaaagctgtct-3′22694418ProbeTET-5′-ccacctgtggggctcatctcttatct-3′-TAMRA26716419Reverse5′-cagagcccagatatttgaaggt-3′22766420

[0882]

221

TABLE AVB

Panel 1.3D

Rel. Exp. (%) Ag2299, Run

Rel. Exp. (%) Ag2299, Run

Tissue Name
151630259
Tissue Name
151630259

Liver adenocarcinoma
0.0
Kidney (fetal)
0.0

Pancreas
0.0
Renal ca. 786-0
0.0

Pancreatic ca. CAPAN 2
0.0
Renal ca. A498
0.0

Adrenal gland
0.0
Renal ca. RXF 393
0.0

Thyroid
0.0
Renal ca. ACHN
0.0

Salivary gland
0.0
Renal ca. UO-31
0.0

Pituitary gland
0.0
Renal ca. TK-10
0.0

Brain (fetal)
0.0
Liver
0.0

Brain (whole)
0.0
Liver (fetal)
0.0

Brain (amygdala)
0.0
Liver ca. (hepatoblast)
0.0

HepG2

Brain (cerebellum)
0.0
Lung
8.8

Brain (hippocampus)
0.0
Lung (fetal)
0.0

Brain (substantia nigra)
0.0
Lung ca. (small cell) LX-1
0.0

Brain (thalamus)
0.0
Lung ca. (small cell)
0.0

NCI-H69

Cerebral Cortex
0.0
Lung ca. (s.cell var.)
91.4

SHP-77

Spinal cord
0.0
Lung ca. (large cell)NCI-
0.0

H460

glio/astro U87-MG
0.0
Lung ca. (non-sm.cell)
0.0

A549

glio/astro U-118-MG
1.7
Lung ca. (non-s.cell)
0.0

NCI-H23

astrocytoma SW1783
0.0
Lung ca. (non-s.cell)
0.0

HOP-62

neuro*; met SK-N-AS
0.0
Lung ca. (non-s.cl) NCI-
0.0

H522

astrocytoma SF-539
0.0
Lung ca. (squam.) SW
0.0

900

astrocytoma SNB-75
0.0
Lung ca. (squam.) NCI-
0.0

H596

glioma SNB-19
0.0
Mammary gland
0.0

glioma U251
0.0
Breast ca.* (pl.ef) MCF-7
0.0

glioma SF-295
0.0
Breast ca.* (pl.ef) MDA-
0.0

MB-231

Heart (fetal)
0.0
Breast ca.* (pl.ef) T47D
0.0

Heart
0.0
Breast ca. BT-549
0.0

Skeletal muscle (fetal)
0.0
Breast ca. MDA-N
0.0

Skeletal muscle
0.0
Ovary
0.0

Bone marrow
0.0
Ovarian ca. OVCAR-3
0.0

Thymus
0.0
Ovarian ca. OVCAR-4
0.0

Spleen
0.0
Ovarian ca. OVCAR-5
0.0

Lymph node
0.0
Ovarian ca. OVCAR-8
0.0

Colorectal
100.0
Ovarian ca. IGROV-1
0.0

Stomach
0.0
Ovarian ca.* (ascites)
0.0

SK-OV-3

Small intestine
0.0
Uterus
0.0

Colon ca. SW480
0.0
Plancenta
0.0

Colon ca.* SW620(SW480
0.0
Prostate
0.0

met)

Colon ca. HT29
0.0
Prostate ca.* (bone
0.0

met)PC-3

Colon ca. HCT-116
0.0
Testis
62.4

Colon ca. CaCo-2
0.0
Melanoma Hs688(A).T
0.0

Colon ca.
0.0
Melanoma* (met)
0.0

tissue(ODO3866)
0.0
Hs688(B).T

Colon ca. HCC-2998
0.0
Melanoma UACC-62
0.0

Gastric ca.* (liver met)
0.0
Melanoma M14
0.0

NCI-N87

Bladder
0.0
Melanoma LOX IMVI
0.0

Trachea
0.0
Melanoma* (met) SK-
0.0

MEL-5

Kidney
0.0
Adipose
0.0

[0883]

222

TABLE AVC

Panel 4.1D

Rel. Exp. (%)
Rel. Exp. (%)

Rel. Exp. (%)
Rel. Exp. (%)

Ag2299, Run
Ag2299, Run

Ag2299, Run
Ag2299, Run

Tissue Name
169830289
229542251
Tissue Name
169830289
229542251

Secondary Th1 act
0.0
27.5
HUVEC IL-1beta
0.0
0.0

Secondary Th2 act
0.0
0.0
HUVEC IFN gamma
0.0
0.0

Secondary Tr1 act
0.0
0.0
HUVEC TNF alpha +
0.0
0.0

IFN gamma

Secondary Th1 rest
0.0
0.0
HUVEC TNF alpha +
0.0
0.0

IL4

Secondary Th2 rest
0.0
0.0
HUVEC IL-11
0.0
0.0

Secondary Tr1 rest
0.0
0.0
Lung Microvascular
0.0
0.0

EC none

Primary Th1 act
0.0
0.0
Lung Microvascular
0.0
0.0

EC TNF alpha + IL-

1beta

Primary Th2 act
0.0
0.0
Microvascular
0.0
0.0

Dermal EC none

Primary Tr1 act
0.0
0.0
Microvascular Dermal
0.0
0.0

EC TNF alpha + IL-

1beta

Primary Th1 rest
0.0
0.0
Bronchial epithelium
0.0
0.0

TNF alpha + IL1beta

Primary Th2 rest
0.0
0.0
Small airway
0.0
0.0

epithelium none

Primary Tr1 rest
0.0
0.0
Small airway
0.0
0.0

epithelium TNF

alpha + IL-1beta

CD45RA CD4
0.0
0.0
Coronery artery SMC
0.0
0.0

lymphocyte act

rest

CD45RO CD4
0.0
0.0
Coronery artery SMC
0.0
0.0

lymphocyte act

TNF alpha + IL-1beta

CD8 lymphocyte act
0.0
0.0
Astrocytes rest
0.0
0.0

Secondary CD8
0.0
0.0
Astrocytes TNF
0.0
0.0

lymphocyte rest

alpha + IL-1beta

Secondary CD8
0.0
0.0
KU-812 (Basophil)
0.0
0.0

lymphocyte act

rest

CD4 lymphocyte
0.0
0.0
KU-812 (Basophil)
0.0
0.0

none

PMA/ionomycin

2ry
0.0
0.0
CCD1106
0.0
0.0

Th1/Th2/Tr1_anti-

(Keratinocytes) none

CD95 CH11

LAK cells rest
0.0
0.0
CCD1106
0.0
0.0

(Keratinocytes)

TNF alpha + IL-1beta

LAK cells IL-2
0.0
0.0
Liver cirrhosis
0.0
0.0

LAK cells IL-2 + IL-
0.0
0.0
NCI-H292 none
0.0
0.0

12

LAK cells IL-2 + IFN
0.0
0.0
NCI-H292 IL-4
0.0
0.0

gamma

LAK cells IL-2 + IL-
0.0
43.2
NCI-H292 IL-9
0.0
0.0

18

LAK cells
0.0
0.0
NCI-H292 IL-13
0.0
0.0

PMA/ionomycin

NK Cells IL-2 rest
0.0
0.0
NCI-H292 IFN
0.0
0.0

gamma

Two Way MLR 3
0.0
36.1
HPAEC none
0.0
0.0

day

Two Way MLR 5
0.0
0.0
HPAEC TNF alpha +
0.0
0.0

day

IL-1beta

Two Way MLR 7
0.0
0.0
Lung fibroblast none
0.0
0.0

day

PBMC rest
0.0
0.0
Lung fibroblast TNF
0.0
0.0

alpha + IL-1beta

PBMC PWM
0.0
0.0
Lung fibroblast IL-4
0.0
0.0

PBMC PHA-L
0.0
0.0
Lung fibroblast IL-9
0.0
0.0

Ramos (B cell) none
0.0
0.0
Lung fibroblast IL-13
0.0
0.0

Ramos (B cell)
0.0
0.0
Lung fibroblast IFN
0.0
0.0

ionomycin

gamma

B lymphocytes PWM
0.0
0.0
Dermal fibroblast
0.0
0.0

CCD1070 rest

B lymphocytes
0.0
0.0
Dermal fibroblast
0.0
0.0

CD40L and IL-4

CCD1070 TNF alpha

EOL-1 dbcAMP
0.0
0.0
Dermal fibroblast
0.0
0.0

CCD1070 IL-1beta

EOL-1 dbcAMP
0.0
0.0
Dermal fibroblast IFN
0.0
0.0

PMA/ionomycin

gamma

Dendritic cells none
0.0
0.0
Dermal fibroblast IL-4
0.0
0.0

Dendritic cells LPS
0.0
0.0
Dermal Fibroblasts
0.0
0.0

rest

Dendritic cells anti-
0.0
0.0
Neutrophils
100.0
0.0

CD40

TNF a + LPS

Monocytes rest
0.0
0.0
Neutrophils rest
0.0
0.0

Monocytes LPS
0.0
43.2
Colon
0.0
0.0

Macrophages rest
0.0
0.0
Lung
0.0
100.0

Macrophages LPS
0.0
0.0
Thymus
0.0
0.0

HUVEC none
0.0
0.0
Kidney
0.0
0.0

HUVEC starved
0.0
0.0

[0884] Panel 1.3D Summary: Ag2299 The expression of the CG155769-01 gene appears to be highest in a sample derived from normal colon tissue (CT=33.9). In addition, there is substantial expression in testis tissue and a lung cancer cell line (SHP-77). Thus, the expression of this gene could be used to distinguish these samples from other samples in the panel. Moreover, therapeutic modulation of this gene, through the use of small molecule drugs, antibodies or protein therapeutics might be of benefit in the treatment of lung cancer.

[0885] Panel 2.2 Summary: Ag2299 The expression of the CG155769-01 gene is low/undetectable in all samples on this panel (CTs>35). (Data not shown.)

[0886] Panel 4.1D Summary: Ag2299 The expression of the CG155769-01 gene is exclusive to normal lung tissue. Thus, expression of this gene could potentially be used as a marker of lung tissue. This expression profile suggests that the protein encoded by this gene may be involved in the development and homeostasis of the lung. Therapeutic modulation of this gene product may be effective in maintaining and restoring normal function in the lung and may be useful in treating diseases that affect the lung.

[0887] AW. CG155882-01/GMAC055861_A: Olfactory Receptor

[0888] Expression of gene CG155882-01 was assessed using the primer-probe set Ag2192, described in Table AWA. Results of the RTQ-PCR runs are shown in Tables AWB, AWC, AWD and AWE.

223TABLE AWAProbe Name Ag2192StartSEQ IDPrimersSequencesLengthPositionNO:Forward5′-tgtattcatgggactcaccaa-3′2145421ProbeTET-5′-tcacgggagattcagcttctactttt-3′-TAMRA2667422Reverse5′-gctcgcaaagtagaacaacaaa-3′22102423

[0889]

224

TABLE AWB

Panel 1.3D

Rel. Exp. (%) Ag2192, Run

Rel. Exp. (%) Ag2192, Run

Tissue Name
165725843
Tissue Name
165725843

Liver adenocarcinoma
0.0
Kidney (fetal)
6.5

Pancreas
0.0
Renal ca. 786-0
21.5

Pancreatic ca. CAPAN 2
0.0
Renal ca. A498
27.9

Adrenal gland
0.0
Renal ca. RXF 393
32.8

Thyroid
0.0
Renal ca. ACHN
100.0

Salivary gland
0.0
Renal ca. UO-31
7.9

Pituitary gland
0.0
Renal ca. TK-10
20.2

Brain (fetal)
6.2
Liver
0.0

Brain (whole)
0.0
Liver (fetal)
0.0

Brain (amygdala)
0.0
Liver ca. (hepatoblast)
0.0

HepG2

Brain (cerebellum)
0.0
Lung
0.0

Brain (hippocampus)
0.0
Lung (fetal)
3.0

Brain (substantia nigra)
0.0
Lung ca. (small cell) LX-1
1.1

Brain (thalamus)
0.0
Lung ca. (small cell)
0.0

NCI-H69

Cerebral Cortex
0.0
Lung ca. (s.cell var.)
0.0

SHP-77

Spinal cord
0.0
Lung ca. (large cell)NCI-
0.0

H460

glio/astro U87-MG
15.0
Lung ca. (non-sm.cell)
0.0

A549

glio/astro U-118-MG
27.9
Lung ca. (non-s.cell)
0.0

NCI-H23

astrocytoma SW1783
14.1
Lung ca. (non-s.cell) 8.7

HOP-62

neuro*; met SK-N-AS
0.0
Lung ca. (non-s.cl) NCI-
2.9

H522

astrocytoma SF-539
0.0
Lung ca. (squam.) SW
14.8

900

astrocytoma SNB-75
0.0
Lung ca. (squam.) NCI-
0.0

H596

glioma SNB-19
0.0
Mammary gland
0.0

glioma U251
96.6
Breast ca.* (pl.ef) MCF-7
0.0

glioma SF-295
10.4
Breast ca.* (pl.ef) MDA-
0.0

MB-231

Heart (fetal)
0.0
Breast ca.* (pl.ef) T47D
0.0

Heart
0.0
Breast ca. BT-549
0.0

Skeletal muscle (fetal)
6.7
Breast ca. MDA-N
0.0

Skeletal muscle
0.0
Ovary
5.8

Bone marrow
7.2
Ovarian ca. OVCAR-3
0.0

Thymus
9.3
Ovarian ca. OVCAR-4
0.0

Spleen
0.0
Ovarian ca. OVCAR-5
0.0

Lymph node
0.0
Ovarian ca. OVCAR-8
0.0

Colorectal
1.8
Ovarian ca. IGROV-1
0.0

Stomach
0.0
Ovarian ca.* (ascites)
3.6

SK-OV-3

Small intestine
0.0
Uterus
0.0

Colon ca. SW480
0.0
Plancenta
0.0

Colon ca.* SW620(SW480
0.0
Prostate
3.2

met)

Colon ca. HT29
0.0
Prostate ca.* (bone
0.0

met)PC-3

Colon ca. HCT-116
3.3
Testis
0.0

Colon ca. CaCo-2
0.0
Melanoma Hs688(A).T
0.0

Colon ca.
0.0
Melanoma* (met)
0.0

tissue(ODO3866)

Hs688(B).T

Colon ca. HCC-2998
0.0
Melanoma UACC-62
37.9

Gastric ca.* (liver met)
0.0
Melanoma M14
45.1

NCI-N87

Bladder
0.0
Melanoma LOX IMVI
3.7

Trachea
0.0
Melanoma* (met) SK-
0.0

MEL-5

Kidney
0.0
Adipose
7.2

[0890]

225

TABLE AWC

Panel 2D

Rel. Exp. (%) Ag2192,

Rel. Exp. (%) Ag2192,

Tissue Name
Run 164024748
Tissue Name
Run 164024748

Normal Colon
0.0
Kidney Margin 8120608
0.0

CC Well to Mod Diff
12.9
Kidney Cancer 8120613
0.0

(ODO3866)

CC Margin (ODO3866)
12.3
Kidney Margin 8120614
7.2

CC Gr.2 rectosigmoid
0.0
Kidney Cancer 9010320
3.9

(ODO3868)

CC Margin (ODO3868)
0.0
Kidney Margin 9010321
20.6

CC Mod Diff (ODO3920)
0.0
Normal Uterus
0.0

CC Margin (ODO3920)
0.0
Uterus Cancer 064011
0.0

CC Gr.2 ascend colon
2.8
Normal Thyroid
0.0

(ODO3921)

CC Margin (ODO3921)
2.1
Thyroid Cancer 064010
0.0

CC from Partial Hepatectomy
0.0
Thyroid Cancer A302152
1.7

(ODO4309) Mets

Liver Margin (ODO4309)
0.0
Thyroid Margin A302153
2.5

Colon mets to lung (ODO4451-
0.0
Normal Breast
0.0

01)

Lung Margin (OD04451-02)
0.0
Breast Cancer (OD04566)
0.0

Normal Prostate 6546-1
0.0
Breast Cancer (OD04590-
0.0

01)

Prostate Cancer (OD04410)
0.0
Breast Cancer Mets
0.0

(OD04590-03)

Prostate Margin (OD04410)
0.0
Breast Cancer Metastasis
0.0

(OD04655-05)

Prostate Cancer (OD04720-01)
0.0
Breast Cancer 064006
0.0

Prostate Margin (OD04720-02)
5.0
Breast Cancer 1024
0.0

Normal Lung 061010
0.0
Breast Cancer 9100266
0.0

Lung Met to Muscle
26.4
Breast Margin 9100265
0.0

(ODO4286)

Muscle Margin (ODO4286)
0.0
Breast Cancer A209073
2.8

Lung Malignant Cancer
0.0
Breast Margin A2090734
0.0

(OD03126)

Lung Margin (OD03126)
0.0
Normal Liver
0.0

Lung Cancer (OD04404)
0.0
Liver Cancer 064003
0.0

Lung Margin (OD04404)
0.0
Liver Cancer 1025
2.0

Lung Cancer (OD04565)
0.0
Liver Cancer 1026
0.0

Lung Margin (OD04565)
2.9
Liver Cancer 6004-T
5.4

Lung Cancer (OD04237-01)
0.0
Liver Tissue 6004-N
3.2

Lung Margin (OD04237-02)
0.0
Liver Cancer 6005-T
0.0

Ocular Mel Met to Liver
4.9
Liver Tissue 6005-N
0.0

(ODO4310)

Liver Margin (ODO4310)
0.0
Normal Bladder
0.0

Melanoma Mets to Lung
2.5
Bladder Cancer 1023
0.0

(OD04321)

Lung Margin (OD04321)
0.0
Bladder Cancer A302173
9.2

Normal Kidney
25.0
Bladder Cancer
0.0

(OD04718-01)

Kidney Ca, Nuclear grade 2
100.0
Bladder Normal Adjacent
0.0

(OD04338)

(OD04718-03)

Kidney Margin (OD04338)
4.6
Normal Ovary
0.0

Kidney Ca Nuclear grade 1/2
48.0
Ovarian Cancer 064008
0.0

(OD04339)

Kidney Margin (OD04339)
18.9
Ovarian Cancer
0.0

(OD04768-07)

Kidney Ca, Clear cell type
26.8
Ovary Margin (OD04768-
0.0

(OD04340)

08)

Kidney Margin (OD04340)
12.0
Normal Stomach
0.0

Kidney Ca, Nuclear grade 3
7.0
Gastric Cancer 9060358
1.1

(OD04348)

Kidney Margin (OD04348)
2.5
Stomach Margin 9060359
0.0

Kidney Cancer (OD04622-01)
2.6
Gastric Cancer 9060395
2.1

Kidney Margin (OD04622-03)
5.6
Stomach Margin 9060394
2.1

Kidney Cancer (OD04450-01)
89.5
Gastric Cancer 9060397
0.0

Kidney Margin (OD04450-03)
7.2
Stomach Margin 9060396
0.0

Kidney Cancer 8120607
2.0
Gastric Cancer 064005
0.0

[0891]

226

TABLE AWD

Panel 3D

Rel. Exp. (%)

Rel. Exp. (%)

Ag2192, Run

Ag2192, Run

Tissue Name
164795770
Tissue Name
164795770

Daoy-Medulloblastoma
0.0
Ca Ski-Cervical epidermoid
1.2

carcinoma (metastasis)

TE671-Medulloblastoma
0.0
ES-2-Ovarian clear cell carcinoma
6.1

D283 Med-Medulloblastoma
0.0
Ramos-Stimulated with
0.0

PMA/ionomycin 6 h

PFSK-1-Primitive
0.0
Ramos-Stimulated with
0.0

Neuroectodermal

PMA/ionomycin 14 h

XF-498-CNS
64.6
MEG-01-Chronic myelogenous
0.0

leukemia (megokaryoblast)

SNB-78-Glioma
1.8
Raji-Burkitt's lymphoma
4.4

SF-268-Glioblastoma
0.0
Daudi-Burkitt's lymphoma
0.0

T98G-Glioblastoma
0.0
U266-B-cell plasmacytoma
0.0

SK-N-SH-Neuroblastoma
0.0
CA46-Burkitt's lymphoma
0.0

(metastasis)

SF-295-Glioblastoma
4.0
RL-non-Hodgkin's B-cell
0.0

lymphoma

Cerebellum
0.0
JM1-pre-B-cell lymphoma
0.0

Cerebellum
0.0
Jurkat-T cell leukemia
0.0

NCI-H292-Mucoepidermoid
0.0
TF-1-Erythroleukemia
0.0

lung carcinoma

DMS-114-Small cell lung
0.0
HUT 78-T-cell lymphoma
0.0

cancer

DMS-79-Small cell lung
0.0
U937-Histiocytic lymphoma
0.0

cancer

NCI-H146-Small cell lung
0.0
KU-812-Myelogenous leukemia
0.0

cancer

NCI-H526-Small cell lung
0.0
769-P-Clear cell renal carcinoma
44.1

cancer

NCI-N417-Small cell lung
0.0
Caki-2-Clear cell renal carcinoma
100.0

cancer

NCI-H82-Small cell lung
0.0
SW 839-Clear cell renal carcinoma
22.1

cancer

NCI-H157-Squamous cell
0.0
G401-Wilms' tumor
0.0

lung cancer (metastasis)

NCI-H1155-Large cell lung
0.0
Hs766T-Pancreatic carcinoma (LN
0.0

cancer

metastasis)

NCI-H1299-Large cell lung
1.1
CAPAN-1-Pancreatic
0.0

cancer

adenocarcinoma (liver metastasis)

NCI-H727-Lung carcinoid
0.0
SU86.86-Pancreatic carcinoma
0.0

(liver metastasis)

NCI-UMC-11-Lung
0.0
BxPC-3-Pancreatic
0.0

carcinoid

adenocarcinoma

LX-1-Small cell lung cancer
0.0
HPAC-Pancreatic adenocarcinoma
0.0

Colo-205-Colon cancer
0.0
MIA PaCa-2-Pancreatic carcinoma
0.0

KM12-Colon cancer
0.0
CFPAC-1-Pancreatic ductal
0.0

adenocarcinoma

KM20L2-Colon cancer
0.0
PANC-1-Pancreatic epithelioid
0.0

ductal carcinoma

NCI-H716-Colon cancer
0.0
T24-Bladder carcinma (transitional
2.0

cell)

SW-48-Colon
0.0
5637-Bladder carcinoma
0.0

adenocarcinoma

SW1116-Colon
0.0
HT-1197-Bladder carcinoma
0.0

adenocarcinoma

LS 174T-Colon
0.0
UM-UC-3-Bladder carcinma
0.0

adenocarcinoma

(transitional cell)

SW-948-Colon
0.0
A204-Rhabdomyosarcoma
0.0

adenocarcinoma

SW-480-Colon
0.0
HT-1080-Fibrosarcoma
29.1

adenocarcinoma

NCI-SNU-5-Gastric
0.0
MG-63-Osteosarcoma
0.0

carcinoma

KATO III-Gastric carcinoma
0.0
SK-LMS-1-Leiomyosarcoma
23.7

(vulva)

NCI-SNU-16-Gastric
7.5
SJRH30-Rhabdomyosarcoma (met
2.2

carcinoma

to bone marrow)

NCI-SNU-1-Gastric
0.0
A431-Epidermoid carcinoma
0.0

carcinoma

RF-1-Gastric
0.0
WM266-4-Melanoma
22.5

adenocarcinoma

RF-48-Gastric
0.0
DU 145-Prostate carcinoma (brain
0.0

adenocarcinoma

metastasis)

MKN-45-Gastric carcinoma
0.0
MDA-MB-468-Breast
0.0

adenocarcinoma

NCI-N87-Gastric carcinoma
0.0
SCC-4-Squamous cell carcinoma
0.0

of tongue

OVCAR-5-Ovarian
0.0
SCC-9-Squamous cell carcinoma
0.0

carcinoma

of tongue

RL95-2-Uterine carcinoma
0.0
SCC-15-Squamous cell carcinoma
0.0

of tongue

HelaS3-Cervical
0.0
CAL 27-Squamous cell carcinoma
0.0

adenocarcinoma

of tongue

[0892]

227

TABLE AWE

Panel 4D

Rel. Exp. (%) Ag2192,

Rel. Exp. (%) Ag2192,

Tissue Name
Run 163588121
Tissue Name
Run 163588121

Secondary Th1 act
0.0
HUVEC IL-1beta
0.0

Secondary Th2 act
0.0
HUVEC IFN gamma
0.0

Secondary Tr1 act
12.3
HUVEC TNF alpha + IFN
0.0

gamma

Secondary Th1 rest
0.0
HUVEC TNF alpha + IL4
0.0

Secondary Th2 rest
0.0
HUVEC IL-11
0.0

Secondary Tr1 rest
0.0
Lung Microvascular EC none
0.0

Primary Th1 act
0.0
Lung Microvascular EC
24.3

TNFalpha + IL-1beta

Primary Th2 act
0.0
Microvascular Dermal EC none
0.0

Primary Tr1 act
0.0
Microsvasular Dermal EC
0.0

TNFalpha + IL-1beta

Primary Th1 rest
0.0
Bronchial epithelium TNFalpha +
0.0

IL1beta

Primary Th2 rest
0.0
Small airway epithelium none
0.0

Primary Tr1 rest
0.0
Small airway epithelium
0.0

TNFalpha + IL-1beta

CD45RA CD4 lymphocyte
0.0
Coronery artery SMC rest
0.0

act

CD45RO CD4 lymphocyte
0.0
Coronery artery SMC TNFalpha +
0.0

act

IL-1beta

CD8 lymphocyte act
0.0
Astrocytes rest
0.0

Secondary CD8
0.0
Astrocytes TNFalpha + IL-1beta
0.0

lymphocyte rest

Secondary CD8
0.0
KU-812 (Basophil) rest
0.0

lymphocyte act

CD4 lymphocyte none
0.0
KU-812 (Basophil)
0.0

PMA/ionomycin

2ry Th1/Th2/Tr1_anti-
0.0
CCD1106 (Keratinocytes) none
0.0

CD95 CH11

LAK cells rest
0.0
CCD1106 (Keratinocytes)
0.0

TNFalpha + IL-1beta

LAK cells IL-2
0.0
Liver cirrhosis
38.7

LAK cells IL-2 + IL-12
0.0
Lupus kidney
16.7

LAK cells IL-2 + IFN
0.0
NCI-H292 none
0.0

gamma

LAK cells IL-2 + IL-18
0.0
NCI-H292 IL-4
0.0

LAK cells
0.0
NCI-H292 IL-9
0.0

PMA/ionomycin

NK Cells IL-2 rest
0.0
NCI-H292 IL-13
0.0

Two Way MLR 3 day
0.0
NCI-H292 IFN gamma
0.0

Two Way MLR 5 day
0.0
HPAEC none
0.0

Two Way MLR 7 day
0.0
HPAEC TNF alpha + IL-1 beta
0.0

PBMC rest
0.0
Lung fibroblast none
67.8

PBMC PWM
0.0
Lung fibroblast TNF alpha + IL-
0.0

1 beta

PBMC PHA-L
20.0
Lung fibroblast IL-4
76.3

Ramos (B cell) none
0.0
Lung fibroblast IL-9
65.1

Ramos (B cell) ionomycin
0.0
Lung fibroblast IL-13
66.0

B lymphocytes PWM
20.9
Lung fibroblast IFN gamma
0.0

B lymphocytes CD40L
0.0
Dermal fibroblast CCD1070 rest
0.0

and IL-4

EOL-1 dbcAMP
0.0
Dermal fibroblast CCD1070
22.2

TNF alpha

EOL-1 dbcAMP
0.0
Dermal fibroblast CCD1070 IL-
19.9

PMA/ionomycin

1 beta

Dendritic cells none
0.0
Dermal fibroblast IFN gamma
0.0

Dendritic cells LPS
0.0
Dermal fibroblast IL-4
0.0

Dendritic cells anti-CD40
0.0
IBD Colitis 2
18.2

Monocytes rest
0.0
IBD Crohn's
0.0

Monocytes LPS
0.0
Colon
40.6

Macrophages rest
0.0
Lung
0.0

Macrophages LPS
0.0
Thymus
100.0

HUVEC none
0.0
Kidney
0.0

HUVEC starved
0.0

[0893] Panel 1.3D Summary: Ag2192 The expression of the CG155882-01 gene is highest in a sample derived from a renal cancer cell line (ACHN) (CT=33.3). In addition, there is expression in another renal cancer cell line, two melanoma cell lines and a glioma cell line. Thus the expression of this gene could be used to distinguish these samples from others in the panel. Moreover, therapeutic modulation of this gene, through the use of small molecule drugs, antibodies or protein therapeutics might be of benefit in the treatment of renal cancer, melanoma or glioma.

[0894] Panel 2D Summary: Ag2192 The expression of the CG155882-01 gene appears to be highest in a sample derived from a kidney cancer(CT=33.3). In addition, there appears to be substantial expression in several other kidney cancer samples. This result is in corcordance with the result seen in Panel 1.3D. Of note is the difference in expression between kidney cancer samples and their respective normal adjacent tissues. Thus, the expression of this gene could be used to distinguish kidney cancer samples from the rest of the samples in the panel. Moreover, therapeutic modulation of this gene, through the use of small molecule drugs, antibodies or protein therapeutics might be of benefit in the treatment of kidney cancer.

[0895] Panel 3D Summary: Ag2192 The expression of the CG155882-01 gene appears to be highest in samples derived from kidney cancer cell lines (CT=32.6). This association with kidney cancer is also seen in Panels 1.3D and 2D. In addition, there is substantial expression seen in one brain cancer cell line, one fibrosarcoma cell line, one melanoma cell line and one leiomyosarcoma cell line. Thus, the expression of this gene could be used to distinguish these samples from other samples in the panel. Moreover, therapeutic modulation of this gene, through the use of small molecule drugs, antibodies or protein therapeutics might be of benefit in the treatment of kidney cancer, melanoma, fiborsarcoma, brain cancer, or leiomyosarcoma.

[0896] Panel 4D Summary: Ag 2192 The expression of the CG155882-01 gene is higher in untreated fibroblasts than in fibroblasts treated by the potent inflammatory cytokines TNF-a and IFN-g cytokines but not by IL-4 cytokine. IL-4 has been associated with anti-inflammatory properties. TNF-a and IFNg have been shown to lead to the activation of proteolytic degradation of extracellular matrix in fibroblasts, a phenomenon associated with emphysema. IFN g has also been shown to lead to direct granulomatous inflammation of the lung. Therefore, therapeutic modulation of this gene product, through the use of small molecule drugs, or antibodies might be beneficial for the treatment of these diseases.

[0897] AX. CG55766-02/GMAC044810_B: Olfactory Receptor

[0898] Expression of gene CG55766-02 was assessed using the primer-probe set Ag2182, described in Table AXA. Results of the RTQ-PCR runs are shown in Tables AXB and AXC.

228TABLE AXAProbe Name Ag2182StartSEQ IDPrimersSequencesLengthPositionNO:Forward5′-aagctgtgtggttgaattcatc-3′2287424ProbeTET-5′-tctaactatcctgagctccaggggca-3′-TAMRA26121425Reverse5′-taaataaccaggaaagccacaa-3′22152426

[0899]

229

TABLE AXB

Panel 2D

Rel. Exp. (%) Ag2182,

Rel. Exp. (%) Ag2182,

Tissue Name
Run 163582696
Tissue Name
Run 163582696

Normal Colon
0.0
Kidney Margin 8120608
0.0

CC Well to Mod Diff
0.0
Kidney Cancer 8120613
0.0

(ODO3866)

CC Margin (ODO3866)
9.1
Kidney Margin 8120614
0.0

CC Gr.2 rectosigmoid
0.0
Kidney Cancer 9010320
0.0

(ODO3868)

CC Margin (ODO3868)
0.0
Kidney Margin 9010321
0.0

CC Mod Diff (ODO3920)
0.0
Normal Uterus
0.0

CC Margin (ODO3920)
0.0
Uterus Cancer 064011
0.0

CC Gr.2 ascend colon
0.0
Normal Thyroid
0.0

(ODO3921)

CC Margin (ODO3921)
21.5
Thyroid Cancer 064010
0.0

CC from Partial Hepatectomy
0.0
Thyroid Cancer A302152
0.0

(ODO4309) Mets

Liver Margin (ODO4309)
0.0
Thyroid Margin A302153
0.0

Colon mets to lung (OD04451-
0.0
Normal Breast
63.7

01)

Lung Margin (OD04451-02)
0.0
Breast Cancer (OD04566)
0.0

Normal Prostate 6546-1
0.0
Breast Cancer (OD04590-
0.0

01)

Prostate Cancer (OD04410)
11.3
Breast Cancer Mets
0.0

(OD04590-03)

Prostate Margin (OD04410)
21.0
Breast Cancer Metastasis
0.0

(OD04655-05)

Prostate Cancer (OD04720-01)
94.6
Breast Cancer 064006
8.2

Prostate Margin (OD04720-02)
100.0
Breast Cancer 1024
34.4

Normal Lung 061010
16.2
Breast Cancer 9100266
0.0

Lung Met to Muscle
14.1
Breast Margin 9100265
0.0

(ODO4286)

Muscle Margin (ODO4286)
0.0
Breast Cancer A209073
11.8

Lung Malignant Cancer
0.0
Breast Margin A2090734
47.6

(OD03126)

Lung Margin (OD03126)
27.9
Normal Liver
0.0

Lung Cancer (OD04404)
0.0
Liver Cancer 604003
1.5

Lung Margin (OD04404)
0.0
Liver Cancer 1025
0.0

Lung Cancer (OD04565)
0.0
Liver Cancer 1026
0.0

Lung Margin (OD04565)
33.0
Liver Cancer 6004-T
0.0

Lung Cancer (OD04237-01)
0.0
Liver Tissue 6004-N
0.0

Lung Margin (OD04237-02)
0.0
Liver Cancer 6005-T
0.0

Ocular Mel Met to Liver
0.0
Liver Tissue 6005-N
0.0

(ODO4310)

Liver Margin (ODO4310)
0.0
Normal Bladder
13.6

Melanoma Mets to Lung
0.0
Bladder Cancer 1023
4.8

(OD04321)

Lung Margin (OD04321)
34.2
Bladder Cancer A302173
31.6

Normal Kidney
17.2
Bladder Cancer
0.0

(OD04718-01)

Kidney Ca, Nuclear grade 2
0.0
Bladder Normal Adjacent
0.0

(OD04338)

(OD04718-03)

Kidney Margin (OD04338)
23.0
Normal Ovary
0.0

Kidney Ca Nuclear grade 1/2
0.0
Ovarian Cancer 064008
15.3

(OD04339)

Kidney Margin (OD04339)
0.0
Ovarian Cancer
14.5

(OD04768-07)

Kidney Ca, Clear cell type
0.0
Ovary Margin (OD04768-
0.0

(OD04340)

08)

Kidney Margin (OD04340)
32.1
Normal Stomach
4.2

Kidney Ca, Nuclear grade 3
4.5
Gastric Cancer 9060358
0.0

(OD04348)

Kidney Margin (OD04348)
57.0
Stomach Margin 9060359
0.0

Kidney Cancer (OD04622-01)
0.0
Gastric Cancer 9060395
0.0

Kidney Margin (OD04622-03)
0.0
Stomach Margin 9060394
3.7

Kidney Cancer (OD04450-01)
0.0
Gastric Cancer 9060397
0.0

Kidney Margin (OD04450-03)
0.0
Stomach Margin 9060396
0.0

Kidney Cancer 8120607
0.0
Gastric Cancer 064005
7.9

[0900]

230

TABLE AXC

Panel 4D

Rel. Exp. (%) Ag2182,

Rel. Exp. (%) Ag2182,

Tissue Name
Run 163578421
Tissue Name
Run 163578421

Secondary Th1 act
0.0
HUVEC IL-1beta
0.0

Secondary Th2 act
0.0
HUVEC IFN gamma
1.3

Secondary Tr1 act
0.0
HUVEC TNF alpha + IFN
0.0

gamma

Secondary Th1 rest
0.0
HUVEC TNF alpha + IL4
0.0

Secondary Th2 rest
0.0
HUVEC IL-11
0.0

Secondary Tr1 rest
0.0
Lung Microvascular EC none
0.0

Primary Th1 act
0.0
Lung Microvascular EC
0.0

TNFalpha + IL-1beta

Primary Th2 act
0.0
Microvascular Dermal EC none
0.0

Primary Tr1 act
0.0
Microsvasular Dermal EC
0.0

TNFalpha + IL-1beta

Primary Th1 rest
0.0
Bronchial epithelium TNFalpha +
2.5

IL1beta

Primary Th2 rest
0.0
Small airway epithelium none
3.7

Primary Tr1 rest
0.0
Small airway epithelium
44.1

TNFalpha + IL-1beta

CD45RA CD4 lymphocyte
0.0
Coronery artery SMC rest
0.0

act

CD45RO CD4 lymphocyte
0.0
Coronery artery SMC TNFalpha +
0.0

act

IL-1beta

CD8 lymphocyte act
0.0
Astrocytes rest
1.7

Secondary CD8
0.0
Astrocytes TNFalpha + IL-1beta
3.0

lymphocyte rest

Secondary CD8
0.0
KU-812 (Basophil) rest
0.0

lymphocyte act

CD4 lymphocyte none
0.0
KU-812 (Basophil)
0.0

PMA/ionomycin

2ry Th1/Th2/Tr_anti-
0.0
CCD1106 (Keratinocytes) none
42

CD95 CH11

LAK cells rest
0.0
CCD1106 (Keratinocytes)
1.8

TNFalpha + IL-1beta

LAK cells IL-2
0.0
Liver cirrhosis
6.0

LAK cells IL-2 + IL-12
0.0
Lupus kidney
0.8

LAK cells IL-2 + IFN
0.0
NCI-H292 none
77.4

gamma

LAK cells IL-2 + IL-18
0.0
NCI-H292 IL-4
100.0

LAK cells
0.0
NCI-H292 IL-9
62.4

PMA/ionomycin

NK Cells IL-2 rest
0.0
NCI-H292 IL-13
35.4

Two Way MLR 3 day
0.0
NCI-H292 IFN gamma
22.4

Two Way MLR 5 day
0.0
HPAEC none
0.0

Two Way MLR 7 day
0.0
HPAEC TNF alpha + IL-1 beta
0.0

PBMC rest
0.0
Lung fibroblast none
3.1

PBMC PWM
0.0
Lung fibroblast TNF alpha + IL-
0.9

1 beta

PBMC PHA-L
0.0
Lung fibroblast IL-4
1.7

Ramos (B cell) none
2.6
Lung fibroblast IL-9
3.0

Ramos (B cell) ionomycin
0.0
Lung fibroblast IL-13
1.4

B lymphocytes PWM
0.0
Lung fibroblast IFN gamma
0.8

B lymphocytes CD40L
0.0
Dermal fibroblast CCD1070 rest
0.6

and IL-4

EOL-1 dbAMP
0.0
Dermal fibroblast CCD1070
0.0

TNF alpha

EOL-1 dbcAMP
0.0
Dermal fibroblast CCD1070 IL-
0.0

PMA/ionomycin

1 beta

Dendritic cells none
0.0
Dermal fibroblast IFN gamma
0.0

Dendritic cells LPS
0.0
Dermal fibroblast IL-4
0.0

Dendritic cells anti-CD40
0.0
IBD Colitis 2
1.8

Monocytes rest
0.0
IBD Crohn's
0.0

Monocytes LPS
0.0
Colon
1.4

Macrophages rest
0.0
Lung
1.9

Macrophages LPS
0.0
Thymus
0.6

HUVEC none
0.0
Kidney
0.0

HUVEC starved
1.4

[0901] CNS_neurodegeneration_v1.0 Summary: Ag2182 Expression of the CG55766-02 gene is low/undetectable in all samples in this panel (CTs>35). (Data not shown.)

[0902] Panel 1.3D Summary: Ag2182 Expression of the CG55766-02 gene is low/undetectable in all samples in this panel (CTs>35). (Data not shown.)

[0903] Panel 2.2 Summary: Ag2182 Expression of the CG55766-02 gene is low/undetectable in all samples in this panel (CTs>35). (Data not shown.)

[0904] Panel 2D Summary: Ag2182 The expression of the CG55766-02 gene appears to be highest in normal prostate (CT=33.5). In addition, there appears to be substantial expression in prostate cancer adjacent to normal prostate and in normal breast and kidney tissue. Of note was the differential expression between many of the normal tissues when compared to their malignant counterparts. Thus, expression of this gene could be used to distinguish between these samples and other samples on this panel and in particular distinguish between normal and cancer. Moreover, therapeutic modulation of this gene, through the use of small molecule drugs, antibodies or protein therapeutics might be of benefit for the treatment of breast cancer, kidney cancer or prostate cancer.

[0905] Panel 3D Summary: Ag2182 Expression of the CG55766-02 gene is low/undetectable in all samples in this panel (CTs>35). (Data not shown.)

[0906] Panel 4D Summary: Ag2182 The CG55766-02 gene is expressed at a moderate level (CT=28=31) in resting and IL-4, IL-9, or IL-13 and IFN gamma activated NCI-H292 mucoepidermoid cells. Moderate expression of this genee is also detected in the TNFalpha+IL-1beta stimulated small airway epithelial cells, while low but significant levels of expression (CT 33-35) are detected in IL-9 treated lung fibroblasts, untreated lung fibroblasts, TNFalpha+IL-1beta treated bronchial epithelial cells, and untreated Ramos B cells. The expression of this gene in lung derived cells and B cells suggests that this gene may be involved in normal conditions as well as pathological and inflammatory lung disorders including chronic obstructive pulmonary disease, asthma, allergy and emphysema. Therefore, small molecules or antibodies that modulate the function of this gene product may be useful therapeutics for the reduction or elimination of the symptoms in these diseases.

[0907] AY. CG156086-01/GMbA144L1_C: Olfactory Receptor

[0908] Expression of gene CG156086-01 was assessed using the primer-probe sets Ag1369, Ag1398, Ag1401, Ag1650, Ag2434 and Ag1618, described in Tables AYA, AYB, AYC, AYD, AYE, and AYF. Results of the RTQ-PCR runs are shown in Tables AYG and AYH.

231TABLE AYAProbe Name Ag1398StartSEQ IDPrimersSequencesLengthPositionNO:Forward5′-actcctcaaccccatcatttac-3′22868427ProbeTET-5′-accatgaagtaaaggcagccctcaga-3′-TAMRA26900428Reverse5′-cacttcctctgcaatgtatggt-3′22929429

[0909]

232

TABLE AYB

Probe Name Ag1650

Start
SEQ ID

Primers
Sequences
Length
Position
NO:

Forward
5′-actcctcaaccccatcatttac-3′
22
868
430

Probe
TET-5′-accatgaagtaaaggcagccctcaga-3′-TAMRA
26
900
431

Reverse
5′-cacttcctctgcaatgtatggt-3′
22
929
432

[0910]

233

TABLE AYC

Probe Name Ag2434

Start
SEQ ID

Primers
Sequences
Length
Position
NO:

Forward
5′-agtcatcagccccaagatg-3′
19
244
433

Probe
TET-5′-tgttgacttcctcagtcatgacaaga-3′-TAMRA
26
265
434

Reverse
5′-ttgagtcatgcagccattg-3′
19
301
435

[0911]

234

TABLE AYD

Probe Name Ag1618

Start
SEQ ID

Primers
Sequences
Length
Position
NO:

Forward
5′-actcctcaaccccatcatttac-3′
22
868
436

Probe
TET-5′-accatgaagtaaaggcagccctcaga-3′-TAMRA
26
900
437

Reverse
5′-cacttcctctgcaatgtatggt-3′
22
929
438

[0912]

235

TABLE AYE

Probe Name Ag1369

Start
SEQ ID

Primers
Sequences
Length
Position
NO:

Forward
5′-actcctcaaccccatcatttac-3′
22
868
439

Probe
TET-5′-accatgaagtaaaggcagccctcaga-3′-TAMRA
26
900
440

Reverse
5′-cacttcctctgcaatgtggt-3′
22
929
441

[0913]

236

TABLE AYF

Probe Name Ag1401

Start
SEQ ID

Primers
Sequences
Length
Position
NO:

Forward
5′-actcctcaaccccatcatttac-3′
22
868
442

Probe
TET-5′-accatgaagtaaaggcagccctcaga-3′-TAMRA
26
900
443

Reverse
5′-cacttcctctgcaatgtggt-3′
22
929
444

[0914]

237

TABLE AYG

Panel 1.2

Rel. Exp. (%)
Rel. Exp. (%)

Rel. Exp. (%)
Rel. Exp. (%)

Ag1398, Run
Ag1398, Run

Ag1398, Run
Ag1398, Run

Tissue Name
138382712
139735391
Tissue Name
138382712
139735391

Endothelial cells
0.0
0.0
Renal ca. 786-0
0.0
0.0

Heart (Fetal)
0.0
0.0
Renal ca. A498
0.0
0.0

Pancreas
0.0
0.0
Renal ca. RXF
0.0
0.0

393

Pancreatic ca.
0.0
0.0
Renal ca. ACHN
0.0
0.2

CAPAN 2

Adrenal Gland
0.0
0.0
Renal ca. UO-31
0.0
1.1

Thyroid
0.0
0.0
Renal ca. TK-10
0.0
0.0

Salivary gland
0.0
0.0
Liver
0.0
0.0

Pituitary gland
0.0
0.0
Liver (fetal)
0.0
0.0

Brain (fetal)
0.0
0.0
Liver ca.
0.0
0.0

(hepatoblast)

HepG2

Brain (whole)
0.0
0.0
Lung
0.0
0.0

Brain (amygdala)
0.0
0.0
Lung (fetal)
0.0
0.0

Brain (cerebellum)
0.0
0.0
Lung ca. (small
0.0
0.0

cell) LX-1

Brain
0.0
0.0
Lung ca. (small
36.1
45.4

(hippocampus)

cell) NCI-H69

Brain (thalamus)
0.0
0.0
Lung ca. (s.cell
0.0
0.0

var.) SHP-77

Cerebral Cortex
0.0
0.0
Lung ca. (large
0.4
18.7

cell)NCI-H460

Spinal cord
0.0
0.0
Lung ca. (non-
14.2
0.4

sm. cell) A549

glio/astro U87-MG
0.0
0.0
Lung ca. (non-
0.0
0.0

s.cell) NCI-H23

glio/astro U-118-
0.0
0.0
Lung ca. (non-
0.0
0.0

MG

s.cell) HOP-62

astrocytoma
0.0
0.0
Lung ca. (non-
0.0
0.0

SW1783

s.cl) NCI-H522

neuro*; met SK-N-
0.0
0.0
Lung ca.
0.0
0.0

AS

(squam.) SW 900

astrocytoma SF-
0.0
0.0
Lung ca.
0.3
20.0

539

(squam.) NCI-

H596

astrocytoma SNB-
0.0
0.0
Mammary gland
0.0
0.0

75

glioma SNB-19
26.4
26.6
Breast ca.* (pl.ef)
0.0
0.0

MCF-7

glioma U251
0.0
0.0
Breast ca.* (pl.ef)
0.0
0.0

MDA-MB-231

glioma SF-295
0.0
0.0
Breast ca.* (pl.
16.6
44.8

ef) T47D

Heart
0.0
5.1
Breast ca. BT-
0.0
0.0

549

Skeletal Muscle
0.0
0.0
Breast ca. MDA-N
0.0
2.0

Bone marrow
0.0
0.0
Ovary
0.0
0.0

Thymus
0.0
0.0
Ovarian ca.
0.0
0.0

OVCAR-3

Spleen
0.0
0.0
Ovarian ca.
0.0
0.0

OVCAR-4

Lymph node
0.0
0.0
Ovarian ca.
100.0
100.0

OVCAR-5

Colorectal Tissue
0.0
0.4
Ovarian ca.
0.0
0.5

OVCAR-8

Stomach
0.0
0.0
Ovarian ca.
0.2
0.0

IGROV-1

Small intestine
0.0
0.0
Ovarian ca.
0.3
0.0

(ascites) SK-OV-3

Colon ca. SW480
0.0
0.0
Uterus
0.0
0.0

Colon ca.* SW620
0.0
0.0
Placenta
0.0
0.0

(SW480 met)

Colon ca. HT29
1.2
0.2
Prostate
0.0
0.0

Colon ca. HCT-
0.0
0.0
Prostate ca.*
0.0
0.0

116

(bone met) PC-3

Colon ca. CaCo-2
0.0
0.0
Testis
0.0
0.0

Colon ca. Tissue
1.0
11.9
Melanoma
0.0
0.0

(ODO3866)

Hs688(A).T

Colon ca. HCC-
0.0
0.0
Melanoma* (met)
0.0
1.0

2998

Hs688(B).T

Gastric ca.* (liver
0.0
0.0
Melanoma
0.0
0.0

met) NCI-N87

UACC-62

Bladder
0.0
0.0
Melanoma M14
0.0
0.0

Trachea
0.0
0.0
Melanoma LOX
0.0
0.0

IMVI

Kidney
0.0
0.0
Melanoma* (met)
0.0
0.0

SK-MEL-5

Kidney (fetal)
69.7
0.0

[0915]

238

TABLE AYH

Panel 4D

Rel. Exp. (%)
Rel. Exp. (%)

Rel. Exp. (%)
Rel. Exp. (%)

Ag1618, Run
Ag2434, Run

Ag1618, Run
Ag2434, Run

Tissue Name
165425781
164183931
Tissue Name
165425781
164183931

Secondary Th1 act
0.0
0.0
HUVEC IL-1beta
0.0
0.0

Secondary Th2 act
0.0
0.0
HUVEC IFN gamma
0.0
0.0

Secondary Tr1 act
9.4
0.0
HUVEC TNF alpha +
0.0
0.0

IFN gamma

Secondary Th1 rest
0.0
0.0
HUVEC TNF alpha +
0.0
0.0

IL4

Secondary Th2 rest
0.0
0.0
HUVEC IL-11
0.0
8.2

Secondary Tr1 rest
0.0
0.0
Lung Microvascular
0.0
0.0

EC none

Primary Th1 act
0.0
0.0
Lung Microvascular
0.0
0.0

EC TNF alpha + IL-

1beta

Primary Th2 act
0.0
0.0
Microvascular
0.0
0.0

Dermal EC none

Primary Tr1 act
0.0
0.0
Microsvasular Dermal
0.0
0.0

EC TNF alpha + IL-

1beta

Primary Th1 rest
17.4
0.0
Bronchial epithelium
0.0
0.0

TNF alpha + IL1beta

Primary Th2 rest
0.0
0.0
Small airway
0.0
0.0

epithelium none

Primary Tr1 rest
0.0
0.0
Small airway
0.0
0.0

epithelium TNF

alpha + IL-1beta

CD45RA CD4
0.0
0.0
Coronery artery SMC
0.0
0.0

lymphocyte act

rest

CD45RO CD4
0.0
0.0
Coronery artery SMC
0.0
0.0

lymphocyte act

TNF alpha + IL-1beta

CD8 lymphocyte act
0.0
35.8
Astrocytes rest
0.0
0.0

Secondary CD8
0.0
0.0
Astrocytes TNF
0.0
0.0

lymphocyte rest

alpha + IL-1beta

Secondary CD8
0.0
0.0
KU-812 (Basophil)
0.0
0.0

lymphocyte act

rest

CD4 lymphocyte
0.0
0.0
KU-812 (Basophil)
0.0
0.0

none

PMA/ionomycin

2ry
0.0
0.0
CCD1106
0.0
0.0

Th1/Th2/Tr1_anti-

(Keratinocytes) none

CD95 CH11

LAK cells rest
0.0
0.0
CCD1106
0.0
0.0

(Keratinocytes)

TNF alpha + IL-1beta

LAK cells IL-2
0.0
0.0
Liver cirrhosis
100.0
100.0

LAK cells IL-2 + IL-
0.0
0.0
Lupus kidney
0.0
0.0

12

LAK cells IL-2 + IFN
0.0
0.0
NCI-H292 none
0.0
0.0

gamma

LAK cells IL-2 + IL-
0.0
0.0
NCI-H292 IL-4
0.0
0.0

18

LAK cells
0.0
0.0
NCI-H292 IL-9
0.0
0.0

PMA/ionomycin

NK Cells IL-2 rest
0.0
0.0
NCI-H292 IL-13
0.0
0.0

Two Way MLR 3
0.0
0.0
NCI-H292 IFN
0.0
0.0

day

gamma

Two Way MLR 5
0.0
0.0
HPAEC none
0.0
0.0

day

Two Way MLR 7
0.0
0.0
HPAEC TNF alpha +
0.0
0.0

day

IL-1beta

PBMC rest
0.0
0.0
Lung fibroblast none
0.0
0.0

PBMC PWM
0.0
0.0
Lung fibroblast TNF
0.0
0.0

alpha + IL-1beta

PBMC PHA-L
0.0
0.0
Lung fibroblast IL-4
0.0
0.0

Ramos (B cell) none
0.0
0.0
Lung fibroblast IL-9
0.0
0.0

Ramos (B cell)
0.0
0.0
Lung fibroblast IL-13
0.0
0.0

ionomycin

B lymphocytes PWM
0.0
0.0
Lung fibroblast IFN
0.0
0.0

gamma

B lymphocytes
0.0
0.0
Dermal fibroblast
0.0
0.0

CD40L and IL-4

CCD1070 rest

EOL-1 dbcAMP
20.7
0.0
Dermal fibroblast
0.0
0.0

CCD1070 TNF alpha

EOL-1 dbcAMP
0.0
0.0
Dermal fibroblast
0.0
0.0

PMA/ionomycin

CCD1070 IL-1beta

Dendritic cells none
0.0
0.0
Dermal fibroblast IFN
0.0
0.0

gamma

Dendritic cells LPS
0.0
0.0
Dermal fibroblast IL-4
0.0
0.0

Dendritic cells anti-
0.0
0.0
IBD Colitis 2
2.4
0.0

CD40

Monocytes rest
0.0
0.0
IBD Crohn's
0.0
0.0

Monocytes LPS
13.6
0.0
Colon
0.0
0.0

Macrophages rest
0.0
0.0
Lung
0.0
0.0

Macrophages LPS
0.0
0.0
Thymus
0.0
0.0

HUVEC none
0.0
0.0
Kidney
19.2
30.1

HUVEC starved
0.0
39.5

[0916] Panel 1.2 Summary: Ag1398 Results from two experiments using this probe/primer set are in reasonable agreement. Expression of the CG156086-01 gene is limited to a single breast cancer cell line, a single ovarian cancer cell line, a single lung cancer cell line, and a single CNS cancer cell line. Thus, the therapeutic inhibition of CG156086-01 gene activity, through the use of small molecule drugs or antibodies, might be of utility in the treatment of the above listed cancer types. Ag1369/Ag1401 Expression of the CG156086-01 gene is low/undetectable (CT values>35) in all samples. (Data not shown.)

[0917] Panel 1.3D Summary: Ag1650/Ag2434 Expression of the CG156086-01 gene is low/undetected in all samples in this panel (CT>35). (Data not shown.)

[0918] Panel 2.2 Summary: Ag1650/Ag2434 Expression of the CG156086-01 gene is low/undetected in all samples in this panel (CT>35). (Data not shown.)

[0919] Panel 4D Summary: Ag1618/Ag2434 Results from two experiments using different probe/primer sets are in reasonable agreement. The CG156086-01 transcript is only detected in liver cirrhosis. Furthermore, this transcript is not detected in normal liver in Panel 1.3D, suggesting that CG156086-01 gene expression is unique to liver cirrhosis. The CG156086-01 gene encodes a putative GPCR; therefore, antibodies or small molecule therapeutics could reduce or inhibit fibrosis that occurs in liver cirrhosis. In addition, antibodies to this putative GPCR could also be used for the diagnosis of liver cirrhosis. Ag1650 Expression of the CG156086-01 gene is low/undetectable (CT values>35) across the samples on this panel.

[0920] AZ. CG50271-01: Olfactory Receptor

[0921] Expression of gene CG50271-01 was assessed using the primer-probe sets Ag2507 and Ag716, described in Tables AZA and AZB. Results of the RTQ-PCR runs are shown in Table AZC.

239TABLE AZAProbe Name Ag2507StartSEQ IDPrimersSequencesLengthPositionNO:Forward5′-tgtatttcttcctgagccagtt-3′22151445ProbeTET-5′-tgttatccatccactaccatacccaa-3′-TAMRA26189446Reverse5′-cacaaaagtgtcagcaatcatc-3′22215447

[0922]

240

TABLE AZB

Probe Name Ag1716

Start
SEQ ID

Primers
Sequences
Length
Position
NO:

Forward
5′-agagatgcagcatacatgcttt-3′
22
38
448

Probe
TET-5′-ctctttcatgtgctcactgtcctggg-3′-TAMRA
26
72
449

Reverse
5′-tctagcattgatggtgatgatg-3′
22
110
450

[0923]

241

TABLE AZC

Panel 1.3D

Rel. Exp. (%) Ag2507, Run

Rel. Exp. (%) Ag2507, Run

Tissue Name
165531073
Tissue Name
165531073

Liver adenocarcinoma
0.0
Kidney (fetal)
0.0

Pancreas
0.0
Renal ca. 786-0
0.0

Pancreatic ca. CAPAN 2
0.0
Renal ca. A498
0.0

Adrenal gland
0.0
Renal ca. RXF 393
0.0

Thyroid
0.0
Renal ca. ACHN
0.0

Salivary gland
0.0
Renal ca. UO-31
0.0

Pituitary gland
0.0
Renal ca. TK-10
0.0

Brain (fetal)
0.0
Liver
0.0

Brain (whole)
0.0
Liver (fetal)
0.0

Brain (amygdala)
0.0
Liver ca. (hepatoblast)
0.0

HepG2

Brain (cerebellum)
0.0
Lung
0.0

Brain (hippocampus)
0.0
Lung (fetal)
0.0

Brain (substantia nigra)
0.0
Lung ca. (small cell) LX-1
0.0

Brain (thalamus)
0.0
Lung ca. (small cell)
0.0

NCI-H69

Cerebral Cortex
0.0
Lung ca. (s.cell var.)
0.0

SHP-77

Spinal cord
0.0
Lung ca. (large cell)NCI-
0.0

H460

glio/astro U87-MG
0.0
Lung ca. (non-sm. cell)
0.0

A549

glio/astro U-118-MG
0.0
Lung ca. (non-s.cell)
0.0

NCI-H23

astrocytoma SW1783
0.0
Lung ca. (non-s.cell)
0.0

HOP-62

neuro*; met SK-N-AS
0.0
Lung ca. (non-s.cl) NCI-
0.0

H522

astrocytoma SF-539
0.1
Lung ca. (squam.) SW
0.0

900

astrocytoma SNB-75
100.0
Lung ca. (squam.) NCI-
0.0

H596

glioma SNB-19
0.0
Mammary gland
0.0

glioma U251
0.0
Breast ca.* (pl.ef) MCF-7
0.0

glioma SF-295
0.0
Breast ca.* (pl.ef) MDA-
0.0

MB-231

Heart (fetal)
0.0
Breast ca.* (pl.ef) T47D
0.0

Heart
0.0
Breast ca. BT-549
0.0

Skeletal muscle (fetal)
0.0
Breast ca. MDA-N
0.0

Skeletal muscle
0.0
Ovary
0.0

Bone marrow
0.0
Ovarian ca. OVCAR-3
0.0

Thymus
0.0
Ovarian ca. OVCAR-4
0.0

Spleen
0.0
Ovarian ca. OVCAR-5
0.0

Lymph node
0.0
Ovarian ca. OVCAR-8
0.0

Colorectal
0.0
Ovarian ca. IGROV-1
0.0

Stomach
0.0
Ovarian ca.* (ascites)
0.0

SK-OV-3

Small intestine
0.0
Uterus
0.0

Colon ca. SW480
0.0
Plancenta
0.0

Colon ca.* SW620(SW480
0.0
Prostate
0.0

met)

Colon ca. HT29
0.0
Prostate ca.* (bone
0.0

met)PC-3

Colon ca. HCT-116
0.0
Testis
0.0

Colon ca. CaCo-2
0.0
Melanoma Hs688(A).T
0.0

Colon ca.
0.0
Melanoma* (met)
0.0

tissue(ODO3866)

Hs688(B).T

Colon ca. HCC-2998
0.0
Melanoma UACC-62
0.0

Gastric ca.* (liver met)
0.0
Melanoma M14
0.0

NCI-N87

Bladder
0.1
Melanoma LOX IMVI
0.0

Trachea
0.0
Melanoma* (met) SK-
0.0

MEL-5

Kidney
0.0
Adipose
0.0

[0924] Panel 1.3D Summary: Ag2507 Significant expression of the CG50271-01 gene is seen exclusively in an astrocytoma cell line (CT=26.8). Therefore, expression of this gene may be used to distinguish astrocytoma cell lines from the other samples on this panel. Furthermore, therapeutic modulation of the activity of the GPCR encoded by this gene may be beneficial in the treatment of astrocytoma.

[0925] Panel 4D Summary: Ag1716/Ag2507 Expression of the CG50271-01 gene is low/undetectable (CTs>35) across all of the samples on this panel (data not shown).

[0926] BA. CG57497-01: Olfactory Receptor

[0927] Expression of gene CG57497-01 was assessed using the primer-probe set Ag5287, described in Table BAA. Results of the RTQ-PCR runs are shown in Table BAB.

242TABLE BAAProbe Name Ag5287StartSEQ IDPrimersSequencesLengthPositionNO:Forward5′-accctgccatcatgactgta-3′20399451ProbeTET-5′-tggtaagctggtgtctttctgttggc-3′-TAMRA26427452Reverse5′-aatgggaattgggtatccaa-3′20465453

[0928]

243

TABLE BAB

General_screening_panel_v1.5

Rel. Exp. (%) Ag5287, Run

Rel. Exp. (%) Ag5287, Run

Tissue Name
233239011
Tissue Name
233239011

Adipose
10.2
Renal ca. TK-10
6.8

Melanoma* Hs688(A).T
8.8
Bladder
4.4

Melanoma* Hs688(B).T
0.3
Gastric ca. (liver met.)
35.1

NCI-N87

Melanoma* M14
0.0
Gastric ca. KATO III
0.0

Melanoma* LOXIMVI
0.0
Colon ca. SW-948
0.0

Melanoma* SK-MEL-5
0.0
Colon ca. SW480
8.0

Squamous cell
2.4
Colon ca.* (SW480 met)
2.9

carcinoma SCC-4

SW620

Testis Pool
9.5
Colon ca. HT29
0.0

Prostate ca.* (bone met)
2.8
Colon ca. HCT-116
14.7

PC-3

Prostate Pool
2.4
Colon ca. CaCo-2
2.1

Placenta
0.0
Colon cancer tissue
0.0

Uterus Pool
3.4
Colon ca. SW1116
0.0

Ovarian ca. OVCAR-3
14.0
Colon ca. Colo-205
0.0

Ovarian ca. SK-OV-3
18.8
Colon ca. SW-48
0.0

Ovarian ca. OVCAR-4
0.0
Colon Pool
11.9

Ovarian ca. OVCAR-5
11.2
Small Intestine Pool
30.8

Ovarian ca. IGROV-1
5.3
Stomach Pool
37.6

Ovarian ca. OVCAR-8
5.9
Bone Marrow Pool
10.7

Ovary
9.4
Fetal Heart
3.3

Breast ca. MCF-7
5.7
Heart Pool
7.4

Breast ca. MDA-MB-
5.3
Lymph Node Pool
11.4

231

Breast ca. BT 549
7.0
Fetal Skeletal Muscle
3.5

Breast ca. T47D
2.7
Skeletal Muscle Pool
0.0

Breast ca. MDA-N
0.0
Spleen Pool
3.1

Breast Pool
16.7
Thymus Pool
18.6

Trachea
4.9
CNS cancer (glio/astro)
2.8

U87-MG

Lung
7.7
CNS cancer (glio/astro) U-
8.8

118-MG

Fetal Lung
5.9
CNS cancer (neuro;met)
7.1

SK-N-AS

Lung ca. NCI-N417
0.0
CNS cancer (astro) SF-539
0.0

Lung ca. LX-1
4.8
CNS cancer (astro) SNB-75
9.5

Lung ca. NCI-H146
2.0
CNS cancer (glio) SNB-19
0.0

Lung ca. SHP-77
0.0
CNS cancer (glio) SF-295
12.0

Lung ca. A549
10.7
Brain (Amygdala) Pool
0.0

Lung ca. NCI-H526
0.0
Brain (cerebellum)
0.0

Lung ca. NCI-H23
29.9
Brain (fetal)
5.9

Lung ca. NCI-H460
45.1
Brain (Hippocampus) Pool
7.9

Lung ca. HOP-62
3.4
Cerebral Cortex Pool
4.0

Lung ca. NCI-H522
9.0
Brain (Substantia nigra)
0.0

Pool

Liver
0.0
Brain (Thalamus) Pool
1.7

Fetal Liver
0.0
Brain (whole)
3.7

Liver ca. HepG2
0.0
Spinal Cord Pool
0.0

Kidney Pool
100.0
Adrenal Gland
0.0

Fetal Kidney
55.9
Pituitary gland Pool
3.0

Renal ca. 786-0
0.8
Salivary Gland
0.0

Renal ca. A498
0.0
Thyroid (female)
0.0

Renal ca. ACHN
3.2
Pancreatic ca. CAPAN2
15.5

Renal ca. UO-31
5.3
Pancreas Pool
15.4

[0929] CNS_neurodegeneration_v1.0 Summary: Ag5287 Expression of the CG57497-01 gene is low/undetectable in all samples in this panel (CTs>34.5). (Data not shown.)

[0930] General_screening_panel_v1.5 Summary: Ag5287 Highest expression of the CG57497-01 gene is seen in the kidney (CT=30.6). Significant expression is also seen in the fetal kidney. Thus, expression of this gene could be used to distinguish kidney derived tissue from other tissue.

[0931] Among tissues with metabolic function, this gene is expressed at low but significant levels in the pancreas and adipose. This suggests that this gene product may be involved in the pathogenesis and/or treatment of disease in these organs, including obesity and diabetes.

[0932] There is also significant expression in cell lines derived from gastric, lung, ovarian, colon and brain cancers. Thus, expression of this gene may be used to differentiate between samples derived from these cancers and other tissues on this panel. Furthermore, expression of this gene may be useful to detect the presence of these cancers.

[0933] Panel 4.1D Summary: Ag5287 Expression of the CG57497-01 gene is low/undetectable in all samples in this panel (CTs>34.5). (Data not shown.)

[0934] BB. GMAC024399_A—1: Olfactory Receptor

[0935] Expression of gene GMAC024399_A—1 was assessed using the primer-probe sets Ag1789 and Ag1714, described in Tables BBA and BBB. Results of the RTQ-PCR runs are shown in Table BBC. Please note that GMAC024399_A—1 was previously designated GMAC024399_A.

244TABLE BBAProbe Name Ag1789StartSEQ IDPrimersSequencesLengthPositionNO:Forward5′-gtgggtaacagcctcatagtca-3′22121454ProbeTET-5′-tggaccctcacctacactctcctatg-3′-TAMRA26155455Reverse5′-tgaaagattggtaagcaggaaa-3′22183456

[0936]

245

TABLE BBB

Probe Name Ag1714

Start
SEQ ID

Primers
Sequences
Length
Position
NO:

Forward
5′-gtgggtaacagcctcatagtca-3′
22
121
457

Probe
TET-5′-tggaccctcacctacactctcctatg-3′-TAMRA
26
155
458

Reverse
5′-tgaaagattggtaagcaggaaa-3′
22
183
459

[0937]

246

TABLE BBC

Panel 4D

Rel. Exp. (%)
Rel. Exp. (%)

Rel. Exp. (%)
Rel. Exp. (%)

Ag1714, Run
Ag1789, Run

Ag1714, Run
Ag1789, Run

Tissue Name
165330745
165809174
Tissue Name
165330745
165809174

Secondary Th1 act
0.0
0.0
HUVEC IL-1beta
0.0
0.0

Secondary Th2 act
0.0
0.0
HUVEC IFN gamma
0.0
0.0

Secondary Tr1 act
0.0
0.0
HUVEC TNF alpha +
0.0
0.0

IFN gamma

Secondary Th1 rest
0.0
0.0
HUVEC TNF alpha +
0.0
0.0

IL4

Secondary Th2 rest
0.0
0.0
HUVEC IL-11
0.0
0.0

Secondary Tr1 rest
17.3
0.0
Lung Microvascular
0.0
0.0

EC none

Primary Th1 act
0.0
0.0
Lung Microvascular
0.0
0.0

EC TNF alpha + IL-

1beta

Primary Th2 act
27.9
0.0
Microvascular
0.0
0.0

Dermal EC none

Primary Tr1 act
0.0
0.0
Microvascular Dermal
0.0
0.0

EC TNF alpha + IL-

1beta

Primary Th1 rest
0.0
0.0
Bronchial epithelium
0.0
0.0

TNF alpha + IL1beta

Primary Th2 rest
0.0
0.0
Small airway
0.0
0.0

epithelium none

Primary Tr1 rest
0.0
0.0
Small airway
0.0
0.0

epithelium TNF

alpha + IL-1beta

CD45RA CD4
0.0
0.0
Coronery artery SMC
0.0
0.0

lymphocyte act

rest

CD45RO CD4
0.0
0.0
Coronery artery SMC
0.0
0.0

lymphocyte act

TNF alpha + IL-1beta

CD8 lymphocyte act
0.0
0.0
Astrocytes rest
0.0
0.0

Secondary CD8
0.0
0.0
Astrocytes TNF
0.0
0.0

lymphocyte rest

alpha + IL-1beta

Secondary CD8
0.0
0.0
KU-812 (Basophil)
0.0
0.0

lymphocyte act

rest

CD4 lymphocyte
0.0
0.0
KU-812 (Basophil)
0.0
0.0

none

PMA/ionomycin

2ry
7.0
0.0
CCD1106
0.0
0.0

Th1/Th2/Tr1_anti-

(Keratinocytes) none

CD95 CH11

LAK cells rest
0.0
0.0
CCD1106
0.0
0.0

(Keratinocytes)

TNF alpha + IL-1beta

LAK cells IL-2
0.0
0.0
Liver cirrhosis
100.0
100.0

LAK cells IL-2 +
0.0
0.0
Lupus kidney
0.0
0.0

IL-12

LAK cells IL-2 +
0.0
0.0
NCI-H292 none
0.0
0.0

IFN gamma

LAK cells IL-2 +
0.0
0.0
NCI-H292 IL-4
0.0
0.0

IL-18

LAK cells
0.0
0.0
NCI-H292 IL-9
0.0
0.0

PMA/ionomycin

NK Cells IL-2 rest
0.0
0.0
NCI-H292 IL-13
0.0
0.0

Two Way MLR 3
0.0
0.0
NCI-H292 IFN
0.0
0.0

day

gamma

Two Way MLR 5
0.0
0.0
HPAEC none
0.0
0.0

day

Two Way MLR 7
0.0
0.0
HPAEC TNF alpha +
0.0
0.0

day

IL-1beta

PBMC rest
0.0
0.0
Lung fibroblast none
0.0
0.0

PBMC PWM
0.0
0.0
Lung fibroblast TNF
7.9
0.0

alpha + IL-1beta

PBMC PHA-L
0.0
0.0
Lung fibroblast IL-4
0.0
0.0

Ramos (B cell) none
0.0
0.0
Lung fibroblast IL-9
0.0
0.0

Ramos (B cell)
0.0
0.0
Lung fibroblast IL-13
0.0
0.0

ionomycin

B lmphocytes PWM
0.0
0.0
Lung fibroblast IFN
0.0
0.0

gamma

B lymphocytes
0.0
0.0
Dermal fibroblast
0.0
0.0

CD40L and IL-4

CCD1070 rest

EOL-1 dbcAMP
0.0
0.0
Dermal fibroblast
0.0
0.0

CCD1070 TNF alpha

EOL-1 dbcAMP
0.0
0.0
Dermal fibroblast
0.0
0.0

PMA/ionomycin

CCD1070 IL-1beta

Dendritic cells none
0.0
0.0
Dermal fibroblast IFN
0.0
0.0

gamma

Dendritic cells LPS
0.0
0.0
Dermal fibroblast IL-4
38.2
0.0

Dendritic cells anti-
0.0
0.0
IBD Colitis 2
60.3
51.8

CD40

Monocytes rest
0.0
0.0
IBD Crohn's
55.9
10.4

Monocytes LPS
0.0
2.1
Colon
0.0
3.7

Macrophages rest
22.5
0.0
Lung
0.0
0.0

Macrophages LPS
0.0
0.0
Thymus
0.0
1.4

HUVEC none
0.0
0.0
Kidney
0.0
1.6

HUVEC starved
0.0
0.0

[0938] Panel 1.3D Summary: Ag1789 Expression of the GMAC024399_A—1 gene is low/undetectable (CTs>35) across all of the samples on this panel (data not shown).

[0939] Panel 2.2 Summary: Ag1789 Expression of the GMAC024399_A—1 gene is low/undetectable (CTs>35) across all of the samples on this panel (data not shown).

[0940] Panel 4D Summary: Ag1714/Ag1789 Results from two experiments using an identical probe/primer set show reasonable agreement. Significant expression of the GMAC024399_A—1 gene is detected in a liver cirrhosis sample. Furthermore, expression of this gene is not detected in normal liver in Panel 1.3D, suggesting that its expression is unique to liver cirrhosis. This gene encodes a putative GPCR; therefore, antibodies or small molecule therapeutics could reduce or inhibit fibrosis that occurs in liver cirrhosis. In addition, antibodies to this putative GPCR could also be used for the diagnosis of liver cirrhosis (Mark et al., G protein modulation of recombinant P/Q-type calcium channels by regulators of G protein signalling proteins. J. Physiol. 528 Pt 1:65-77, 2000).

[0941] BC. SC122737711_A—1: Olfactory Receptor

[0942] Expression of gene SC122737711_A—1 was assessed using the primer-probe sets Ag1525, Ag295, Ag1628 and Ag2436, described in Tables BCA, BCB, BCC and BCD. Results of the RTQ-PCR runs are shown in Tables BCE, BCF and BCG. Please note that SC122737711_A—1 was previously designated SC122737711_A.

247TABLE BCAProbe Name Ag1525StartSEQ IDPrimersSequencesLengthPositionNO:Forward5′-gacatggcacctgttatcaagt-3′22541460ProbeTET-5′-cctgcactgacacccatgtgaaagag-3′-TAMRA26566461Reverse5′-ggatgctgaggctaaataaagc-3′22595462

[0943]

248

TABLE BCB

Probe Name Ag295

Start
SEQ ID

Primers
Sequences
Length
Position
NO:

Forward
5′-gtgccacccagctgttcttt-3′
20
293
463

Probe
TET-5′-ttggctttgcttgcaccaactgcc-3′-TAMRA
24
317
464

Reverse
5′-cgatcatatcccatcacagcaa-3′
22
347
465+TZ,1/44

[0944]

249

TABLE BCC

Probe Name Ag1628

Start
SEQ ID

Primers
Sequences
Length
Position
NO:

Forward
5′-gacatggcacctgttatcaagt-3′
22
541
466

Probe
TET-5′-cctgcactgacacccatgtgaaagag-3′-TAMRA
26
566
467

Reverse
5′-ggatgctgaggctaaataaagc-3′
22
595
468

[0945]

250

TABLE BCD

Probe Name Ag2436

Start
SEQ ID

Primers
Sequences
Length
Position
NO:

Forward
5′-ggtggcagtgacctacaca-3′
19
819
469

Probe
TET-5′-tcctcttgtctacagtctgaggaacaa-3′-TAMRA
27
858
470

Reverse
5′-ccaagaactcttttcaatgca-3′
21
897
471

[0946]

251

TABLE BCE

Panel 1.2

Rel.

Rel.

Exp. (%)
Rel.
Rel.

Exp. (%)
Rel.
Rel.

Ag1525,
Exp. (%)
Exp. (%)

Ag1525,
Exp. (%)
Exp. (%)

Run
Ag295, Run
Ag295, Run

Run
Ag295, Run
Ag295, Run

Tissue Name
142140914
119216697
139460066
Tissue Name
142140914
119216697
139460066

Endothelial
0.0
0.5
0.0
Renal ca.
0.0
3.2
0.0

cells

786-0

Heart (Fetal)
1.4
0.5
0.0
Renal ca.
0.5
0.6
0.0

A498

Pancreas
0.0
1.5
0.0
Renal ca.
0.0
0.8
0.0

RXF 393

Pancreatic ca.
0.0
0.5
0.0
Renal ca.
2.3
0.3
0.0

CAPAN 2

ACHN

Adrenal Gland
0.0
1.3
0.0
Renal ca. UO-
5.6
0.5
0.8

31

Thyroid
0.0
0.5
0.0
Renal ca. TK-
9.8
0.8
0.0

10

Salivary gland
0.6
0.2
0.0
Liver
0.0
0.4
0.0

Pituitary gland
0.0
0.2
0.0
Liver (fetal)
0.0
1.6
0.0

Brain (fetal)
0.0
1.8
0.0
Liver ca.
0.0
0.3
0.0

(hepatoblast)

HepG2

Brain (whole)
0.0
1.3
0.0
Lung
0.0
0.6
0.0

Brain
0.0
1.0
0.0
Lung (fetal)
0.0
19.6
0.0

(amygdala)

Brain
0.0
1.4
0.0
Lung ca.
0.0
1.0
0.0

(cerebellum)

(small cell)

LX-1

Brain
0.0
1.6
0.0
Lung ca.
50.7
2.8
32.1

(hippocampus)

(small cell)

NCI-H69

Brain
0.0
1.1
0.0
Lung ca.
0.5
2.6
0.0

(thalamus)

(s.cell var.)

SHP-77

Cerebral Cortex
0.0
2.0
0.0
Lung ca.
10.7
2.4
0.0

(large

cell)NCI-

H460

Spinal cord
0.0
0.4
0.0
Lung ca.
20.3
0.6
2.5

(non-sm. cell)

A549

glio/astro U87-
0.0
0.2
0.0
Lung ca.
0.0
2.6
0.0

MG

(non-s.cell)

NCI-H23

glio/astro U-
0.0
0.4
0.0
Lung ca.
3.7
0.6
1.4

118-MG

(non-s.cell)

HOP-62

astrocytoma
3.3
0.6
0.0
Lung ca.
2.4
0.6
0.0

SW1783

(non-s.cl)

NCI-H522

neuro*; met
1.9
0.2
0.0
Lung ca.
4.8
0.4
0.0

SK-N-AS

(squam.) SW

900

astrocytoma
0.0
0.8
0.0
Lung ca.
47.3
2.5
42.9

SF-539

(squam.)

NCI-H596

astrocytoma
0.0
0.3
0.0
Mammary
0.0
2.0
0.0

SNB-75

gland

glioma SNB-19
5.2
0.7
0.0
Breast ca.*
87.1
1.6
62.4

(pl.ef) MCF-7

glioma U251
4.1
0.9
0.0
Breast ca.*
0.0
0.4
0.0

(pl.ef) MDA-

MB-231

glioma SF-295
0.0
0.2
0.0
Breast ca.*
36.3
1.2
18.9

(pl. ef) T47D

Heart
0.0
1.2
0.0
Breast ca.
7.5
1.0
0.0

BT-549

Skeletal Muscle
0.0
0.2
0.0
Breast ca.
12.7
0.4
5.1

MDA-N

Bone marrow
0.0
2.4
0.0
Ovary
0.0
0.3
0.0

Thymus
100.0
100.0
94.6
Ovarian ca.
4.2
0.1
0.0

OVCAR-3

Spleen
0.0
0.1
0.0
Ovarian ca.
1.2
0.1
0.0

OVCAR-4

Lymph node
0.0
0.6
0.0
Ovarian ca.
55.1
2.8
100.0

OVCAR-5

Colorectal
3.3
0.9
14.0
Ovarian ca.
1.8
0.1
0.0

Tissue

OVCAR-8

Stomach
0.8
0.3
0.0
Ovarian ca.
7.6
0.1
2.7

IGROV-1

Small intestine
1.1
0.4
0.0
Ovarian ca.
13.1
0.2
0.0

(ascites) SK-

OV-3

Colon ca.
0.0
0.1
0.0
Uterus
0.0
0.2
0.0

SW480

Colon ca.*
0.0
0.2
0.0
Placenta
0.0
0.3
0.0

SW620

(SW480 met)

Colon ca. HT29
10.2
0.4
0.0
Prostate
0.0
0.4
0.0

Colon ca. HCT-
0.0
0.1
0.0
Prostate ca.*
0.4
0.8
0.0

116

(bone met)

PC-3

Colon ca.
1.1
0.3
0.0
Testis
0.0
29.5
0.0

CaCo-2

Colon ca.
25.5
4.2
15.5
Melanoma
0.0
1.4
0.0

Tissue

Hs688(A).T

(ODO3866)

Colon ca.
0.0
1.4
0.0
Melanoma*
2.9
1.0
0.0

HCC-2998

(met)

Hs688(B).T

Gastric ca.*
0.8
0.2
0.0
Melanoma
0.0
1.7
0.0

(liver met)

UACC-62

NCI-N87

Bladder
8.4
1.4
0.0
Melanoma
19.5
2.1
40.1

M14

Trachea
0.0
0.9
0.0
Melanoma
0.0
0.8
0.0

LOX IMVI

Kidney
0.0
0.2
0.0
Melanoma*
0.0
2.2
0.0

(met) SK-

MEL-5

Kidney (fetal)
0.0
1.8
0.0

[0947]

252

TABLE BCF

Panel 1.3D

Rel.
Rel.

Rel.
Rel.

Exp. (%)
Exp. (%)
Rel.

Exp. (%)
Exp. (%)
Rel.

Ag1628,
Ag2436,
Exp. (%)

Ag1628,
Ag2436,
Exp. (%)

Run
Run
Ag295, Run

Run
Run
Ag295, Run

Tissue Name
165924465
166108055
151966903
Tissue Name
165924465
166108055
151966903

Liver
0.0
0.0
0.0
Kidney (fetal)
0.0
0.0
0.0

adenocarcinoma

Pancreas
0.0
0.0
0.0
Renal ca.
15.6
0.0
0.0

786-0

Pancreatic ca.
0.0
0.0
2.9
Renal ca.
0.0
0.0
0.0

CAPAN 2

A498

Adrenal gland
0.0
0.0
3.4
Renal ca.
6.6
0.0
0.0

RXF 393

Thyroid
0.0
0.0
0.0
Renal ca.
0.0
0.0
0.0

ACHN

Salivary gland
0.0
0.0
0.0
Renal ca.
0.0
0.0
0.0

UO-31

Pituitary gland
0.0
0.0
0.0
Renal ca. TK-
0.0
0.0
0.0

10

Brain (fetal)
0.0
0.0
0.0
Liver
0.0
0.0
10.2

Brain (whole)
0.0
0.0
0.0
Liver (fetal)
0.0
0.0
0.0

Brain (amygdala)
0.0
0.0
0.0
Liver ca.
0.0
0.0
0.0

(hepatoblast)

HepG2

Brain
0.0
0.0
0.0
Lung
0.0
0.0
0.0

(cerebellum)

Brain
0.0
0.0
0.0
Lung (fetal)
0.0
0.0
0.0

(hippocampus)

Brain (substantia
0.0
0.0
0.0
Lung ca.
0.0
0.0
0.0

nigra)

(small cell)

LX-1

Brain (thalamus)
0.0
0.0
0.0
Lung ca.
0.0
0.0
0.0

(small cell)

NCI-H69

Cerebral Cortex
0.0
0.0
0.0
Lung ca.
0.0
0.0
0.0

(s.cell var.)

SHP-77

Spinal cord
0.0
0.0
0.0
Lung ca.
0.0
0.0
3.6

(large

cell)NCI-

H460

glio/astro U87-
0.0
0.0
0.0
Lung ca.
0.0
0.0
0.0

MG

(non-sm. cell)

A549

glio/astro U-118-
0.0
0.0
0.0
Lung ca.
0.0
0.0
0.0

MG

(non-s.cell)

NCI-H23

astrocytoma
0.0
0.0
0.0
Lung ca.
0.0
0.0
0.0

SW1783

(non-s.cell)

HOP-62

neuro*; met SK-
0.0
0.0
0.0
Lung ca.
0.0
0.0
0.0

N-AS

(non-s.cl)

NCI-H522

astrocytoma SF-
0.0
0.0
0.0
Lung ca.
0.0
4.6
0.0

539

(squam.) SW

900

astrocytoma SNB-
0.0
0.0
2.5
Lung ca.
5.7
0.0
0.0

75

(squam.)

NCI-H596

glioma SNB-19
0.0
0.0
2.5
Mammary
0.0
0.0
0.0

gland

glioma U251
0.0
0.0
0.0
Breast ca.*
15.5
2.5
10.6

(pl.ef) MCF-7

glioma SF-295
0.0
0.0
0.0
Breast ca.*
0.0
0.0
0.0

(pl.ef) MDA-

MB-231

Heart (fetal)
0.0
0.0
0.0
Breast ca.*
0.0
0.0
0.0

(pl.ef) T47D

Heart
0.0
0.0
0.0
Breast ca.
0.0
0.0
4.0

BT-549

Skeletal muscle
0.0
0.0
1.9
Breast ca.
0.0
0.0
0.0

(fetal)

MDA-N

Skeletal muscle
0.0
0.0
0.0
Ovary
0.0
0.0
0.0

Bone marrow
0.0
0.0
0.0
Ovarian ca.
0.0
0.0
0.0

OVCAR-3

Thymus
60.7
100.0
100.0
Ovarian ca.
0.0
0.0
0.0

OVCAR-4

Spleen
100.0
0.0
0.0
Ovarian ca.
0.0
0.0
0.0

OVCAR-5

Lymph node
0.0
0.0
0.0
Ovarian ca.
0.0
0.0
0.0

OVCAR-8

Colorectal
0.0
0.0
13.8
Ovarian ca.
0.0
0.0
2.7

IGROV-1

Stomach
0.0
0.0
5.2
Ovarian ca.*
15.4
0.0
3.2

(ascites) SK-

OV-3

Small intestine
0.0
0.0
0.0
Uterus
0.0
0.0
0.0

Colon ca. SW480
0.0
0.0
0.0
Plancenta
0.0
0.0
0.0

Colon ca.*
0.0
0.0
0.0
Prostate
0.0
0.0
0.0

SW620(SW480

met)

Colon ca. HT29
0.0
0.0
2.1
Prostate ca.*
0.0
0.0
0.0

(bone

met)PC-3

Colon ca. HCT-
0.0
0.0
0.0
Testis
0.0
0.0
3.4

116

Colon ca. CaCo-2
0.0
0.0
0.0
Melanoma
0.0
0.0
0.0

Hs688(A).T

Colon ca.
0.0
0.0
0.0
Melanoma*
0.0
0.0
0.0

tissue(ODO3866)

(met)

Hs688(B).T

Colon ca. HCC-
0.0
0.0
0.0
Melanoma
0.0
0.0
0.0

2998

UACC-62

Gastric ca.* (liver
0.0
0.0
0.0
Melanoma
0.0
0.0
0.0

met) NCI-N87

M14

Bladder
0.0
0.0
0.0
Melanoma
0.0
0.0
0.0

LOX IMVI

Trachea
0.0
0.0
0.0
Melanoma*
0.0
0.0
0.0

(met) SK-

MEL-5

Kidney
0.0
0.0
0.0
Adipose
0.0
0.0
0.0

[0948]

253

TABLE BCG

Panel 4D

Rel.
Rel.
Rel.

Rel.
Rel.
Rel.

Exp. (%)
Exp. (%)
Exp. (%)

Exp. (%)
Exp. (%)
Exp. (%)

Ag1628,
Ag2436,
Ag295,

Ag1628,
Ag2436,
Ag295,

Run
Run
Run

Run
Run
Run

Tissue Name
165762886
164325529
138033693
Tissue Name
165762886
164325529
138033693

Secondary Th1 act
0.0
0.0
0.0
HUVEC IL-
0.0
0.0
0.0

1beta

Secondary Th2 act
0.0
0.0
0.0
HUVEC IFN
0.0
0.0
0.0

gamma

Secondary Tr1 act
0.0
0.0
0.5
HUVEC TNF
0.0
0.0
0.0

alpha + IFN

gamma

Secondary Th1
0.0
0.0
0.0
HUVEC TNF
0.0
0.0
0.0

rest

alpha + IL4

Secondary Th2
0.0
0.0
0.0
HUVEC IL-11
0.0
0.0
0.0

rest

Secondary Tr1 rest
0.0
0.0
0.0
Lung
0.0
0.0
0.0

Microvascular

EC none

Primary Th1 act
0.0
0.0
0.0
Lung
0.0
0.0
0.0

Microvascular

EC TNF alpha +

IL-1beta

Primary Th2 act
0.0
0.0
1.3
Microvascular
0.0
0.0
0.0

Dermal EC none

Primary Tr1 act
0.0
0.0
0.0
Microsvasular
0.0
0.0
0.0

Dermal EC

TNF alpha + IL-

1beta

Primary Th1 rest
0.0
0.0
1.4
Bronchial
0.0
0.0
0.0

epithelium

TNF alpha +

IL1beta

Primary Th2 rest
1.2
0.0
0.0
Small airway
0.0
0.0
0.0

epithelium none

Primary Tr1 rest
0.0
0.0
0.0
Small airway
0.0
0.0
1.3

epithelium

TNF alpha + IL-

1beta

CD45RA CD4
0.0
0.0
0.0
Coronery artery
0.0
0.0
0.0

lymphocyte act

SMC rest

CD45RO CD4
0.0
0.0
0.0
Coronery artery
0.0
0.0
0.0

lymphocyte act

SMC TNF

alpha + IL-1beta

CD8 lymphocyte
0.0
0.0
0.0
Astrocytes rest
0.0
0.0
0.0

act

Secondary CD8
0.0
0.0
0.0
Astrocytes
0.0
0.0
0.0

lymphocyte rest

TNF alpha + IL-

1beta

Secondary CD8
0.0
0.0
0.0
KU-812
0.0
0.0
0.0

lymphocyte act

(Basophil) rest

CD4 lymphocyte
0.0
0.0
0.0
KU-812
0.0
0.0
0.0

none

(Basophil)

PMA/ionomycin

2ry
0.0
0.0
4.3
CCD1106
0.0
0.0
0.0

Th1/Th2/Tr1_anti-

(Keratinocytes)

CD95 CH11

none

LAK cells rest
0.0
0.0
0.0
CCD1106
0.0
0.0
0.0

(Keratinocytes)

TNF alpha + IL-

1beta

LAK cells IL-2
0.0
0.0
0.0
Liver cirrhosis
34.2
1.2
6.4

Lak cells IL-
0.0
0.0
0.0
Lupus kidney
0.0
0.0
0.0

2 + IL-12

LAK cells IL-
0.0
0.0
0.0
NCI-H292 none
0.0
0.0
0.0

2 + IFN gamma

LAK cells IL-2 +
0.0
0.0
0.0
NCI-H292 IL-4
0.0
0.0
0.0

IL-18

LAK cells
0.0
0.0
1.9
NCI-H292 IL-9
0.0
0.0
0.0

PMA/ionomycin

NK Cells IL-2 rest
0.0
0.0
0.0
NCI-H292 IL-13
0.0
0.7
0.0

Two Way MLR 3
0.0
0.0
0.0
NCI-H292 IFN
0.0
0.0
0.0

day

gamma

Two Way MLR 5
0.0
0.0
0.0
HPAEC none
0.0
0.0
0.0

day

Two Way MLR 7
0.0
0.0
0.0
HPAEC TNF
0.0
0.0
0.0

day

alpha + IL-1beta

PBMC rest
0.0
0.0
0.0
Lung fibroblast
0.0
0.0
0.0

none

PBMC PWM
0.0
0.0
1.3
Lung fibroblast
0.0
0.0
0.0

TNF alpha + IL-

1beta

PBMC PHA-L
0.0
0.0
0.0
Lung fibroblast
0.0
0.0
0.0

IL-4

Ramos (B cell)
0.0
0.0
0.0
Lung fibroblast
0.0
0.0
0.0

none

IL-9

Ramos (B cell)
0.0
0.0
0.0
Lung fibroblast
0.0
0.0
0.0

ionomycin

IL-13

B lymphocytes
0.0
0.0
0.0
Lung fibroblast
0.0
0.0
0.0

PWM

IFN gamma

B lymphocytes
0.0
0.0
0.0
Dermal
0.0
0.0
0.0

CD40L and IL-4

fibroblast

CCD1070 rest

EOL-1 dbcAMP
1.4
0.0
0.0
Dermal
0.0
0.0
0.0

fibroblast

CCD1070 TNF

alpha

EOL-1 dbcAMP
0.8
0.0
0.7
Dermal
0.0
0.0
0.0

PMA/ionomycin

fibroblast

CCD1070 IL-1

beta

Dendritic cells
0.0
0.0
0.0
Dermal
0.0
0.0
0.0

none

fibroblast IFN

gamma

Dendritic cells
0.0
0.0
0.0
Dermal
0.0
0.0
0.0

LPS

fibroblast IL-4

Dendritic cells
0.0
0.0
0.0
IBD Colitis 2
2.4
0.0
2.0

anti-CD40

Monocytes rest
0.0
0.0
0.0
IBD Crohn's
0.0
0.0
0.0

Monocytes LPS
0.0
0.0
0.0
Colon
0.6
0.0
0.0

Macrophages rest
0.0
0.0
0.0
Lung
0.0
0.0
0.0

Macrophages LPS
0.0
0.0
0.0
Thymus
0.0
0.0
0.0

HUVEC none
0.0
0.0
0.0
Kidney
100.0
100.0
100.0

HUVEC starved
0.0
0.0
0.0

[0949] Panel 1.2 Summary: Ag295 Expression of the SC122737711_A—1 gene in Panel 1.2 appears to be generally associated with normal tissues, with highest expression in thymus (CTs=29-32). Thus, the expression of this gene could be utilized to distinguish thymic tissue from other tissues. The gene is also moderately expressed in testis.

[0950] There is also high expression in cell lines derived from breast, ovarian and lung cancer cell lines. Thus, expression of this gene could be used to differentiate between these samples and other samples on this panel. Furthermore, expression of this gene could potentially be used as marker to detect the presence of these cancers.

[0951] Panel 1.3D Summary: Ag295/Ag1628/Ag2436 The SC122737711_A—1 gene is expressed consistently in the thymus in all three experiments using Panel 1.3D as well as in Panel 1.2. The SC 122737711_A—1 gene or the protein encoded for by this gene may be used as markers for thymic tissue. Antibodies raised against the protein encoded for by the SC122737711_A—1 gene could also be used as a tool to identify thymic tissue.

[0952] Panel 2D Summary: Ag295 Expression of the SC122737711_A—1 gene is low to undetectable in all samples on these panels (CT values>35). (Data not shown.)

[0953] Panel 4D Summary: Ag295/Ag1628/Ag2436 Three experiments show highest expression of the SC122737711_A—1 transcript is expressed in the kidney (CTs=30-32). The protein encoded for by the SC122737711_A—1 gene may therefore be involved in the development and homeostasis of the kidney. Thus, expression of this gene could be used as a marker of kidney tissue. Furthermore, therapeutic modulation of the expression or function of the protein encoded by this gene may be of use in maintaining and restoring function to the kidney during inflammation.

[0954] BD. CG50149-02: Olfactory Receptor

[0955] Expression of gene CG50149-02 was assessed using the primer-probe sets Ag2364 and Ag1725, described in Tables BDA and BDB. Results of the RTQ-PCR runs are shown in Table BDC.

254TABLE BDAProbe Name Ag2364StartSEQ IDPrimersSequencesLengthPositionNO:Forward5′-caacaagccacagagatagttg-3′22224472ProbeTET-5′-cttcttgcaggaagccctccagcatt-3′-TAMRA26198473Reverse5′-tctctacacctccgcagtgat-3′21171474

[0956]

255

TABLE BDB

Probe Name Ag1725

Start
SEQ ID

Primers
Sequences
Length
Position
NO:

Forward
5′-gctcaggtgacaactctcattc-3′
22
538
475

Probe
TET-5′-tgtgttctgcctcactattccttttgga-3′-TAMRA
28
564
476

Reverse
5′-caccacaattctggcataagat-3′
22
603
477

[0957]

256

TABLE BDC

Panel 4D

Rel. Exp. (%)
Rel. Exp. (%)

Rel. Exp. (%)
Rel. Exp. (%)

Ag1725, Run
Ag2364, Run

Ag1725, Run
Ag2364, Run

Tissue Name
165767161
162361133
Tissue Name
165767161
162361133

Secondary Th1 act
0.0
0.0
HUVEC IL-1beta
0.0
0.0

Secondary Th2 act
0.0
0.0
HUVEC IFN gamma
0.0
0.0

Secondary Tr1 act
0.0
0.0
HUVEC TNF alpha +
0.0
0.0

IFN gamma

Secondary Th1 rest
0.0
0.0
HUVEC TNF alpha +
0.0
0.0

IL4

Secondary Th2 rest
0.0
0.0
HUVEC IL-11
0.0
0.0

Secondary Tr1 rest
2.1
0.0
Lung Microvascular
2.3
0.0

EC none

Primary Th1 act
0.0
0.0
Lung Microvascular
0.0
0.0

EC TNFalpha + IL-

1beta

Primary Th2 act
0.0
0.0
Microvascular
0.0
0.0

Dermal EC none

Primary Tr1 act
0.0
0.0
Microsvasular Dermal
0.0
0.0

EC TNFalpha + IL-

1beta

Primary Th1 rest
0.0
0.0
Bronchial epithelium
0.0
0.0

TNFalpha + IL1beta

Primary Th2 rest
0.0
0.0
Small airway
0.0
0.0

epithelium none

Primary Tr1 rest
0.0
0.0
Small airway
0.0
0.0

epithelium TNFalpha +

IL-1beta

CD45RA CD4
0.0
0.0
Coronery artery SMC
0.0
0.0

lymphocyte act

rest

CD45RO CD4
0.0
0.0
Coronery artery SMC
0.0
0.0

lymphocyte act

TNFalpha + IL-1beta

CD8 lymphocyte act
0.0
0.0
Astrocytes rest
0.0
0.0

Secondary CD8
0.0
0.0
Astrocytes TNFalpha +
0.0
0.0

lymphocyte rest

IL-1beta

Secondary CD8
0.0
0.0
KU-812 (Basophil)
0.0
13.0

lymphocyte act

rest

CD4 lymphocyte
0.0
0.0
KU-812 (Basophil)
3.7
0.0

none

PMA/ionomycin

2ry
0.0
0.0
CCD1106
0.0
0.0

Th1/Th2/Tr1_anti-

(Keratinocytes) none

CD95 CH11

LAK cells rest
0.0
0.0
CCD1106
3.0
0.0

(Keratinocytes)

TNFalpha + IL-1beta

LAK cells IL-2
0.0
0.0
Liver cirrhosis
100.0
100.0

LAK cells IL-2 + IL-
0.0
0.0
Lupus kidney
2.0
0.0

12

LAK cells IL-2 + IFN
0.0
0.0
NCI-H292 none
0.0
0.0

gamma

LAK cells IL-2 + IL-
0.0
0.0
NCI-H292 IL-4
0.0
0.0

18

LAK cells
0.0
0.0
NCI-H292 IL-9
0.0
0.0

PMA/ionomycin

NK Cells IL-2 rest
0.0
0.0
NCI-H292 IL-13
0.0
0.0

Two Way MLR 3
0.0
0.0
NCI-H292 IFN
0.0
0.0

day

gamma

Two Way MLR 5
0.0
0.0
HPAEC none
0.0
0.0

day

Two Way MLR 7
0.0
0.0
HPAEC TNF alpha +
0.0
0.0

day

IL-1 beta

PBMC rest
0.0
0.0
Lung fibroblast none
0.0
0.0

PBMC PWM
0.0
0.0
Lung fibroblast TNF
0.0
0.0

alpha + IL-1 beta

PBMC PHA-L
0.0
0.0
Lung fibroblast IL-4
0.0
0.0

Ramos (B cell) none
0.0
0.0
Lung fibroblast IL-9
0.0
0.0

Ramos (B cell)
0.0
0.0
Lung fibroblast IL-13
0.0
0.0

ionomycin

B lymphocytes PWM
0.0
0.0
Lung fibroblast IFN
0.0
0.0

gamma

B lymphocytes
0.0
0.0
Dermal fibroblast
0.0
0.0

CD40L and IL-4

CCD1070 rest

EOL-1 dbcAMP
0.0
0.0
Dermal fibroblast
0.0
0.0

CCD1070 TNF alpha

EOL-1 dbcAMP
0.0
0.0
Dermal fibroblast
0.0
0.0

PMA/ionomycin

CCD1070 IL-1 beta

Dendritic cells none
0.0
0.0
Dermal fibroblast IFN
0.0
0.0

gamma

Dendritic cells LPS
0.0
0.0
Dermal fibroblast IL-4
0.0
12.0

Dendritic cells anti-
0.0
0.0
IBD Colitis 2
6.3
20.7

CD40

Monocytes rest
0.0
0.0
IBD Crohn's
2.9
0.0

Monocytes LPS
0.0
0.0
Colon
9.7
0.0

Macrophages rest
0.0
0.0
Lung
0.0
0.0

Macrophages LPS
0.0
0.0
Thymus
0.0
25.3

HUVEC none
0.0
0.0
Kidney
0.0
0.0

HUVEC starved
4.5
0.0

[0958] CNS_neurodegeneration_v1.0 Summary: Ag2364 Expression of the CG50149-02 gene is low/undetected in all samples in this panel. (Data not shown.)

[0959] Panel 1.3D Summary: Ag2364 Expression of the CG50149-02 gene is low/undetected in all samples in this panel. (Data not shown.)

[0960] Panel 2.2 Summary: Ag2364 Expression of the CG50149-02 gene is low/undetected in all samples in this panel. (Data not shown.)

[0961] Panel 4D Summary: Ag1725/Ag2364 The CG50149-02 transcript is only detected in liver cirrhosis. Furthermore, this transcript is not detected in normal liver in Panel 1.3D, suggesting that this gene expression is unique to liver cirrhosis. This gene encodes a putative GPCR; therefore, antibodies or small molecule therapeutics could reduce or inhibit fibrosis that occurs in liver cirrhosis. In addition, antibodies to this putative GPCR could also be used for the diagnosis of liver cirrhosis.

Equivalents

[0962] Although particular embodiments have been disclosed herein in detail, this has been done by way of example for purposes of illustration only, and is not intended to be limiting with respect to the scope of the appended claims, which follow. In particular, it is contemplated by the inventors that various substitutions, alterations, and modifications may be made to the invention without departing from the spirit and scope of the invention as defined by the claims. The choice of nucleic acid starting material, clone of interest, or library type is believed to be a matter of routine for a person of ordinary skill in the art with knowledge of the embodiments described herein. Other aspects, advantages, and modifications considered to be within the scope of the following claims.

Number	Date	Country
60256635	Dec 2000	US
60259743	Jan 2001	US
60299327	Jun 2001	US
60261498	Jan 2001	US
60263689	Jan 2001	US
60267464	Feb 2001	US
60271021	Feb 2001	US
60275946	Mar 2001	US
60278150	Mar 2001	US
60285718	Apr 2001	US
60312902	Aug 2001	US
60257876	Dec 2000	US
60260718	Jan 2001	US
60284591	Apr 2001	US

Novel proteins and nucleic acids encoding same

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