COMPOSITIONS AND METHODS FOR AUGMENTING ACTIVITY OF ONCOLYTIC VIRUSES

Abstract

Disclosed are compositions and methods for augmenting activity of oncolytic viruses. Virus activity is augmented by sensitizing cancer or tumour cells through modulation of the Endoplasmic Reticulum (ER) stress response pathway, for instance by introducing into a tumour cell an agent effective to modulate ER stress response and sensitize the tumour cell. The tumour cells are then contacted with an oncolytic virus in an amount effective to reduce viability of the sensitized tumour cell. The oncolytic virus is thereby rendered more effective at lysing or killing the sensitized tumour or cancer cells.

Description

FIELD OF INVENTION

The present invention relates to compositions and methods for augmenting activity of oncolytic viruses. In particular, oncolytic virus activity is augmented by sensitizing cancer or tumour cells through modulation of the Endoplasmic Reticulum (ER) stress response pathway.

BACKGROUND OF THE INVENTION

Despite major advances in the understanding of cancer over the last 50 years, it remains one of the most important health challenges worldwide. Innovative approaches are needed to complement current drug based therapeutic strategies, and oncolytic viruses represent one such promising tool in the fight against cancer.

Oncolytic viruses preferentially infect and lyse cancer cells. They have been shown to act: (i) by directly destroying tumour cells via their inherent cytolytic activity, and (ii) through modification to function as vectors for delivering genes expressing anticancer proteins to a tumour site.

One example of an oncolytic virus having cytolytic activity is ONYX-015. ONYX-015 is the commercial name of an adenovirus mutant (dll 520) that is replication-restricted in normal cells having a wild-type p53 gene. ONYX-015 has been shown to replicate and kill tumour cells lacking a functional p53.

When used as a vector, therapeutic or cytotoxic genes can be delivered by the oncolytic virus to a tumour site where the products of these genes either directly or indirectly inhibit tumour growth. A number of different genes have been used for such applications, including human cytokine genes, tumour suppressor genes, bacterial or viral prodrug-activating enzyme encoding genes (suicide genes) and genes which make the tumour mass more susceptible to radiation and chemotherapy.

A variety of different virus strains have been studied, including naturally occurring or genetically modified versions of adenovirus, herpes simplex virus (“HSV”), reovirus, poxvirus (e.g. vaccinia virus and Myxoma virus), vesicular stomatitis virus (“VSV”), poliovirus, Newcastle disease virus (“NDV”), and measles. However, such viruses often lack sufficient potency as monotherapies to be completely clinically effective anticancer agents.

In an effort to improve clinical efficacy, candidate viruses have been genetically engineered to express therapeutic transgenes, and have been combined with other common oncolytic therapies. While such studies are ongoing with encouraging success, there continues to be a need for ways to enhance potency and efficacy, and generally make oncolytic viruses more effective cancer therapeutics.

SUMMARY OF THE INVENTION

It is an object of the invention to provide improved compositions and methods for augmenting activity of oncolytic viruses.

The invention relates to a method of reducing viability of a tumor cell in a subject, comprising the steps of: (i) introducing into a tumor cell in the subject an agent effective to modulate endoplasmic reticulum (ER) stress response and sensitize the tumour cell to cytolytic activity of an oncolytic virus in the subject; and (ii) contacting the tumor cell with an oncolytic virus in an amount effective to reduce viability of the sensitized tumour cell, wherein viability of the tumor cell is reduced by the oncolytic virus. In a preferred embodiment, the oncolytic virus lyses or kills the sensitized tumour cell.

In a further embodiment, the invention relates to a method of modulating sensitivity of cancer cells to infection by an oncolytic virus, comprising introducing into a cancer cell an agent effective to modulate endoplasmic reticulum (ER) stress response and sensitize the cancer cell to cytolytic activity of the oncolytic virus, wherein the cancer cells are sensitized to infection by the oncolytic virus.

According to the methods described herein, the agent may be effective to enhance, diminish or inhibit the ER stress response in said subject. In one preferred embodiment, the agent is effective to inhibit the ER stress response in said subject. In a further embodiment, the agent may be a siRNA specific to ERN, ATF6, Derlin1, Derlin2 or SEC61. Alternately, the agent may be a molecule effective to modulate ERN, ATF6, Derlin1, Derlin2 or SEC61 signaling, for instance a molecule effective to block or enhance ERN, ATF6, Derlin1, Derlin2 or SEC61 signaling. It is also contemplated that the agent may be a modified oncolytic virus wherein the modification renders the oncolytic virus effective to modulate ER stress response and sensitize the tumour cell to cytolytic activity.

It is further envisioned that the tumour cells or cancer cells described herein may be any tumour or cancer cell that is susceptible to oncolytic virus infection and modulation of the ER stress response. Without wishing to be limiting in any way, such cancer and tumour cells may be colon cancer cells, lung cancer cells, liver cancer cells, prostate cancer cells, bladder cancer cells, neck and mouth cancer cells, breast cancer cells, glioblastoma cells, lymphoma cells, carcinoma cells, renal cell cancer cells, pancreatic cancer cells, ovarian cancer cells and any other such cancer or tumour cells known in the art.

In further embodiments, the oncolytic virus may be any oncolytic virus, such as but without wishing to be limiting, a native or modified herpes virus, Adenovirus, Adeno-associated virus, influenza virus, reovirus, rhabdovirus, Newcastle virus, vaccinia virus, poliovirus, measles virus, mumps virus, sindbis virus (SIN) or sendai virus (SV). In preferred embodiments the oncolytic virus may be a native or modified rhabdovirus, for example a native or modified vesicular stomatitis virus (VSV) or Maraba virus. By ‘modified’, it is meant that the virus is a mutant virus modified with a function-improving mutation to make the virus a more effective cancer or tumour cell lysing agent.

The invention further relates to a method of identifying a tumour cell sensitizing agent effective for sensitizing tumour cells to infection by an oncolytic virus. The method comprises: (i) providing a test molecule with putative endoplasmic reticulum (ER) stress response modulating activity, (ii) adding the test molecule to a sample of said tumor cells, (iii) contacting the tumor cells with the oncolytic virus, and (iv) comparing cytolytic activity of the oncolytic virus in the sample of tumour cells with the test molecule to activity of the oncolytic virus in a sample of tumour cells without the test molecule, wherein increased cytolytic activity of the oncolytic virus in the sample of tumour cells with the test molecule indicates the presence of a tumour cell sensitizing agent.

The test molecule described above may be any molecule suspected of having ER stress response modulating activity. Such molecules may be a small molecule, a protein, a nucleic acid, an antibody, or any other non-limiting example of a putative test molecule.

The invention further provides compounds effective to modulate endoplasmic reticulum (ER) stress response and sensitize a tumour cell to cytolytic activity of an oncolytic virus in a subject. Such compounds may be effective to inhibit the ER stress response in the subject. For instance, the compound may be a siRNA specific to ERN, ATF6, Derlin1, Derlin2 or SEC61, or a molecule effective to modulate ERN, ATF6, Derlin1, Derlin2 or SEC61 signaling. In another embodiment, the compounds may augment the ER stress response in the tumour to improve oncolytic therapy.

In an embodiment, the compound is effective to block or enhance ERN, ATF6, Derlin1, Derlin2 or SEC61 signaling. In a further embodiment, the compound is a protein, a small molecule, a nucleic acid, or an antibody.

The above-described compounds may also be formulated into a composition, for instance a pharmaceutical composition including a pharmaceutically acceptable carrier or excipient.

Also contemplated by the present invention is a modified oncolytic virus, wherein the modification renders the oncolytic virus effective to modulate endoplasmic reticulum (ER) stress response and sensitize a tumour cell to cytolytic activity of the oncolytic virus in a subject. The modified oncolytic virus may be effective to inhibit the ER stress response in the subject. In another embodiment, the modified oncolytic virus may augment the ER stress response in the tumour to improve oncolytic therapy.

In an embodiment, the oncolytic virus may be modified to include a nucleotide specific to ERN, ATF6, Derlin1, Derlin2 or SEC61 or a nucleotide encoding a molecule effective to modulate ERN, ATF6, Derlin1, Derlin2 or SEC61 signaling. In a further embodiment, the modified oncolytic virus may be effective to block or enhance ERN, ATF6, Derlin1, Derlin2 or SEC61 signaling. In a further embodiment, the molecule may be a protein or an antibody.

The above-described modified oncolytic virus may also be formulated into a composition, for instance a pharmaceutical composition including a pharmaceutically acceptable carrier or excipient.

The invention also relates to a method of sensitizing a tumor to cytolytic activity of an oncolytic virus, said method comprising inducing in a subject a mild stress to the endoplasmic reticulum (ER).

In a non-limiting embodiment, inducing the mild stress may comprise genetically disrupting an ER stress response gene, for instance a ER stress response gene such as IRE1/ERN, DERLIN, and ATF6. In another non-limiting embodiment, inducing the mild stress may comprise chemically inhibiting IRE1/ERN1. For instance, compound 2 (described herein) is administered to the subject to chemically inhibit IRE1/ERN1. In a further non-limiting embodiment, inducing the mild stress may comprise chemically inhibiting cyclophilins which blocks the function of chaperones in the ER. Without wishing to be limiting, Cyclosporin A can be administered to chemically inhibiting the cyclophilins. In another non-limiting embodiment, inducing the mild stress may comprise chemically inhibiting protein glycosylation and producing more unfolded proteins in the ER. For instance, but without wishing to be limiting, Tunicamycin can be administered to chemically inhibit protein glycosylation.

It is also to be understood that the above-described compound can be effective to induce an ER stress and render tumour cells susceptible to a virus infection. In addition, yet without wishing to be limiting, the compound can be effective to initiate caspase 2 mediated cell death in response to a virus infection, and render tumour cells susceptible to a virus infection.

In certain non-limiting embodiments, the compound may be one of the following

BRIEF DESCRIPTION OF THE DRAWINGS

These and other features of the invention will become more apparent from the following description in which reference is made to the appended drawings wherein:

FIG. 1 is a schematic describing the functional genetic screen used to identify host genes that modulate oncolytic virus therapy. Briefly, cells in 384 well format were reverse transfected with 10 nM siRNA pools from an arrayed genome wide library of siRNA (Dharmacon Inc. USA). After an incubation of 72 hours, cells were infected with Maraba virus at an MOI of 0.05 and further incubated for 48 hours. Cell viability was measured using Alamar Blue™ vital dye assay. Alternatively cells were washed, fixed and stained with hoechst nuclear stain and scored for cell number and nuclear morphology to assay cell viability;

FIG. 2 shows a summary of the genome-wide RNAi screen in two cancer cell lines: (A) Venn diagram indicating the number of genes scored as “hits” in the screen for each cell line. Hits are designated as genes with viability scores greater than two standard deviations of the negative control from the mean of the overall screen. In addition, genes with viability scores of one standard deviation of the negative control from the mean of RNAi alone screen were filtered from the list to remove false positives due to the effects of RNAi alone. (B) Bioinformatic analysis of the 485 hits common to both cell lines;

FIG. 3 illustrates results showing IRE1/ERN knockdown sensitizing of cancer cells to Maraba virus killing: (A) Cell lines were transfected with siRNA targeting IRE1alpha (ERN1) or IRE1beta (ERN2) or control in 96 well format. After 72 hours, wells were infected with Maraba virus at various MOIs. Viability was monitored by Alamar Blue™ viability assay following 48 h of infection. Experiments were performed in triplicate and plotted as the mean, with error bars representing the standard error. EC50 values were calculated for each condition and compared to the control to determine fold sensitization. (B) Summary of the results from (A) indicating the effects of loss of ERN mRNA on Maraba virus killing of tumour lines. (C) Western blot confirming the inhibition of ERN-1 (IRE1alpha) protein expression; and

FIG. 4 illustrates results showing Maraba virus activates the Unfolded Protein Response: (A) U373 and OVCAR-8 cells were treated with tunicamycin (5 μg/mL) or wild-type Maraba virus (MOI 5) for the indicated amounts of time. Total protein lysates were collected, and immunoblots performed using the indicated antibodies. F-ATF6, Full-length activated transcription factor 6. DG-ATF6, de-glycosylated ATF6. C-ATF-6, cleaved ATF6. P-EIF2α, phosphorylated eukaryotic initiation factor 2α. BIP/GRP78, immunoglobulin heavy chain binding protein/glucose regulated protein 78. XBP(s), spliced X-box binding protein 1. GAPDH, glyceraldehyde dehydrogenase. G, N, P and M indicate Maraba virus proteins. (B) U373 and OVCAR-8 cells were treated with tunicamycin (TM) and Maraba virus, as in (A). Total RNA was extracted and RT-PCR performed using XBP-specific primers. XBP(u), un-spliced XBP. XBP(s), spliced XBP.

FIG. 5 illustrates the results of the genome wide screen of Example 2, and the identification of ER stress response blockade as a potent sensitizer to rhabdovirus-mediated oncolysis. (A) Schematic representation of the screen. (B) Venn diagram outlining the number of overlapping hits, and a table (+=synthetic lethal, −=no interaction) and schematic diagram (hits outlined in red) illustrating key hits within the UPR and ERAD pathways. (C) Phase contrast images of U373 cells treated first with siRNA (72 h) followed by Maraba virus infection (MOI 5; 24 h). (D) Upper panels. Cell viability assays were performed 48 h after Maraba virus infection, which followed 72 h siRNA treatment. Western blots demonstrating protein knockdown are depicted (* denotes non-specific band). Lower panels. Cell viability assays were conducted 48 h after virus infection, in U373 cells ectopically expressing mouse ATF6α (or GFP control)±siRNA targeting human ATF6α (or NT control). (E) Representative tumour and normal cell lines were treated with siRNA targeting IRE1α for 72 h followed by Maraba virus (MOI 0.1). Cell viability assays were performed 48 h later. (F) Cell viability assays were performed on a panel of cancer-derived cell lines 48 h after Maraba virus infection (MOI 5), which followed UPR knockdown (72 h). Data is represented as “log sensitization”, which is defined as the reduction in the amount of Maraba virus (on a log₁₀ scale) required to obtain an EC₅₀. The degree of functionality of the interferon system is also plotted, with (−) indicating completely defective and (+++) indicating completely functional (N/A indicates cell lines not analyzed). (G) Cell viability was measured after 48 h infection with wild type or “double mutant” Maraba virus of wild type VSV (MOI 0.1), which followed 72 h siRNA treatment (*=p<0.05; #=p<0.01).

FIG. 6 illustrates that UPR knockdown sensitizes U373 cells towards Maraba virus mediated killing. U373 cells were treated with non- or IRE1α-targeting RNAi duplexes for 72 h prior to Maraba-WT infection. After 48 h, cell viability was assessed using Resazurin sodium salt (*=p<0.05).

FIG. 7 illustrates that acute inhibition of the ER stress response is not sufficient to sensitize tumour cells to viral oncolysis. (A) U373 cells were treated with the ER stressor tunicamycin (5 μg/mL) or Maraba virus (MOI 5). Total RNA was collected and RT-PCR for XBP1 splicing performed. (B) Cells were treated as in (A). Total cell lysates were collected and immunoblot analyses conducted (DG-ATG6α=deglycosylated ATF6α, due to the inhibitory effect of TM on glycosylation). (C) U373 cells were treated with putative IRE1α small molecule inhibitors (2 h) prior to tunicamycin treatment (4 h). Total RNA was collected and RT-PCR performed. RNAi targeting IRE1α (72 h) was used as a control. (D) U373 cells were treated with Compound 2 (50 uM) or controls for 4 or 48 h prior to Maraba virus infection. Cell viability assays were performed 48 h later. (E) Cells were treated as in (D) and combination index analyses performed. (F) U373 cells were treated with DMSO or cyclosporine (25 μM) for 4 or 48 h before Maraba virus infection. Cell viability was assessed 48 h later (*=p<0.05).

FIG. 8 shows ER preload rewires cancer cells for caspase 2-mediated apoptosis in response to oncolytic virus infection. (A) Cells were treated with siRNA for 0-72 h. Total cell lysates were collected followed by immunoblot analyses. (B) Cells were treated with siRNA (72 h), followed by infection with Maraba virus (MOI 5). Total cell lysates were collected and immunoblots performed. (C) Cells were treated as in (B) or treated with tunicamycin (5 ug/mL) for 24 h followed by 24 h “washout”, after which cells were infected with Maraba virus (MOI 5). Total cell lysates were collected at the indicated timepoints and immunoblots performed. (D-E) Cells were treated as in (B), and Western blots performed. (F) Schematic diagram depicting ER Preload model. Rhabdovirus infection of naïve tumour cells (Standard OVT) causes ER stress but fails to trigger a caspase 2 mediated apoptotic response (grayed out dormant response). Inhibiting UPR or ERAD (Combination ER/OV Therapy) induces an ER stress resulting in an adaptive response (rewiring) that predisposes tumour cells to undergo an alternative death program (caspase 2 dependent apoptosis) upon challenge with an oncolytic virus.

FIG. 9 shows UPR knockdown leads to ER preload in U373 glioblastoma cells as compared to GM38 normal human fibroblasts. Cells were treated with siRNA targeting IRE1α (or controls) for 72 h, total cell lysates collected at the indicated timepoints and immunoblots performed.

FIG. 10 shows UPR knockdown has no bearing on virus infectivity. (A) U373 cells were treated with siRNA (72 h) prior to Maraba infection (MOI 5). Total cell lysates were taken at the indicated timepoints post-infection and Western blots performed. (B) Cells were treated as in (A), and single-step growth analyses conducted. (C) Cells were treated as in (A), and phase contrast and fluorescent microscopy images captured following infection.

FIG. 11 shows Maraba virus infection following UPR knockdown leads to Caspase-2 activation in U373 glioblastoma cells but not GM38 normal human fibroblasts. Cells were treated with siRNA targeting IRE1α (or controls) for 72 h, total cell lysates collected at the indicated timepoints and immunoblots performed.

FIG. 12 shows a pharmacokinetic analysis of Compound 2 in CD1 nude mice. (A) CD-1 nude mice (n=3/group) were given a single dose of Compound 2 (500, 125, or 25 mg/kg, IP) and blood was drawn from the saphenous vein at 30 or 120 min. The plasma [Compound 2] was determined by LC-MS, and regression analysis conducted. (B) CD-1 nude mice (n=3/group) were given a single dose of Compound 2 (50 mg/kg, IV or 250 mg/kg, IP) and blood was drawn from the saphenous vein at 0, 5, 15, 30, 60, 120, 240, 360, 480, and 1440 min. Plasma was analyzed for [Compound 2] by LC-MS. The dotted line indicates [20 μM], which was the lowest, maximally effective dose in cell culture experiments. (C) Pharmacokinetic parameters, determined from data in (B), depicting maximum concentrations (Cmax), time at maximum concentration (Tmax), half life T_1/2), area under the curves (AUC), clearance rate (CL), volume of distribution (Vc), and relative bioavailability (F=(AUC PO/AUC IV)*100).

FIG. 13 shows chemical inhibition of IRE1α potentiates oncolytic therapy in vivo. (A) Luciferase-tagged OVCAR-4 cells (5e⁶) were delivered intraperitoneally (IP) into CD-1 nude mice. At day 14, mice were treated twice daily with Compound 2 (250 mg/Kg; IP delivery) or vehicle for 6 consecutive days (drug TX window is outlined by the dotted lines). Maraba-DM treatment (1e⁵ PFU; IV injections) was initiated 48-72 h later (virus injections depicted by arrows). Tumours were regularly evaluated using IVIS bioluminescent imaging. The graph depicts relative change in luminescent signal, which corresponds to tumour size. (B) Representative bioluminescent images from (A). (C) EMT6 cells were treated with Compound 2 (50 uM) or vehicle for 48 h prior to Maraba virus infection. Viability assays were conducted 48 h later. (D) Luciferase-tagged EMT6 cells (1e⁵) were implanted into the breast fat pads of Balb/c mice. Compound 2 treatment was initiated at Day 7 (250 mg/Kg; IP; twice daily for 6 days; treatment window depicted by the dotted box). Maraba virus injections (1e⁷ PFU; IV) commenced on Day 10 (black arrows). Bioluminescent data is plotted (as above). (E) Kaplan-meier curve depicting mouse survival in an EMT6 model. The experiment was done as in (D), except that Compound 2 treatment was extended for an additional six days. (*=p<0.05).

DETAILED DESCRIPTION

Disclosed herein are improved compositions and methods for augmenting activity of oncolytic viruses, which are obtained through the manipulation of the endoplasmic reticulum (ER) stress response. As will be described in greater detail in the following, modulators of the ER stress response pathway can sensitize tumour or cancer cells to the cytolytic activity of an oncolytic virus.

A wide range of viruses are contemplated as oncolytic viruses in the present invention, such as but not limited to herpes viruses, Adenovirus, Adeno-associated virus, influenza virus, reovirus, rhabdoviruses such as vesicular stomatitis virus (VSV) and Maraba virus, Newcastle virus, vaccinia virus, poliovirus, measles virus, mumps virus, sindbis virus (SIN) and sendai virus (SV).

Oncolytic viruses may additionally or alternatively be targeted to specific tissues or tumor tissues. This can be achieved for example through transcriptional targeting of viral genes (e.g. WO 96/39841) or through modification of viral proteins that are involved in the cellular binding and uptake mechanisms during the infection process (e.g. WO 2004033639 or WO 2003068809).

Experiments

Experiment 1:

ER Stress Response

Increased levels of unfolded proteins in the endoplasmic reticulum (ER) of all eukaryotes trigger the unfolded protein response (UPR). Several cellular pathways are involved in mitigating this stress. The ER stress pathway is responsible for dealing with unfolded protein load within the endoplasmic reticulum (reviewed in Kincaid et al., 2007, Antioxid Redox Signal, 9(12):2373-87).

Yeast have a single response to dealing with unfolded proteins through a protein kinase called IRE1. This protein kinase is activated in response to accumulated unfolded proteins within the ER and through its endoribonuclease activity, catalyses the noncanonical splicing of xbp1 mRNA to code for a functional transcription factor upregulating the expression of genes required to ameliorate the stress.

Mammalian cells also make use of the archetypal IRE1 signalling cascade in response to ER stress, but have evolved another parallel response through the ATF6 transcription factor. In unstressed cells, ATF6 resides in a transmembrane protein spanning the ER membrane. Following a stimulation of unfolded proteins in the lumen of the ER, a key at six trends locates that Golgi apparatus whereupon it is cleaved by a resident peptidase forming a soluble transcription factor responsible for the up-regulation of a subset of stress response genes. In addition to this pathway mammalian cells also possess a translational attenuation response. When unfolded proteins accumulate in the lumen of the ER, a protein kinase called PERK phosphorylates elF2alpha in the cytoplasm and attenuates global translation. This response serves to decrease ER load by stemming the influx of proteins into the ER by blocking protein production. A final response to ER stress is the selective translocation of terminally unfolded proteins from the ER into the cytoplasm for proteosome-mediated degradation. This response is termed the ER-associated Degradation (ERAD) response.

Cancer and ER Stress

Mounting evidence demonstrates that during the etiology of a tumour, cancer cells undergo sustained and/or transient ER stress. Mutations give rise to protein species with suboptimal folding, and hypoxia impedes proper folding of proteins within the ER. Several components of the ER stress response pathway are upregulated in a variety of cancers (Shuda et al., 2003, J. Hepatol. 2003, 38(5):605-14). Manipulating ER stress sensitizes cancer cells to hypoxia (Bi et al., 2005, EMBO J., 24:3470-81) and to chemotherapeutics (Nawrocki et al., 2005, Cancer Res. 65:11658-66).

Viruses and ER Stress

Viruses induce ER stress, for example: HCV (Joyce et al., 2009, PLoS Pathog., 5(2):e1000291), SARS (Ye et al., 2008, Biochim Biophys Acta.,1780:1383-7), West Nile (Medigeshi et al., 2007, J. Virol., 81:10849-60), Hepatitis B (Li et al., 2007, Virus Res., 124:44-9), Hantavirus, Japanese Encephalitis virus (Su et al., 2002, J. Virol., 76:4162-71), RSV (Bitko et al., 2001, J. Cell Biochem., 80:441-54), influenza (Watowich et al., 1991, J. Virol., 65:3590-7), Herpes (Lee 2008), and dengue fever virus (Umareddy et al., 2007, Virol J., 4:91). Some viruses have the ability to manipulate the cell's response to ER stress: (Bechill et al., 2008, J. Virol.; 82:4492-501; Isler et al., 2005, J. Virol., 79:6890-9; Tardif et al., 2004, J. Biol Chem., 279:17158-64).

Replicating virus based therapeutics, or oncolytic viruses, are a rapidly emerging and promising treatment modality for a wide range of cancers. In pre-clinical studies, oncolytic viruses have produced remarkable results in a variety of experimental animal models including human xenografts in nude mice and syngeneic animal tumours (see Hawkins et al., 2002, Lancet Oncol. 3:17-26; and VähäKoskela 2007 for review). Successfully tested oncolytic viruses include: vesicular stomatitis virus (VSV) (Stojdl et al., 2003, Cancer Cell., 4:263-75; Lun et al., 2006, J. Natl. Cancer Inst., 98:1546-57), adenovirus (AdV) (Ries et al., 2002, Br. J. Cancer, 86:5-11), reovirus (Coffey et al., 1998, Science, 282:1332-4), Newcastle disease virus (NDV) (Lorene et al., 1994, J. Natl. Cancer Inst., 86:1228-33; and Schirrmacher et al., 2001, Int. J. Oncol., 18:945-52), herpes simplex virus (HSV) (Todo et al., 1999, Hum. Gene Ther., 10:2741-55) and vaccinia virus (McCart et al., 2001, Cancer Res., 61:8751-7). Several of these viruses have been, and are continuing to be tested in human clinical trials; again with encouraging results. For example, in a phase I trial, using a genetically modified herpes virus (HSV G207), patients with malignant gliomas were injected intratumourally and some antitumour efficacy was seen by both radiographic and neuropathologic criteria (Markert et al., 2000, Gene Ther.,7:867-74). Onyx-015, an E1B-55kDa gene-deleted adenovirus has completed two phase II human trials directed at squamous cell carcinomas of the head and neck (Nemunaitis et al., 2001, J. Clin. Oncol., 19:289-98; Lamont et al., 2000, Ann. Surg. Oncol., 2000, 7:588-92). In these studies virus was delivered by a series of intra-tumour injections either in combination with chemotherapy agents (Lamont et al., 2000, Ann. Surg. Oncol., 2000, 7:588-92) or as a single agent (Nemunaitis et al., 2001, J. Clin. Oncol., 19:289-98). Onyx-015 was found to be safe and showed some antitumour activity with 10-30% of patients showing complete responses at injection sites and 30-60% of patients having stabilized disease. In a more recent phase I trial, an engineered form of vaccinia virus demonstrated an excellent safety profile as well as promising efficacy data in 14 patients (Park et al., 2008, Lancet Oncol., 9:533-42). There have been in excess of some 25 clinical trials (mostly phase I) that demonstrate the safety of these virus therapies. Results from more phase 2 and 3 trials are awaited to evaluate the efficacy of these oncolytic viruses.

Rhabdoviridae:

The Rhabdoviridae viral family is divided into 6 genera, in which the vesicular stomatitis virus (VSV) is one of them. Rhabdoviridae are membrane-enveloped viruses that are widely distributed in nature where they infect vertebrates, invertebrates, and plants. Viral particles contain a helical, nucleocapsid core composed of genomic RNA and protein. Rhabdoviridae have single, negative-strand RNA genomes of 11-12,000 nucleotides. Further information on the Rhabdoviridae family of viruses can be found in Rose and Whitt, 2001, Chapter 38, Rhabdoviridae: The viruses and their replication, in Fields Virology, 4.sup.th edition, pp. 1221-1244, the entirety of which is hereby incorporated by reference.

The inventors have previously shown that VSV has oncolytic properties (Stojdl et al., 2000, Nat. Med., 6:821-5) and have since shown that the VSV M protein antagonizes the innate immune system by blocking nuclear cytoplasmic transport of host mRNA. In doing so, the transcriptional cascade responsible for perpetuating the interferon mediated antiviral program is severed and no IFN is produced from these infected cells (Stojdl et al., 2003, Cancer Cell., 4:263-75). VSV strains with M protein mutations lose their capacity to block the IFN response and were shown to be extremely attenuated in normal cells, yet retain their ability to kill tumour cells (Stojdl et al., 2000, supra; Stojdl et al, 2003, supra). In a variety of subcutaneous, metastatic lung and intraperitoneal mouse models of cancer, systemic injection of the engineered VSV mutants was shown to effectively cure mice of local and disseminated tumours (Stojdl et al., 2000, supra; Stojdl et al. 2003 supra).

Oncolytic virus strains from the rhabdovirus family are described in WO 2009/016433, which is herein incorporated by reference.

RNAi Technology:

The recent advent of RNAi technology has made it possible to use forward genetics techniques to study the function of mammalian genes (Berns et al., 2004, Nature, 428:431-7; Krishnan et al., 2008, Nature, 455:242-5). This technology is particularly useful for studying host virus interactions as many of the host systems relevant to virus infection are unique to higher order organisms (e.g. interferon signaling).

The present inventors have utilized a genome wide RNAi screen to identify host genes, which when neutralized, sensitize cells to a subsequent oncolytic virus infection resulting in increased cell death (FIGS. 1 and 2). This sensitization is specific to cancer cells and does not sensitize normal human primary fibroblasts to oncolytic virus infection (FIG. 3). Accordingly, a means for specifically sensitizing cancer cells to killing by oncolytic virus-based therapy is provided.

Table 1 lists components of the endoplasmic reticulum (ER) stress pathway that, when removed from the cell or deactivated according to an embodiment of the invention, makes the cell more susceptible to killing by a subsequent infection with an oncolytic virus, for example rhabdovirus-based oncolytic viruses. This is demonstrated by in vitro cytotoxicity assays across a panel of cancer cells using a panel of oncolytic agents as shown in FIGS. 3 and 4.

TABLE 1
Components of the UPR and ERAD pathways identified as synthetic lethal with Maraba
virus infection in both OVCAR 8 and U373 human cancer
cells.
UPR	ERAD
Symbol	GeneID	RefSeq	Synonym	Symbol	GeneID	RefSeq	Synonym
ATF6	22926	NM_007348.2		DERL1	79139	NM_024295.4
CREBL1	1388	NM_004381.4	atf6 beta	DERL2	51009	NM_016041.3
ERN1	2081	NM_001433.3	IRE1alpha	sec61a	29927	NM_013336.3
NFYC	4802	NM_014223.4		sec61g	23480	NM_014302.3
HSPA5BP	54972	NM_178031		Dnajb9	27362	NM_013760.4	erdj4
FKBP10	60681	NM_021939		DNAJB11	51726	NM_016306.4	Erdj3
SEP15	9403	NM_004261		AMFR	267	NM_001144.4

Without wishing to be bound by theory, the enhanced tumour killing capacity is proposed to improve oncolytic virus efficacy by increasing tumour cell death following infection by an oncolytic virus and thereby debulking the tumour more rapidly and requiring less oncolytic virus at the tumour site to achieve similar efficacy.

Tumours are variably and intermittently hypoxic. This is because the vasculature that feeds tumours is often poorly structured. Hypoxia induces ER stress in a number of ways (reviewed in Wouters 2008, supra). It has been proposed that blocking UPR mechanisms would sensitize hypoxic tumour cells to death due to their dependence on these rescue pathways. However, some portions of tumours (often the rims) are not hypoxic as they are fed oxygen from the surrounding healthy stroma, or are adjacent to properly functioning vasculature. These areas of tumours that are not themselves hypoxic would not be affected by ER stress response blocking agents. However, an oncolyic virus infection of these non-hypoxic tumour cells would kill these cells efficiently in combination with ER stress response blockade. Since only tumour cells will be infected by the oncolytic virus, we refer to this as “targeted ER stress”. This combination of oncolytic virus and ER stress response blockade would result in a more complete tumour cell ablation and lessen the chance of re-growth of the tumour; a common problem with current chemotherapy.

Alternatively, it has been shown that oncolytic therapy can induce vascular shutdown and catastrophic hypoxia within tumour cores (Breitbach et al., 2007, Mol. Ther., 15:1686-93). Combination therapy of oncolytic virus with ER stress response blockade would be promoted by the hypoxia induced by the oncolytic virus, even in distant cells not directly infected by the virus. This would again limit the probability that a tumour cell would escape treatment and thereby improve patient outcomes.

Chemical signals (chemokines/cytokines) from the infected cell are released to warn neighboring cells of an imminent virus infection. For example, interferon beta is released from infected cells and induces a paracrine and autocrine signaling cascade that results in a potent antiviral response. Some tumour cells are capable of responding to these chemical signals and mount a defense against an incoming oncolytic virus. It has been demonstrated that interferon type I receptor is downregulated during ER stress. Without wishing to be bound by theory, we propose that inducing an ER stress following oncolytic virus infection will decrease the ability of the infected cell to secrete chemokines/cytokines and as well as the receptors that are required to sense these chemical signals. Further inhibiting of the ER stress response, through combination therapy with a drug or by engineering the oncolytic virus to block this response, should additionally attenuate the chemokine/cytokine mediated antiviral defenses of the infected cell and the surrounding tumour cells. Since the normal healthy cells are themselves resistant to the oncolytic agents, they would not be significantly affected by this mechanism. Therefore, ER stress modulation of innate immunity would specifically sensitize tumour cells to oncolytic virus infection.

Experimental Design:

A genome wide RNAi screen was conducted to find host genes that could modulate the ability of an oncolytic virus to kill tumour cells. Maraba virus was selected as a representative oncolytic virus from the Rhabdoviridae family. In the following experiments human cancer cells were sensitized to Maraba virus infection by interfering with host cell mRNA expression using siRNA technology. Two cell lines: (1) OVCAR 8 human ovarian carcinoma cells; and (2) U373 human glioblastoma cells, were studied as representative unrelated malignancies in an effort to identify genes that were common to many cancers, and not necessarily specific to one indication. Genes were identified that, when inhibited or augmented, gave rise to improvements in oncolytic activity.

“Hits” from the screens were analyzed for their known functions and it was determined that several of these genes were components of the host ER stress response pathways. Specifically, IRE1 and ATF6alpha and ATF6beta were identified as components of the UPR. IRE1 is known to activate the transcription factor XBP1 through a non-canonical mRNA splicing mechanism in the cytoplasm. Interestingly, the transcriptional co-activator NFYC known to bind and cooperate with both ATF6 and XBP1 was also identified as a “hit”. In addition, components of the ER activated Degradation (ERAD) pathway were identified in the primary screen. AMFR and DERL are known to form a complex and are responsible for tagging and removing terminally unfolded protein from the ER for proteosomal degradation.

These results strongly indicated that modulating ER stress responses through multiple pathways all sensitized cells to Maraba virus infection induced cell death.

Experimental Procedures:

Genome-wide Screening Procedure: Cells were reverse transfected in 384 well format using 10 nM of Dharmacon siGenome SmartPool human set (Invitrogen USA). For OVCAR 8 human ovarian carcinoma cells 2500 cells/well were transfected using RNAiMax (Invitrogen, USA) (0.05 ul/well) in a total volume of 40 ul of DMEM containing 10% FBS. Alternatively U373 human glioblastoma cells were similarly reverse transfected using Oligofectamine (0.05 ul/well) at a density of 625 cell/well. Plates were incubated for 72 hours to allow for siRNA mediated mRNA down modulation at which time plates were either mock infected or infected with recombinant wild type Maraba virus at an MOI of 0.05. To assay for cell death, plates were incubated for a further 48 hours and then resazurin was added to a concentration of bug/ml. After 4 hours, absorbance readings at 605 nm and 575 nm were taken to monitor reduction of resazurin to resorufin as a measure of cell viability (O'Brien et al., 2000, Eur J Biochem., 267:5421-6). All screens were performed in duplicate.

Data Analysis: Viability scores for each well were normalized using negative controls (irrelevant siRNA transfection) on a per plate basis. Duplicate screens were averaged on a per well basis. The mean standard deviation for all negative control wells was calculated and used to represent the variability in the assay. Experimental wells which deviated from the mean of all experimental wells by a value equal to 2 standard deviations of the negative controls from their mean were scored as “meaningful”. The mock-infected version of the screen (siRNA alone) was used to remove false positive “hits”. Data was normalized as above. Experimental wells which deviated from the mean of all experimental wells by a value equal to 1 standard deviation of the negative controls from their mean were scored as “meaningful”. Gene targets were designated as “hits” if they were only meaningful in the virus infected (and not in the virus uninfected) screens.

Validation experiments: Several experiments were performed to validate the “hits” identified in the primary siRNA screen. Firstly, we wanted to determine if Maraba virus infection could induce a UPR response. By definition, the “hits” derived from the screen were dependent on virus infection. Therefore, we predicted that the virus must be inducing an ER stress which was not present during the siRNA alone control arm of the screen. There are three arms to the UPR response: (1) ATF6 (2) IRE1 and (3) PERK. Each is known to sense unfolded protein load within the ER by a BIP dependent mechanism. We infected U373 and OVCAR8 cells with Maraba virus and assayed the kinetics of the 3 arms of the UPR response (FIG. 4 panel A&B). As expected, virus infection led to robust ATF6 cleavage indicative of ATF activation during ER stress and persisted throughout the infection (FIG. 4A). IRE1 activation was observed at 4 hours post infection and had subsided 24 hrs post stress. eIF2 alpha is phosphorylated by several protein kinases in response to a variety of stresses (FIG. 4 B). Tunicamycin is seen inducing elF2alpha phosphorylation presumably by the protein kinase PERK (FIG. 4A). We see a robust induction of elF2alpha phosphorylation following Maraba virus infection and we presume that at least some of this activity is through the PERK kinase. Thus Maraba virus infection does indeed elicit an ER stress response. Finally we sought to validate IRE1 as an ER stress response gene whose down-modulation would result in sensitization to the ER stress induced by Maraba virus infection. We targeted IRE1alpha (also known as ERN1) and IRE1beta (also known as ERN2) using siRNA from another vendor (Qiagen USA) across a panel of human cancer cell lines. Subsequent infection with increasing amounts of virus determined the dose response curve for each cell line. FIG. 3C demonstrates the efficiency of siRNA knockdown using this methodology. From this experiment, it was determined that the optimal siRNA concentration for the validation experiments would be 10 nM. FIG. 3A shows a typical curve demonstrating sensitization of a human glioblastoma cell line (SF295) to Maraba infection by knocking down either IRE1 alpha or IRE1beta relative to an irrelevant siRNA control. The summary of the experiment is presented in FIG. 3B, showing that the vast majority of tumour lines tested are sensitized to Maraba virus infection when IRE1 is downregulated. Importantly, GM38 primary human fibroblasts were not sensitized to Maraba virus infection. This demonstrates a tumour specific sensitization when modulating ER stress responses. We propose that this tumour specific sensitization will help target Maraba oncolytic virus destruction to the tumour and spare the surrounding normal parenchyma.

Cell culture: For immunoblot and RT-PCR experiments, U373 (2×10⁵) and OVCAR8 (5×10⁵) cells were seeded in 35 mm plates and grown overnight in complete DMEM. The following morning, tunicamycin (5 μg/mL) and Maraba virus (MOI 5) were diluted in fresh DMEM and added to the cells. Cell pellets were collected at the indicated timepoints post-treatment, washed twice in cold PBS with complete protease inhibitors (Roche) and stored at −80° C. until lysis. For viability experiments, 5×10³ cells were seeded in 96-well plates and grown overnight in complete DMEM. The following morning, siRNA knock-down was performed using RNAimax reagent (Invitrogen) and chemical duplexes specific to human IRE1α (ERN1) or β (ERN2) or a non-targeting (NT) control (Qiagen). After 72 hours, log-dilutions of Maraba virus were added (in triplicate), and 48-72 hours later cell viability was analyzed using the alamar blue method.

Immunoblotting: Total cell lysis buffer (50 mM Tris-HCl; 150 mM NaCl; 1% Triton X-100; 1% SDS) was added to cell pellets, and the lysates were “sheared” using a p100 pipette tip. Total cell lysates were prepared in SDS sample buffer, and 5-50 ug of total protein was separated by SDS-PAGE on 10% Bis-Tris gels and transferred to PVDF membranes. Membranes were probed with primary antibodies diluted in 5% skim milk powder (SMP) overnight at 4° C., followed by horse radish peroxidase-conjugated secondary antibodies diluted in 5% SMP for 1 h at room temperature. Membranes were then treated with ECL reagent, exposed to X-ray film and developed (Kodak X-OMAT 2000A). Primary antibodies used were: ATF-6α (Santa Cruz), p-EIF2α (Cell Signaling), Bip/Grp78 (Cell Signaling), XBP(s) (Biolegend), GAPDH (Advanced Immunochemicals), Maraba viral proteins (anti-VSV).

RT-PCR: Cell pellets were lysed, and RNA extracted using a Qiagen RNAeasy Mini kit. RNA purity and concentration were determined spectrophometrically, and RT-PCR was performed using standard procedures with oligo-dT primers and the following XBP-specific primers:

(SEQ ID NO: 1)
For: 5′- cct tgt agt tga gaa cca gg -3';
(SEQ ID NO: 2)
Rev: 5′- ggg gct tgg tat ata tgt gg -3′.

The PCR product was separated on a 2% Nusieve/1% Agarose gel and visualized under UV.

Experiment 2:

Blockade of ER Stress Response Sensitizes Cancer Cells Towards Viral Oncolysis:

To search for host factors that modulate rhabdovirus-mediated oncolysis, a synthetic lethal RNAi screen of the human genome was performed across three tumour-derived cell lines (FIG. 5a). We used an arrayed library of siRNA pools to target ˜18 500 genes in OVCAR-8 (ovarian carcinoma), U373 (glioblastoma) or NCI-H226 (non-small cell lung carcinoma) cells. Transfected cells were either mock infected or infected with wild type Maraba virus as a representative oncolytic rhabdovirus. Following infection, we incubated the cells for 48-72 h after which we scored cell viability using resazurin vital dye. To identify primary “hits”, we analyzed data from two independent screens for each cell line using the median absolute deviation method⁹. Subtracting those genes scoring positively in the siRNA alone screens defined 1008 synthetic lethal hits common to at least two out of three cancer lines from the primary screen (Table S1). Subsequent bioinformatics analysis revealed a striking enrichment of hits within the ER stress response pathways (FIG. 1B), including members of two of the three known signaling cascades that comprise the unfolded protein response (UPR). Key hits therein included the transcription factors ATF6α and ATF6β, the endoribonucleases/protein kinase IRE1α, and a transcriptional coactivator common to both pathways, NFYC. Together, the ATF6 and IRE1 pathways serve to rescue the ER from an overload of unfolded proteins by increasing chaperone production and ER lipid biogenesis¹⁰. Our screen also identified several members of the SEC61 and the HRD ligase protein translocation complexes (e.g. Derlin-1; FIG. 1B). These proteins are critical for ER-associated degradation (ERAD), which helps rescue an unfolded protein burden by removing misfolded polypeptides from the ER and shuttling them to the 26S proteasome¹¹.

UPR and ERAD components were particularly interesting as sensitizers because ER stress has been reported to be a defining feature of the tumour cell state and components of these pathways are already being pursued as cancer specific targets for stand-alone cancer treatment^12,13. We thus performed secondary validation for several members of these pathways, using siRNA with targeting sequences distinct from those employed in the primary screens. Depletion of IRE1α, ATF6α or Derlin-1 significantly sensitized U373 glioblastoma-derived cells to virus-mediated killing across a range of multiplicity of infections (MOI; FIG. 5C-D). To rule out off-target effects, we used multiple siRNA duplexes targeting distinct sequences and consistently found that UPR knockdown sensitized towards viral oncolysis (FIG. 6). In addition, we performed a rescue experiment by ectopically expressing murine ATF6α in U373 cells before depleting its human counterpart. Cells stably expressing mATF6α were completely refractory to the synthetic lethal phenotype associated with oncolytic virus infection and hATF6α knockdown (FIG. 5D). Taken together, these experiments validate our primary screening result that rhabdovirus-mediated oncolysis is greatly enhanced by knocking down components of the UPR and ERAD pathways.

To evaluate therapeutic index, we silenced IRE1α in a small panel of primary human cell lines (GM38 skin fibroblasts, normal human astrocytes (NHA) and Wi38 lung fibroblasts) prior to rhabdovirus infection. In contrast to the pronounced sensitization observed in U373 glioblastoma cells, UPR inhibition did not alter Maraba virus-mediated killing of the normal cell lines (FIG. 5E). We next examined the scope of the synthetic lethal phenotype in a representative subset of the NCI 60 tumour cell panel. RNAi-mediated knockdown of IRE1α or ATF6α significantly sensitized >80% of cancer cell lines tested to virus-mediated killing (FIG. 5F). The sensitized cell lines represent a broad assortment of cancers, some of which had fully effective interferon systems while others had varying degrees of interferon defects (FIG. 5F). Oncolytic rhabdoviruses traditionally have difficulty killing tumour cells with intact interferon responses^7,8. Blockade of ER stress responses appears to extend the capability of these viruses to kill such cells, which may result in greater efficacy in the clinical setting where tumours are expected to be more heterogeneous with regards to interferon signaling. Oncolysis by the prototypic oncolytic rhabdovirus VSV and an engineered clinical candidate strain of Maraba virus (Maraba-DM)⁶ was similarly enhanced by UPR inhibition (FIG. 5G). Collectively, these data suggest that the enhancement of virus-mediated oncolysis conferred by inhibiting the ER stress response is tumour cell specific and may have widespread utility across a diverse range of tumour types.

Synthetic Lethal Interaction Between ER Stress Response Blockade and Rhabdovirus Infection Requires a Preconditioning Process

Maraba virus infection caused noticeable ER stress characterized by the activation of the upstream UPR sensors IRE1α (measured by XBP1 mRNA splicing (FIG. 7A)), ATF6α (measured by its cleavage (FIG. 7B)) and PERK (measured by EIF2α phosphorylation (FIG. 7B)). Surprisingly, however, in spite of the activation of these stress sensors, representative downstream UPR effector proteins XBP1(s) and BiP were not elevated after virus infection (FIG. 7B). This result indicates that the UPR was stalled at an early stage during virus infection and thus rendered functionally inert. These data suggest that an inadequate ER stress response is unlikely to be responsible for the observed synthetic lethal interaction between virus and UPR/ERAD knockdown, as the UPR is inhibited rapidly upon virus infection independent of external manipulation. Instead, it appears that sustained inhibition over the 72 h knockdown period is required, and that this may precondition cancer cells to die in response to virus infection.

To test this idea directly, we synthesized a number of compounds that had been reported to inhibit IRE1α¹⁶, along with some novel variants of the original structure. We first tested these compounds for their ability to inhibit XBP1 splicing by IRE1α and found that several were effective in the micromolar range (representative subset depicted in FIG. 7C). We then evaluated the most potent of these, designated Compound 2, and found that it greatly enhanced viral oncolysis in U373 cells when dosed for 48 hours, but not 4 hours (FIG. 7D). Importantly, combination index analyses demonstrated that compound 2 interacted synergistically (CI<1.0) with Maraba virus across a range of doses and MOIs (FIG. 7E). We next treated U373 cells with cyclosporine, a potent inhibitor of the ER chaperone protein cyclophilin B, prior to virus infection. Similar to compound 2, cyclosporine pre-treatment for 48 h but not 4 h greatly enhanced viral oncolysis (FIG. 7F). Together, these drug data demonstrate that the synthetic lethal phenotype is due to a preconditioning process that occurs throughout a period of sustained inhibition of the ER stress response, as opposed to acute blockade.

ER Preload Rewires Cancer Cells for Apoptosis

We examined whether inhibiting the ER stress response induces an unfolded protein load prior to viral infection (i.e. “ER preload”). Because there are presently no direct measures of ER protein load per se, we measured activation of the UPR as an indirect readout. Here, IRE1α silencing led to a tumour cell-specific increase in the ER stress-responsive proteins BiP and Mcl-1, as well as a transient induction of the ER stress sensor PERK (as measured by P-EIF2α; FIG. 8A and FIG. 9). These data indicate that IRE1α inhibition resulted in an ER stress in tumour cells. However, this stress response appeared to resolve as eIF2α phosphorylation and ATF6 levels returned to untreated levels, before a period at which virus infection was to be initiated (FIG. 8A 72 h and FIG. 8B 0 h). Additionally we examined whether ER preload altered the kinetics of the ER stress response post-virus infection and observed that loss of IRE1α had no bearing on ATF6α cleavage, EIF2α phosphorylation and Mcl-1 turnover (FIG. 8B), also consistent with our findings that the ER was not under duress at the time of infection. Collectively, these data indicate that inhibiting the ER stress response in tumour cells leads to a transient stress response we refer to here as “ER preload”. We thus asked whether ER preload is accountable for preconditioning cells to respond differently than naïve cells to subsequent oncolytic virus infection. Thus we chemically induced an ER preload by pulsing cells with the glycosylation inhibitor tunicamycin 48 hours before virus infection (FIG. 8C), and found this to also enhance virus-mediated killing. These data are consistent with an ER preload, induced either upon sustained UPR inhibition or ER poisoning, to be requisite for the synthetic lethal phenotype with virus infection.

We wished to identify the mechanism of how ER preload might synergize with a subsequent virus infection to promote tumour cell death. We noted that UPR inhibition had no bearing on viral protein expression (FIG. 8B), and confirmed that it also did not alter any aspect of the virus life cycle (FIG. 10). We thus hypothesized that ER preload might “rewire” cancer cell signaling to initiate apoptosis when subsequently challenged with an oncolytic virus. Indeed, IRE1α knockdown greatly enhanced the kinetics of apoptosis during a viral infection, as measured by the cleavage of PARP as well as members of the caspase cascade (FIG. 8D). Notably, caspase 2 was strongly activated in tumour cells by virus infection only when IRE1α was knocked down (FIG. 8D and FIG. 11). Caspase 2 is an initiator caspase that has been implicated in several stress-mediated apoptotic cascades, such as those emanating from DNA damage¹⁸ as well as ER stress¹⁹. It has been reported that this caspase 2 initiated death pathway remains dormant until unresolved ER stress triggers its activation¹⁹. To examine the relevance of its activation, we knocked it down simultaneously with IRE1α and measured apoptosis following virus infection. Remarkably, caspase 2 knockdown largely rescued the synthetic lethal interaction between IRE1α knockdown and virus infection (FIG. 8E). Together, these data suggest that transient ER preload rewires cancer cells to undergo caspase-2 dependent apoptosis upon virus infection (FIG. 8F).

Chemical Inhibition of IREα Enhances Viral Oncolysis In Vivo

We sought to evaluate the efficacy of pharmacologic ER stress response blockade combined with oncolytic virus therapy in animal models of cancer. To begin, we undertook maximum tolerable dose (MTD) and pharmacokinetic (PK) studies of Compound 2 in CD-1 nude mice. These experiments showed that a single dose of up to 1000 mg/Kg of Compound 2 was tolerated, had a half-life of >6 hours, and had properties consistent with efficient bio-distribution to the extravascular tissues (FIG. 12).

As with many cancers, ovarian carcinoma is difficult to treat clinically due to development of resistance to current therapies²⁰. Thus, to validate our combination therapy approach, we chose a chemoresistant, orthotopic OVCAR-4 xenograft model²¹ that is also refractory to oncolytic virus therapy. OVCAR-4 cells stably expressing firefly luciferase were injected intraperitoneally (IP) into CD-1 nude mice. We monitored tumour growth using in vivo optical imaging, and initiated treatment during the growth phase. To induce ER preconditioning, we treated animals with Compound 2 for three days prior to the first virus injection. Consistent with our findings in cell culture, combination therapy dramatically reduced tumour burden in animal models, an effect that was sustained for >30 days with negligible tumour re-growth (FIG. 13A-B). In contrast, rapid re-growth occurred after an early period of tumour regression using either virus or Compound 2 alone.

As a complement to these experiments in human xenografts, we sought to test this treatment regiment in an immune competent rodent tumour model. In vitro testing determined that the EMT6 breast cancer line, which is particularly resistant to stand-alone rhabdovirus therapy, was significantly sensitized to oncolytic virus killing when pre-treated with Compound 2 (FIG. 13C). Using these cells to generate a tumour model, we confirmed that neither drug nor virus had an appreciable effect on tumour growth as single agents; however, combination therapy significantly reduced tumour burden (FIG. 13D). Notably, when drug treatment was stopped, tumour re-growth occurred even in the presence of continued virus dosing, validating the inter-dependence of the treatment combination. Extending Compound 2 treatment to more than 12 days in combination with virus treatment continued to increase efficacy (FIG. 13E). Taken together, these data demonstrate proof of concept that modulating the ER stress responses can be exploited to enhance oncolytic virus therapy in vivo.

Materials and Methods:

Cell culturing: Human 293T (American Type Tissue Collection (ATCC)), Monkey Vero (ATCC), murine EMT6 (ATCC), human GM38 (National Institute of General Medical Sciences Mutant Cell Repository, Camden, N.J.), human Wi38 primary fibroblast (ATCC) and cell lines from the NCI 60 cell panel (obtained from the Developmental Therapeutics Program, National Cancer Institute (Bethesda, Md.)) were propagated in Dulbecco's modified Eagle's medium (Hyclone, Logan, Utah) supplemented with 10% fetal calf serum (Cansera, Etobicoke, Ontario, Canada) using standard tissue culture procedures. Normal human astrocytes were propagated in astrocyte media (Sciencecell Research Laboratories) and cultured using standard procedures.

Virus production: Vero cells were plated in 15 cm format, grown to confluence (−2.5×10⁷) and infected with Maraba-WT, Maraba-DM or VSV-WT viruses at MOI 0.1. After 18 h, the virus- containing cell culture media was collected and centrifuged at ˜18,600×g for 1.5 h. The virus pellet was carefully washed and re-suspended in PBS (10 mL), and gently over-layed onto a 20% sucrose solution (1 mL). After ultracentrifugation for 1.5 h (26,900 rpm), the pellet was re-suspended in 15% glucose, aliquoted and stored at −80 deg C.

RNAi screening: An arrayed library of siRNA pools (Dharmacon, Thermo Fisher, USA) was used to target ˜18,500 human genes in OVCAR-8 (ovarian carcinoma), U373 (glioblastoma) or NCI-H226 (non-small cell lung carcinoma) cells. Tumour cells were seeded in 384 well plates (OVCAR-8=1250 cells/well, U373=625 cells/well, NCI-H226=625 cells/well) and allowed to grow for 24 h. Each plate had additional control wells with a non-targeting control siRNA (Dharmacon non-targeting Pool #2) to measure the effect of siRNA transfection on infection, and siRNA targeting PLK-I (Dharmacon) was used to monitor knockdown efficiency. Quadruplicate plate sets were reverse transfected with siRNA (10 nM) using RNAimax (Invitrogen, USA) and incubated for 72 h. From these, duplicate sets of plates were either mock infected or infected with wild type Maraba virus (MOI: OVCAR-8=0.1, U373=0.5, NCI-H226=0.1). Infections were incubated for 48 h (OVCAR-8) or 72 h (U373 and NCI-H226) after which resazurin dye (20 μg/mL) was added to each well, incubated for 6 h and assayed for absorbance (573 nm) to score cell viability.

Data Analysis: Cell viability data from the screens was normalized on a per plate basis using the Median Absolute Deviation (MAD) method (1). Briefly, for each well on the plate, an absolute deviation from the plate median (WAD) was calculated using the formula:

WAD=(well value−plate median excluding controls)

A MAD was calculated for each plate using the formula:

Plate MAD=1.4826* median (WAD)

A MAD score for each gene target (gMAD Score) was calculated as follows:

gMAD Score=average of 2 euplicates (WAD/Plate MAD)

A composite gMAD score for each gene target was derived by subtracting the gMAD scores from the mock-infected screens from infected screens, for each cell line. Gene targets scoring less than −1.85 were considered synthetic lethal hits. Hit lists were derived for each cell line and then compared using VENNY (2) to obtain a final list of hits identified in at least 2 out of the 3 cell lines screened (1008 hits; Table 2). Bioinformatics analysis of the composite hit list was performed using a combination of PANTHER (3), DAVID (4), Ingenuity Pathway Analysis (Ingenuity Systems, USA), and manually curated to identify signaling pathways enriched with hits and to annotate hits for gene function and sub-cellular localization.

RNAi reagents for secondary screening: For all RNAi experiments, the following mRNA sequences were targeted with chemically-synthesized duplexes: IRE1α, 5′-cag cac gga cgt caa gtt tga-3′ (Qiagen) (SEQ ID NO: 3); ATF6α, 5′-cag caa cca att atc agt tta-3′ (Qiagen) (SEQ ID NO: 4); Derlin-1, 5′-tcc cgg cga tca cgc gct att ggt t-3′ (Invitrogen) (SEQ ID NO: 5); Caspase 2 (Dharmacon Smart Pool); Non-targeting (NT) #1, 5′-gca cca tgc ctt tga gct t-3′ (Invitrogen) (SEQ ID NO: 6); NT #2 (Dharmacon NT pool #1). For experiments in FIG. 6, the following sequences were targeted: IRE1α, 5′-ccc tac cta cac ggt gga cat ctt t-3′ (Invitrogen #615) (SEQ ID NO: 7); IRE1α, 5′-gac ctg cgt aaa ttc agg acc tat a-3′ (Invitrogen #847) (SEQ ID NO: 8). All RNAi transfections were performed using RNAimax (Lipofectamine) and left for 72 h before further manipulation. Experiments were done using a [siRNA]=10 nM, except caspase 2 RNAi experiments, which were done at 50 nM.

In vitro cytotoxicity assays with RNAi: Cells were seeded onto 96 well plates to a confluence of ˜50%. The following day, siRNA transfections were performed, and 72 h later the cells were infected at log 10 dilutions with wild type Maraba virus (except for FIG. 5G, which used the indicated viruses). After 48-72 h of infection (depending on the cell line), Resazurin sodium salt (Sigma-Aldrich) was added to a final concentration of 20 μg/ml. After a six-hour incubation, the absorbance was read at a wavelength of 573 nm. To determine “log sensitization”, kill curves were plotted on a log 10/linear graph and EC50 values determined. The log sensitization was calculated by subtracting the EC50 of UPR targeted from non-targeted cell lines, and is represented as log 10 values.

Lentiviral production and rescue experiments: Total RNA was extracted from C2C12 myoblasts using RNeasy technology (Qiagen), and reverse transcribed using random hexamers and Superscript II (Invitrogen). The following primers were used to PCR amplify mouse ATF6α from this cDNA library: Forward, 5′-ggt acc gcg ggc gcg cca tgg agt cgc ctt tta ctc cgg-3′ (SEQ ID NO: 9); Reverse, 5′-ctt gga tcc gcg gcc tac tgc aac gac tca ggg atg-3′ (SEQ ID NO: 10). PCR amplicons were cloned into a pLEX lentiviral vector (Open Bioststems) using the In-Fusion Advantage PCR cloning kit (Clontech). Lentivirus particles were produced by reverse transfecting pDY-ATF6α, pCMV 8.74, and pMD2-G vectors (Fugene-6 transfection reagent, Roche) into 293-T cells. After 72 h, the virus-containing media was removed, passed through a 0.45 μM filter, aliquoted, and frozen at −80 deg C. For rescue experiments, U373 cells were seeded in 6-well format to ˜30% confluence. The following day, lentiviral-containing media was diluted 1:1 with complete media and polybrene was added to a final concentration of 6 μg/mL. Diluted media was added to cells, and plates were spun at 400×g for 1 hour. The following day, siRNA transfections were performed and 72 h later Maraba-WT virus was used to infect the cells. A Resazurin sodium salt cytotoxicity assay was performed 48 h post-infection.

Immunoblotting: Cells were lysed (50 mM Tris-HCl; 150 mM NaCl; 1% Triton X-100; 1% SDS) and protein quantified using the Lowry assay (Bio-Rad). Total cell lysates were prepared in SDS sample buffer, and 5-50 μg of total protein was separated by SDS-PAGE on Bis- Tris gels (ranging from 8-15%) and transferred to nitrocellulose or PVDF membranes. Membranes were probed with primary antibodies diluted in 5% skim milk powder (SMP) or 5% Bovine Serum Albumen (BSA) overnight at 4 deg C., followed by horse radish peroxidase-conjugated secondary antibodies diluted in 5% SMP for 1 h at room temperature. The following primary antibodies were used: rabbit mAb anti-IRE1α (Cell Signaling 14C10); rabbit anti-ATF-6α (Santa Cruz Biotechnology, H-280); rabbit anti-Derlin-1 (Sigma); mouse anti-GAPDH (R&D Systems); rabbit anti-XBP1 (BioLegend, Poly6195); rabbit anti-BIP (Cell signaling); rabbit anti-phospho-EIF2α (Cell signaling); rabbit anti-VSV; rabbit anti-Mcl-1 (Santa Cruz Biotechnology, S-19); goat anti-human IFN cup α/β R1 (R&D S_ystems); rat anti-Caspase-2 (Chemicon, 11B4); rabbit anti-Caspase-3 (Cell Signaling, Asp175); rabbit anti-Caspase-9 (Cell Signaling, human-specific); rabbit anti-PARP (Cell Signaling). Finally, proteins were visualized using SuperSignal West Pico Chemiluminescent Substrate System (Pierce Biotechnology).

Interferon production assay: An indirect “interferon production bioassay” was used to estimate the degree to which our cell lines could produce interferon. The indicated lines were infected with Maraba-Δ51 (MOI 3) to trigger an innate immune response and induce interferon (IFN) production. Eighteen hours later, the interferon-containing media was collected and acid neutralized with 0.25N HCl overnight at 4° C. (to destroy virus particles without affecting interferon cytokines), after which time 0.25 NaOH was added to adjust the pH to 7. In parallel, Vero cells were plated to ˜90% confluence in 96 well format, and the following day incubated with the neutralized media for 24 h prior to infection with Maraba-WT. Interferon secreted from the interrogated cell lines post-Maraba-Δ51 infections would protect the Vero cells from Maraba virus infection, to a degree dependent upon the quantity of interferon produced. After 48 h, survival was quantified using a crystal violet assay (Sigma Aldrich). Briefly, cells were incubated with 1% crystal violet solution, washed, dried, re-suspended in 1% SDS and read at a wavelength of 595 nm.

Interferon responsiveness assay: An indirect “interferon responsive bioassay” was used to estimate the degree to which our cell lines could respond to interferon. PC-3 cells were infected with Maraba-Δ51 (MOI 3) for 18 h to produce interferon, after which time the media was collected and acid neutralized, as described above. The following day, the interferon-containing media was added to the indicated cell lines. Twenty-four hours later, Maraba-WT virus was added at a range of MOIs, and cell viability assays were performed after 48 h of infection. Interferon responsiveness was proportional to the amount of protection conferred by media treatment prior to virus infection.

RT-PCR for XBP1 slicing: Total RNA was extracted from cells using a standard RNeasy spin column kit, as described by the manufacturer (Qiagen). RNA was reverse transcribed to cDNA using Superscript II RT (Invitrogen) following the manufacturer's guidelines. Standard PCR was performed using the following primers: XBP1-F: 5′-cct tgt agt tga gaa cca gg-3′ (SEQ ID NO: 11); XBP1-R; 5′-ggg get tgg tat ata tgt gg-3′ (SEQ ID NO: 12). The PCR products were run out on a 3% agarose gel and visualized with UV imager.

Phase-contrast and fluorescent microscopy: All microscopy was done using a standard dissecting microscope (Nikon SMZ1500). Images were captured using a digital camera (Nikon DXM1200F), and analyzed using computerized software (Nikon ACT software).

Single-step growth curves: U373 cells were seeded into 6-well format at ˜50% confluence, and siRNA transfections were performed the following day. After 72 h, the cells were infected with wild-type Maraba at a multiplicity of infection of 5 pfu/cell for 1 hour. Cells were then washed with PBS and incubated at 37° C. Aliquots (100 μl) were taken at 0, 4, 8, 12, 24, and 48 h time points and titred on Vero cells using a standard plaque assay.

Plaque assays: Vero cells were plated at a density of 5e5 cells per/well of a 6 well dish. The next day, 100 μof serial viral dilutions were prepared and added for 1 hour to Vero cells. After viral adsorption, 2 ml of agarose overlay was added (1:1 1% agarose: 2×DMEM and 20% FCS). Plaques were counted the following day.

Small molecule synthesis: Compounds were synthesized through slight modifications of the methods described in W02008154484. A representative example is given for the synthesis of Compound 2.

Synthesis of Compound 2: A solution of 5.0 g (21.6 mmol) of 5-bromo-2-hydroxy-3-methoxybenzaldehyde, 1.81 mL (1.91 g, 23.8 mmol) of methoxymethyl chloride and 7.53 mL (5.59 g) of diisopropylethylamine (43.28 mmol) of diisopropylethylamine was stirred at ambient temperature in 90 mL of dichloromethane for 3 days. The mixture was concentrated and purified by silica gel chromatography eluting with a gradient of hexanes/ethyl acetate to supply 5.37 g of 5-bromo-3-methoxy-2-(methoxymethoxy)benzaldehyde. A portion of this material (1.0 g, 3.64 mmol) was combined with (3-carbamoylphenyl)boronic acid (0.731 g, 3.64 mmol), potassium phosphate (0.655 g, 6.18 mmol), Pd₂(dba)₃ (33.3 mg, 0.0364 mmol), tricyclohexylphosphine (24.5 mg, 0.0872 mmol), 1,4-dioxane (12.0 mL), and water (6.0 mL) in a microwave vessel and heated in a microwave apparatus for 30 min at 85 deg C. After cooling, the crude reaction mixture was filtered through a pad of Celite, absorbed on to silica gel and purified by silica gel chromatography eluting with 100% ethyl acetate. Fractions showing product were combined and concentrated to give 1.10 g of pure 3′-formyl-5′-methoxy-4′-(methoxymethoxy)-[1,1′-biphenyl]-3-carboxamide. The completion of the synthesis of compound 2 was carried out by dissolving this material in 20 mL tetrahydrofuran and adding 20 mL of 1 N aq. HCl. The mixture was stirred at room temperature under positive nitrogen pressure for 16 h. A yellow precipitate was collected by suction filtration to give, after air drying, the crude product. Trituration using methanol provided 0.59 g of 3′-formyl-4′-hydroxy-5′-methoxy-[1,1′-biphenyl]-3-carboxamide (Compound 2). Analytical data (proton NMR and low resolution electrospray mass spectrometry) was consistent with pure desired product.

Small molecule screening: U373 cells were plated in 6-well format to a confluence of ˜75%. The following day, candidate small molecules were dissolved in DMSO and added directly to the cell culture media at a range of concentrations. After 2 h, tunicamycin (5 μg/mL) was added, and total RNA was collected 4 h later. RNA extraction and RT-PCR for XBP1 splicing were performed as described above.

In vitro cytotoxicity assays with small molecules: Cells were seeded onto 96 well plates to a confluence of ˜50%. The following day, siRNA transfections were performed, or small-molecule IRE1α inhibition was initiated. For the small-molecules, DMSO was used as a vehicle with a [drug]=20-50 μM. Drug treatment occurred for either 4 h (“acute” treatment), or was re-applied at 24 h and left for 48 h total (“chronic” treatment). Following knockdown or chemical inhibition, the cells were infected at log dilutions with the indicated rhabdoviruses. After 48-72 h of infection (depending on the cell line), Resazurin sodium salt was added to a final concentration of 20 μg/ml. After a 6 h incubation the absorbance was read at a wavelength of 573 nm.

Maximum tolerable dose (MTD) and pharmacokinetic (PK) studies in mice: For MTD studies, groups of three CD-1 nude mice (6-8 weeks old) were given a single intraperitoneal (IP) injection of Compound 2 (in log 2 increments, diluted in 10% Tween-80) ranging from −50-1000 mg/Kg. The animals were monitored twice daily for signs of distress, including weight loss, morbidity, and respiratory distress. For PK studies, groups of three CD-1 nude mice (6-8 weeks old) were given a single IP injection of Compound 2 (250 mg/Kg), and blood was taken from the saphenous vein at the indicated timepoints. The blood was centrifuged at 3,000 rpm for 10 min, and plasma collected and frozen (−80 deg C.). Plasma samples were analyzed for Compound 2 using LC-MS. To 10 μL plasma, 20 μL acetonitrile was added, vortexed briefly and centrifuged at 14,000 rpm for 10 min. The clear supernatant was transferred in to a vial for LC-MS analysis. Chromatographic separations were carried out on an Acquity UPLC BEH C18 (2.1×50 mm, 1.7 pm) column using ACQUITY UPLC system. The mobile phase was 0.1% formic acid in water (solvent A) and 0.1% formic acid in acetonitrile (solvent B). A gradient starting at 95% solvent A going to 5% in 4.5 min, holding for 0.5 min, going back to 95% in 0.5 min and equilibrating the column for 1 min was employed. A Waters Xevo QTof MS equipped with an atmospheric pressure ionization source was used for MS analysis. MassLynx 4.1 was used for data analysis. WinNonlin was used to obtain the pharmacokinetic parameters.

Ovarian xenograft model: Human ovarian carcinoma-derived OVCAR-4 cells, adapted for bioluminescent imaging, were injected into 6-8 week old athymic CD-1 nude mice (IP injection, 5×10⁶ cells per mouse). Untreated animals develop measurable abdominal tumours (assessed by IVIS imaging) by 4-7 days, become icteric by 3-4 months and must be euthanized shortly thereafter due to systemic disease as characterized by enlarged cancerous liver and spleen, pale kidneys, and cancerous lymph nodes on the abdominal mesentery. For efficacy experiments, Compound 2 (250 mg/kg; or vehicle (10% Tween-80)) was administered twice daily (IP injections), beginning on day 14 and ending on day 19. Maraba-DM (1×10⁵ pfu per cell) or PBS was injected IV (tail vein) on day 16, 17, 19, 23, 25, 27. Animals were monitored daily for weight loss, morbidity, hind leg paralysis and respiratory distress. Tumour images were captured twice weekly with a Xenogen 200 IVIS system (Caliper LS, USA), and total luminescent flux was analyzed on computerized software (Xenogen).

EMT6 syngeneic breast cancer models: Murine EMT6 breast cancer-derived cells (1×10⁵ per mouse) were adapted for bioluminescent imaging and injected into the fat pad of the right lower breast in 6-8 week old Balb/c mice. Mice developed palpable tumours by 5-7 days, which grew rapidly. At 7 days post-tumour implants, mice were treated with Compound 2 (250 mg/kg, IP) or vehicle, twice daily, for six days. Maraba-DM (1×10⁷ pfu per cell) or PBS was injected IV (tail vein) on day 10, 11, 13, 16, and 17. Animals were monitored daily for piloerection, weight loss, morbidity, hind leg paralysis and respiratory distress. Tumour images were captured twice weekly using a Xenogen 200 IVIS system (Caliper LS, USA), and total luminescent flux was analyzed on computerized software (Xenogen). Mice were euthanized when the total flux exceeded 1×10⁷, which corresponded to a tumour burden of ˜500 mm³ and occurred between 14-17 days in untreated animals.

Statistical analyses: For all statistical analyses except survival curves, one- and two-way ANOVAs were performed followed by a Bonferroni multiple comparison's post-hoc test to derive P values (GraphPad Prism). For survival curves, Mantel-Cox Log rank analysis was used to compare plots (GraphPad Prism).

TABLE 2
Composite list of synthetic lethal hits derived from 3 tumour cell lines
ENTREZ		GENE	COMPOSITE MAD SCORE
GENEID	GENE NAME	SYMBOL	OVCAR8	U373	NCIH226
55016	MEMBRANE-ASSOCIATED RING FINGER (C3HC4) 1	38776		−2.41151	−4.10084
10801	SEPTIN 9	38968	−2.13128	−3.66817	−2.86592
346288	FLJ44060 PROTEIN	38973	−5.54837	−3.0341
51166	AMINOADIPATE AMINOTRANSFERASE	AADAT	−2.49835	−2.95805
22848	AP2 ASSOCIATED KINASE 1	AAK1	−2.77645	−2.01405
14	ANGIO-ASSOCIATED, MIGRATORY CELL PROTEIN	AAMP	−3.11061	−2.60732	−2.15765
15	ARYLALKYLAMINE N-ACETYLTRANSFERASE	AANAT	−1.95955	−2.41831
60496	DKFZP566E2346 PROTEIN	AASDHPPT	−2.16412	−2.38839
23456	ATP-BINDING CASSETTE, SUB-FAMILY B (MDR/TAP), MEMBER 10	ABCB10	−2.45154	−2.10888	−2.11658
6833	ATP-BINDING CASSETTE, SUB-FAMILY C (CFTR/MRP), MEMBER 8	ABCC8	−2.20353	−1.96417
64137	ATP-BINDING CASSETTE, SUB-FAMILY G (WHITE), MEMBER 4	ABCG4	−2.13289	−2.17044
80325	ANKYRIN REPEAT AND BTB (POZ) DOMAIN CONTAINING 1	ABTB1	−2.29777	−2.27845	−1.93118
65057	ADRENOCORTICAL DYSPLASIA HOMOLOG (MOUSE)	ACD	−3.49186	−4.48169
130013	AMINOCARBOXYMUCONATE SEMIALDEHYDE DECARBOXYLASE	ACMSD	−3.479	−3.14302	−2.42407
134526	ACYL-COA THIOESTERASE 12	ACOT12	−1.99657		−3.33543
8309	ACYL-COENZYME A OXIDASE 2, BRANCHED CHAIN	ACOX2	−3.07964	−3.9036	−3.83209
55289	ACYL-COENZYME A OXIDASE-LIKE	ACOXL	−2.18631	−2.45197
10121	ARP1 ACTIN-RELATED PROTEIN 1 HOMOLOG A, CENTRACTIN ALPHA	ACTR1A	−3.02305	−3.39827	−4.92131
	(YEAST)
8747	ADAM METALLOPEPTIDASE DOMAIN 21	ADAM21	−1.93391	−4.53126	−2.64672
80070	ADAM METALLOPEPTIDASE WITH THROMBOSPONDIN TYPE 1 MOTIF, 20	ADAMTS20	−2.21441	−1.90698
23536	ADENOSINE DEAMINASE, TRNA-SPECIFIC 1	ADAT1	−1.93515	−2.22345
107	ADENYLATE CYCLASE 1 (BRAIN)	ADCY1		−2.87709	−2.5232
111	ADENYLATE CYCLASE 5	ADCY5	−2.69979	−2.68721
123	ADIPOSE DIFFERENTIATION-RELATED PROTEIN	ADFP	−5.03679	−2.31512	−2.44941
133	ADRENOMEDULLIN	ADM	−2.40943	−2.44641
84890	CHROMOSOME 10 OPEN READING FRAME 22	ADO	−3.38911	−4.48099
140	ADENOSINE A3 RECEPTOR	ADORA3	−2.5352		−2.76387
173	AFAMIN	AFM	−2.2435	−2.28448
10598	AHA1, ACTIVATOR OF HEAT SHOCK 90 KDA PROTEIN ATPASE	AHSA1	−2.88922	−3.59496	−2.99421
	HOMOLOG 1 (YEAST)
326	AUTOIMMUNE REGULATOR (AUTOIMMUNE POLYENDOCRINOPATHY	AIRE	−2.20195	−3.22496
	CANDIDIASIS ECTODERMAL DYSTROPHY)
8852	A KINASE (PRKA) ANCHOR PROTEIN 4	AKAP4		−2.04143	−2.57289
6718	ALDO-KETO REDUCTASE FAMILY 1, MEMBER D1 (DELTA 4-3-	AKR1D1	−2.07303	−4.60704
	KETOSTEROID-5-BETA-REDUCTASE)
80216	KIAA1527 PROTEIN	ALPK1	−1.95916	−3.47659
151254	AMYOTROPHIC LATERAL SCLEROSIS 2 (JUVENILE) CHROMOSOME	ALS2CR11	−4.75935	−2.53681	−2.40034
	REGION, CANDIDATE 11
258	AMELOBLASTIN, ENAMEL MATRIX PROTEIN	AMBN		−3.16187	−2.15822
348094	ANKYRIN REPEAT AND DEATH DOMAIN CONTAINING 1A	ANKDD1A	−2.00511		−1.87831
81573	ANKYRIN REPEAT DOMAIN 13C	ANKRD13C	−2.89415	−2.23827
84250	ANKYRIN REPEAT DOMAIN 32	ANKRD32		−2.60824	−3.37461
375248	ANKYRIN REPEAT DOMAIN 36	ANKRD36		−2.80648	−2.14125
65124	CHROMOSOME 2 OPEN READING FRAME 26	ANKRD57	−2.19538		−2.57045
307	ANNEXIN A4	ANXA4	−2.47948	−2.7168
164	ADAPTOR-RELATED PROTEIN COMPLEX 1, GAMMA 1 SUBUNIT	AP1G1		−2.98113	−3.03354
160	ADAPTOR-RELATED PROTEIN COMPLEX 2, ALPHA 1 SUBUNIT	AP2A1	−2.0658	−4.29449	−2.4135
161	ADAPTOR-RELATED PROTEIN COMPLEX 2, ALPHA 2 SUBUNIT	AP2A2	−2.29654	−3.52961
11154	ADAPTOR-RELATED PROTEIN COMPLEX 4, SIGMA 1 SUBUNIT	AP4S1	−2.43482	−3.22071	−3.08222
147495	ADENOMATOSIS POLYPOSIS COLI DOWN-REGULATED 1	APCDD1	−3.70917	−3.66013
8539	APOPTOSIS INHIBITOR 5	API5	−2.95141		−2.25814
351	AMYLOID BETA (A4) PRECURSOR PROTEIN (PEPTIDASE NEXIN-II,	APP	−2.21138	−2.30784	−2.14111
	ALZHEIMER DISEASE)
361	AQUAPORIN 4	AQP4		−1.99469	−4.23735
27236	ADP-RIBOSYLATION FACTOR INTERACTING PROTEIN 1 (ARFAPTIN 1)	ARFIP1	−4.11551	−2.80184	−1.9262
392	RHO GTPASE ACTIVATING PROTEIN 1	ARHGAP1	−2.91216	−2.5815
55843	RHO GTPASE ACTIVATING PROTEIN 15	ARHGAP15		−1.97386	−2.25784
84986	RHO GTPASE ACTIVATING PROTEIN 19	ARHGAP19	−2.63195	−2.99612
9138	RHO GUANINE NUCLEOTIDE EXCHANGE FACTOR (GEF) 1	ARHGEF1	−3.08806	−5.5479	−4.87465
340485	N-ACYLSPHINGOSINE AMIDOHYDROLASE 3-LIKE	ASAH3L	−2.97451	−3.77336	−2.38993
79827	ADIPOCYTE-SPECIFIC ADHESION MOLECULE	ASAM	−2.0148		−2.78807
430	ACHAETE-SCUTE COMPLEX-LIKE 2 (DROSOPHILA)	ASCL2	−2.22619	−2.21252	−1.90579
79058	ALVEOLAR SOFT PART SARCOMA CHROMOSOME REGION, CANDIDATE 1	ASPSCR1		−2.00368	−2.08419
22926	ACTIVATING TRANSCRIPTION FACTOR 6	ATF6	−2.79393	−2.93049
491	ATPASE, CA++ TRANSPORTING, PLASMA MEMBRANE 2	ATP2B2	−2.37995	−2.62897
27032	ATPASE, CA++ TRANSPORTING, TYPE 2C, MEMBER 1	ATP2C1	−2.66675	−2.55357	−3.33098
27109	ATP SYNTHASE, H+ TRANSPORTING, MITOCHONDRIAL F0 COMPLEX,	ATP5S	−2.94141		−2.00102
	SUBUNIT S (FACTOR B)
79895	ATPASE, CLASS I, TYPE 8B, MEMBER 4	ATP8B4		−2.32716	−1.96282
11273	ATAXIN 2-LIKE	ATXN2L	−2.16286	−2.90014	−3.3262
9212	AURORA KINASE B	AURKB	−2.13449	−1.86738
10677	ADVILLIN	AVIL	−2.41928	−2.9485
553	ARGININE VASOPRESSIN RECEPTOR 1B	AVPR1B	−1.90893		−2.64188
64651	AXIN1 UP-REGULATED 1	AXUD1	−2.62194	−2.01919
8708	UDP-GAL:BETAGLCNAC BETA 1,3-GALACTOSYLTRANSFERASE,	B3GALT1	−1.87025		−2.13837
	POLYPEPTIDE 1
8707	UDP-GAL:BETAGLCNAC BETA 1,3-GALACTOSYLTRANSFERASE,	B3GALT2	−2.49262	−2.78836
	POLYPEPTIDE 2
11285	XYLOSYLPROTEIN BETA 1,4-GALACTOSYLTRANSFERASE,	B4GALT7	−3.46031	−3.70379
	POLYPEPTIDE 7 (GALACTOSYLTRANSFERASE I)
54971	BTG3 ASSOCIATED NUCLEAR PROTEIN	BANP	−4.91691	−3.03484
55212	BARDET-BIEDL SYNDROME 7	BBS7	−3.48742	−1.98035
64919	B-CELL CLL/LYMPHOMA 11B (ZINC FINGER PROTEIN)	BCL11B		−2.28374	−2.3214
602	B-CELL CLL/LYMPHOMA 3	BCL3	−2.09542		−1.86381
255877	B-CELL CLL/LYMPHOMA 6, MEMBER B (ZINC FINGER PROTEIN)	BCL6B	−2.32078	−2.67158
605	B-CELL CLL/LYMPHOMA 7A	BCL7A		−3.31142	−2.31437
331	BACULOVIRAL IAP REPEAT-CONTAINING 4	BIRC4	−2.8144	−2.34593
652	BONE MORPHOGENETIC PROTEIN 4	BMP4	−2.51609	−2.03764	−2.64151
23246	BLOCK OF PROLIFERATION 1	BOP1	−2.4163	−2.84961
6046	BROMODOMAIN CONTAINING 2	BRD2	−3.70807		−2.19798
56853	BRUNO-LIKE 4, RNA BINDING PROTEIN (DROSOPHILA)	BRUNOL4	−2.38575	−3.7975	−3.28405
138151	BTB (POZ) DOMAIN CONTAINING 14A	BTBD14A	−5.42149	−4.98281	−4.37291
7832	BTG FAMILY, MEMBER 2	BTG2	−2.26701		−2.9354
221060	CHROMOSOME 10 OPEN READING FRAME 111	C10ORF111	−3.52161	−2.1038
55088	CHROMOSOME 10 OPEN READING FRAME 118	C10ORF118		−2.14738	−2.08959
143384	CHROMOSOME 10 OPEN READING FRAME 46	C10ORF46	−2.05153		−2.51401
79741	CHROMOSOME 10 OPEN READING FRAME 68	C10ORF68		−2.00775	−2.09128
79946	CHROMOSOME 10 OPEN READING FRAME 95	C10ORF95	−4.25451	−2.45897
79081	CHROMOSOME 11 OPEN READING FRAME 48	C11ORF48	−6.50729	−3.88765
84067	CHROMOSOME 11 OPEN READING FRAME 56	C11ORF56	−4.29459	−2.6957	−4.52064
745	CHROMOSOME 11 OPEN READING FRAME 9	C11ORF9	−4.1401	−2.58869
283416	CHROMOSOME 12 OPEN READING FRAME 61	C12ORF61	−2.30163	−1.9713	−2.30156
221150	CHROMOSOME 13 OPEN READING FRAME 3	C13ORF3	−3.40649	−4.80828
55668	CHROMOSOME 14 OPEN READING FRAME 118	C14ORF118	−2.21825	−2.63428
64430	CHROMOSOME 14 OPEN READING FRAME 135	C14ORF135	−2.08351	−1.97923
54675	CHROMOSOME 20 OPEN READING FRAME 155	CRLS1	−5.73605	−2.74203
1414	CRYSTALLIN, BETA B1	CRYBB1	−3.42872	−2.10112
1429	CRYSTALLIN, ZETA (QUINONE REDUCTASE)	CRYZ	−2.52933	−3.09796	−2.20554
8531	COLD SHOCK DOMAIN PROTEIN A	CSDA	−2.30656	−5.08058	−2.30271
1437	COLONY STIMULATING FACTOR 2 (GRANULOCYTE-MACROPHAGE)	CSF2		−3.23829	−2.10723
1452	CASEIN KINASE 1, ALPHA 1	CSNK1A1	−2.31878	−2.18873
122011	CASEIN KINASE 1, ALPHA 1-LIKE	CSNK1A1L		−1.85463	−1.90398
1485	CANCER/TESTIS ANTIGEN 1B	CTAG1B	−2.16637		−1.86576
56474	CTP SYNTHASE II	CTPS2	−2.45539	−2.23267	−2.90417
1519	CATHEPSIN O	CTSO	−2.81322		−3.55489
55917	CTTNBP2 N-TERMINAL LIKE	CTTNBP2NL	−2.69729		−2.50301
2919	CHEMOKINE (C—X—C MOTIF) LIGAND 1 (MELANOMA GROWTH	CXCL1	−2.43419	−3.48441
	STIMULATING ACTIVITY, ALPHA)
80319	CXXC FINGER 4	CXXC4	−3.34186	−3.9532
1588	CYTOCHROME P450, FAMILY 19, SUBFAMILY A, POLYPEPTIDE 1	CYP19A1	−2.68268	−2.25115	−2.76115
51302	CYTOCHROME P450, FAMILY 39, SUBFAMILY A, POLYPEPTIDE 1	CYP39A1	−2.56803	−1.99217
284541	CYTOCHROME P450, FAMILY 4, SUBFAMILY A, POLYPEPTIDE 22	CYP4A22		−3.73785	−3.26585
27351	DNA SEGMENT, CHR 15, WAYNE STATE UNIVERSITY 75, EXPRESSED	D15WSU75E	−2.14509		−1.88014
2532	DUFFY BLOOD GROUP, CHEMOKINE RECEPTOR	DARC	−2.71008	−4.47006
440097	DEVELOPING BRAIN HOMEOBOX 2	DBX2	−4.3273	−3.66309
1632	DODECENOYL-COENZYME A DELTA ISOMERASE (3,2 TRANS-ENOYL-	DCI	−2.35369	−1.99944
	COENZYME A ISOMERASE)
9201	DOUBLECORTIN AND CAM KINASE-LIKE 1	DCLK1		−2.5543	−2.68774
64421	DNA CROSS-LINK REPAIR 1C (PSO2 HOMOLOG, S. CEREVISIAE)	DCLRE1C	−1.96141		−2.11474
1638	DOPACHROME TAUTOMERASE (DOPACHROME DELTA-ISOMERASE,	DCT	−2.41936		−2.71413
	TYROSINE-RELATED PROTEIN 2)
55208	DCN1, DEFECTIVE IN CULLIN NEDDYLATION 1, DOMAIN CONTAINING 2	DCUN1D2	−1.93227	−1.90673
	(S. CEREVISIAE)
80821	DDHD DOMAIN CONTAINING 1	DDHD1		−2.0493	−2.87835
84301	DDI1, DNA-DAMAGE INDUCIBLE 1, HOMOLOG 2 (S. CEREVISIAE)	DDI2	−4.31245	−2.95685	−3.83909
55510	DEAD (ASP-GLU-ALA-ASP) BOX POLYPEPTIDE 43	DDX43		−2.24216	−3.00093
10522	DEFORMED EPIDERMAL AUTOREGULATORY FACTOR 1 (DROSOPHILA)	DEAF1	−2.17474	−1.91778
1668	DEFENSIN, ALPHA 1	DEFA3	−4.79547	−2.23812
414325	DEFENSIN, BETA 103B	DEFB103B	−3.21911	−1.92634
140850	DEFENSIN, BETA 127	DEFB127	−2.55821	−2.07617
8560	DEGENERATIVE SPERMATOCYTE HOMOLOG 1, LIPID DESATURASE	DEGS1	−2.11252	−3.53413
	(DROSOPHILA)
79139	DER1-LIKE DOMAIN FAMILY, MEMBER 1	DERL1	−2.3016	−1.87201
51009	DER1-LIKE DOMAIN FAMILY, MEMBER 2	DERL2	−3.0246	−1.95458
1676	DNA FRAGMENTATION FACTOR, 45 KDA, ALPHA POLYPEPTIDE	DFFA	−2.81706	−4.22012	−3.21915
1677	DNA FRAGMENTATION FACTOR, 40 KDA, BETA POLYPEPTIDE	DFFB	−2.12535	−3.3819
	(CASPASE-ACTIVATED DNASE)
85359	DIGEORGE SYNDROME CRITICAL REGION GENE 6-LIKE	DGCR6L	−3.38017	−2.23632
8526	DIACYLGLYCEROL KINASE, EPSILON 64 KDA	DGKE	−3.20331	−2.93058
27294	DIHYDRODIOL DEHYDROGENASE (DIMERIC)	DHDH	−2.3481	−2.28125
23181	DIP2 DISCO-INTERACTING PROTEIN 2 HOMOLOG A (DROSOPHILA)	DIP2A	−2.18537	−2.41581	−2.37741
27123	DICKKOPF HOMOLOG 2 (XENOPUS LAEVIS)	DKK2	−3.65488		−2.91351
9231	DISCS, LARGE HOMOLOG 5 (DROSOPHILA)	DLG5	−2.197	−2.7603
1750	DISTAL-LESS HOMEOBOX 6	DLX6		−1.86594	−2.43545
55567	DYNEIN, AXONEMAL, HEAVY POLYPEPTIDE 3	DNAH3	−2.27884		−2.05462
1769	DYNEIN, AXONEMAL, HEAVY POLYPEPTIDE 8	DNAH8	−2.05019		−3.08662
85479	DNAJ (HSP40) HOMOLOG, SUBFAMILY C, MEMBER 5 BETA	DNAJC5B	−3.74895	−2.7117
144132	DYNEIN HEAVY CHAIN DOMAIN 1	DNHD1	−1.86308		−1.85855
57572	DEDICATOR OF CYTOKINESIS 6	DOCK6	−2.50006		−2.61583
1797	DOM-3 HOMOLOG Z (C. ELEGANS)	DOM3Z	−2.77576	−1.97913
84444	DOTI-LIKE, HISTONE H3 METHYLTRANSFERASE (S. CEREVISIAE)	DOT1L		−3.22467	−2.39116
1801	DPH1 HOMOLOG (S. CEREVISIAE)	DPH1	−2.78644	−3.04536	−2.96731
54344	DOLICHYL-PHOSPHATE MANNOSYLTRANSFERASE POLYPEPTIDE 3	DPM3		−2.62437	−2.25698
340168	DEVELOPMENTAL PLURIPOTENCY ASSOCIATED 5	DPPA5	−4.16294	−2.28648	−2.10864
1807	DIHYDROPYRIMIDINASE	DPYS	−1.99033	−4.15073
1812	DOPAMINE RECEPTOR D1	DRD1	−2.6749	−1.91894	−2.65582
1814	DOPAMINE RECEPTOR D3	DRD3	−4.81514	−3.38421
1826	DOWN SYNDROME CELL ADHESION MOLECULE	DSCAM	−1.8794	−2.99691
1828	DESMOGLEIN 1	DSG1	−1.86862	−3.18021
1832	DESMOPLAKIN	DSP	−4.3071		−2.18346
80824	DUAL SPECIFICITY PHOSPHATASE 16	DUSP16	−2.10263	−2.55858
63904	DUAL SPECIFICITY PHOSPHATASE 21	DUSP21		−3.15806	−2.5002
84332	HYPOTHETICAL PROTEIN MGC16186	DYDC2		−2.38669	−2.35023
1783	DYNEIN, CYTOPLASMIC 1, LIGHT INTERMEDIATE CHAIN 2	DYNC1LI2	−4.31596	−2.06078
1859	DUAL-SPECIFICITY TYROSINE-(Y)-PHOSPHORYLATION REGULATED	DYRK1A	−2.69878	−3.14822	−1.97728
	KINASE 1A
1877	E4F TRANSCRIPTION FACTOR 1	E4F1	−2.17668		−2.6409
1889	ENDOTHELIN CONVERTING ENZYME 1	ECE1	−2.5034	−2.16589
79746	ENOYL COENZYME A HYDRATASE DOMAIN CONTAINING 3	ECHDC3	−3.73688	−4.15934
112399	HYPOTHETICAL PROTEIN FLJ21620	EGLN3	−2.76447	−4.40225
126272	EID-2-LIKE INHIBITOR OF DIFFERENTIATION-3	EID2B	−3.34573	−2.33512	−2.44539
440275	EUKARYOTIC TRANSLATION INITIATION FACTOR 2 ALPHA KINASE 4	EIF2AK4		−4.47732	−3.08146
7458	WILLIAMS-BEUREN SYNDROME CHROMOSOME REGION 1	EIF4H	−3.13161	−4.5717	−2.33113
23436	ELASTASE 3A, PANCREATIC	ELA3B	−2.87901	−2.22243
114794	KIAA1904 PROTEIN	ELFN2	−3.93848	−3.94671
10436	EMG1 NUCLEOLAR PROTEIN HOMOLOG (S. CEREVISIAE)	EMG1	−4.78692	−4.19447
2009	ECHINODERM MICROTUBULE ASSOCIATED PROTEIN LIKE 1	EML1	−4.52565		−2.91035
27436	ECHINODERM MICROTUBULE ASSOCIATED PROTEIN LIKE 4	EML4		−3.38095	−1.99126
9941	ENDONUCLEASE G-LIKE 1	ENDOGL1	−2.34432	−1.91204	−2.4438
55068	PROLIFERATION-INDUCING PROTEIN 38	ENOX1	−3.15056	−5.20001	−2.74843
339221	ECTONUCLEOTIDE PYROPHOSPHATASE/PHOSPHODIESTERASE 7	ENPP7		−3.43628	−1.85947
957	ECTONUCLEOSIDE TRIPHOSPHATE DIPHOSPHOHYDROLASE 5	ENTPD5	−5.07624	−3.65826
56943	ENHANCER OF YELLOW 2 HOMOLOG (DROSOPHILA)	ENY2	−1.94151		−2.25661
2034	ENDOTHELIAL PAS DOMAIN PROTEIN 1	EPAS1	−2.32691	−3.32114	−2.88588
54566	ERYTHROCYTE MEMBRANE PROTEIN BAND 4.1 LIKE 4B	EPB41L4B	−2.20157		−3.22323
2044	EPH RECEPTOR A5	EPHA5	−4.13072	−4.98528	−5.66157
55040	EPSIN 3	EPN3	−2.50122		−2.18479
2057	ERYTHROPOIETIN RECEPTOR	EPOR	−1.96407	−2.89036	−2.10795
2067	EXCISION REPAIR CROSS-COMPLEMENTING RODENT REPAIR	ERCC1	−3.30242	−3.35171
	DEFICIENCY, COMPLEMENTATION GROUP 1 (INCLUDES OVERLAPPING
	ANTISENSE SEQUENCE)
2068	EXCISION REPAIR CROSS-COMPLEMENTING RODENT REPAIR	ERCC2		−2.87085	−3.83979
	DEFICIENCY, COMPLEMENTATION GROUP 2 (XERODERMA
	PIGMENTOSUM D)
2081	ENDOPLASMIC RETICULUM TO NUCLEUS SIGNALLING 1	ERN1	−3.26923	−2.48179
345062	EPIDERMIS-SPECIFIC SERINE PROTEASE-LIKE PROTEIN	ESSPL	−2.38312	−2.17637
2128	EVE, EVEN-SKIPPED HOMEOBOX HOMOLOG 1 (DROSOPHILA)	EVX1		−2.94372	−3.14417
2153	COAGULATION FACTOR V (PROACCELERIN, LABILE FACTOR)	F5		−3.25947	−3.02546
2172	FATTY ACID BINDING PROTEIN 6, ILEAL (GASTROTROPIN)	FABP6	−4.08486	−3.59984	−1.94519
3992	FATTY ACID DESATURASE 1	FADS1	−4.03858	−3.83463
151313	HYPOTHETICAL PROTEIN DKFZP434N062	FAHD2B	−3.11705	−3.22452	−2.59929
9747	KIAA0738 GENE PRODUCT	FAM115A	−3.84046	−2.7195	−2.45971
81558	C/EBP-INDUCED PROTEIN	FAM117A		−4.43236	−2.04035
54855	FAMILY WITH SEQUENCE SIMILARITY 46, MEMBER C	FAM46C	−2.81984		−2.89087
442444	SIMILAR TO HYPOTHETICAL PROTEIN FLJ35782	FAM47C	−2.36742	−2.31443	−1.86826
113115	FAMILY WITH SEQUENCE SIMILARITY 54, MEMBER A	FAM54A	−2.3527	−1.94078
91775	FAMILY WITH SEQUENCE SIMILARITY 55, MEMBER C	FAM55C		−1.90321	−1.90662
149297	FAMILY WITH SEQUENCE SIMILARITY 78, MEMBER B	FAM78B		−5.19784	−2.27141
2177	FANCONI ANEMIA, COMPLEMENTATION GROUP D2	FANCD2	−2.59596		−2.19731
2191	FIBROBLAST ACTIVATION PROTEIN, ALPHA	FAP	−1.90529	−2.09759
10160	FERM, RHOGEF (ARHGEF) AND PLECKSTRIN DOMAIN PROTEIN 1	FARP1	−2.25154	−2.07884
	(CHONDROCYTE-DERIVED)
2196	FAT TUMOR SUPPRESSOR HOMOLOG 2 (DROSOPHILA)	FAT2	−1.92073	−2.64341
54751	FILAMIN BINDING LIM PROTEIN 1	FBLIM1	−2.35839		−2.6936
129804	HYPOTHETICAL PROTEIN FLJ37440	FBLN7		−2.02407	−2.19875
22992	F-BOX AND LEUCINE-RICH REPEAT PROTEIN 11	FBXL11		−2.33502	−3.60892
54620	F-BOX AND LEUCINE-RICH REPEAT PROTEIN 19	FBXL19	−2.29513		−2.27125
126433	F-BOX PROTEIN 27	FBXO27		−2.11258	−2.49207
26259	F-BOX AND WD-40 DOMAIN PROTEIN 8	FBXW8	−2.80401		−2.13207
83953	FC RECEPTOR, IGA, IGM, HIGH AFFINITY	FCAMR	−4.09839	−1.9096	−2.07656
9103	FC FRAGMENT OF IGG, LOW AFFINITY IIC, RECEPTOR FOR (CD32)	FCGR2C		−2.48481	−2.28736
2865	FREE FATTY ACID RECEPTOR 3	FFAR3	−3.5176		−2.17937
9457	FOUR AND A HALF LIM DOMAINS 5	FHL5	−3.72762	−2.80553
2307	FORKHEAD-LIKE 18 (DROSOPHILA)	FKHL18	−2.99411	−3.40374
388939	SIMILAR TO CDNA SEQUENCE BC027072	FLJ34931	−3.78588	−2.21798
222183	HYPOTHETICAL PROTEIN FLJ37078	FLJ37078	−2.99604		−2.8664
643853	SIMILAR TO F40B5.2B	FLJ45032	−2.2066	−2.91152
440107	FLJ46688 PROTEIN	FLJ46688	−2.94624	−2.03724
23769	FIBRONECTIN LEUCINE RICH TRANSMEMBRANE PROTEIN 1	FLRT1	−3.78841	−2.82884
2324	FMS-RELATED TYROSINE KINASE 4	FLT4	−1.93582	−2.52564
2348	FOLATE RECEPTOR 1 (ADULT)	FOLR1	−1.87156	−4.52012
2350	FOLATE RECEPTOR 2 (FETAL)	FOLR2	−2.22889	−3.43517
442425	SIMILAR TO FOXB2 PROTEIN	FOXB2	−3.4805	−2.17358
22887	FORKHEAD BOX J3	FOXJ3		−3.34354	−1.97325
93986	TRINUCLEOTIDE REPEAT CONTAINING 10	FOXP2	−3.44945	−2.52173	−2.81991
2487	FRIZZLED-RELATED PROTEIN	FRZB		−2.18499	−1.93131
2492	FOLLICLE STIMULATING HORMONE RECEPTOR	FSHR	−3.11704	−2.88641	−3.92828
10468	FOLLISTATIN	FST	−4.64815	−2.97286	−4.3585
2528	FUCOSYLTRANSFERASE 6 (ALPHA (1,3) FUCOSYLTRANSFERASE)	FUT6		−2.28549	−2.97626
2533	FYN BINDING PROTEIN (FYB-120/130)	FYB	−1.97259		−2.54165
2535	FRIZZLED HOMOLOG 2 (DROSOPHILA)	FZD2	−1.89392		−2.92839
139716	GRB2-ASSOCIATED BINDING PROTEIN 3	GAB3		−4.11116	−2.31614
2562	GAMMA-AMINOBUTYRIC ACID (GABA) A RECEPTOR, BETA 3	GABRB3	−2.5385	−2.00785
130589	GALACTOSE MUTAROTASE (ALDOSE 1-EPIMERASE)	GALM		−2.21926	−2.53267
2588	GALACTOSAMINE (N-ACETYL)-6-SULFATE SULFATASE (MORQUIO	GALNS	−2.64464		−1.87302
	SYNDROME, MUCOPOLYSACCHARIDOSIS TYPE IVA)
51809	UDP-N-ACETYL-ALPHA-D-GALACTOSAMINE:POLYPEPTIDE N-	GALNT7		−2.28686	−2.24023
	ACETYLGALACTOSAMINYLTRANSFERASE 7 (GALNAC-T7)
117248	UDP-N-ACETYL-ALPHA-D-GALACTOSAMINE:POLYPEPTIDE N-	GALNTL2	−3.01111	−2.98456	−3.25224
	ACETYLGALACTOSAMINYLTRANSFERASE-LIKE 2
8522	GROWTH ARREST-SPECIFIC 7	GAS7	−2.53612	−2.54686	−3.13161
115361	GUANYLATE BINDING PROTEIN 4	GBP4	−2.68608	−2.62916
9247	GLIAL CELLS MISSING HOMOLOG 2 (DROSOPHILA)	GCM2	−4.05623	−1.99879
151449	GROWTH DIFFERENTIATION FACTOR 7	GDF7	−3.36855	−1.93765	−2.16948
2664	GDP DISSOCIATION INHIBITOR 1	GDI1	−2.87815	−3.44471	−3.453
199720	GAMETOGENETIN	GGN	−3.75431	−2.10603	−3.11222
2693	GROWTH HORMONE SECRETAGOGUE RECEPTOR	GHSR	−1.87572	−2.68113
54826	HYPOTHETICAL PROTEIN FLJ20125	GIN1	−3.47563	−3.9474
169792	GLIS FAMILY ZINC FINGER 3	GLIS3	−2.8858	−2.03553	−1.9006
9340	GLUCAGON-LIKE PEPTIDE 2 RECEPTOR	GLP2R	−2.76923	−3.31118	−2.45118
2752	GLUTAMATE-AMMONIA LIGASE (GLUTAMINE SYNTHETASE)	GLUL	−1.9631		−1.86752
2769	GUANINE NUCLEOTIDE BINDING PROTEIN (G PROTEIN), ALPHA 15 (GQ	GNA15	−2.40069	−2.58242
	CLASS)
2778	GNAS COMPLEX LOCUS	GNAS	−3.95239	−3.13278	−3.19362
2781	GUANINE NUCLEOTIDE BINDING PROTEIN (G PROTEIN), ALPHA Z	GNAZ	−4.24008	−2.58879	−3.72885
	POLYPEPTIDE
2787	GUANINE NUCLEOTIDE BINDING PROTEIN (G PROTEIN), GAMMA 5	GNG5		−3.99978	−3.03387
2794	GUANINE NUCLEOTIDE BINDING PROTEIN-LIKE 1	GNL1	−3.26378		−2.57438
55638	HYPOTHETICAL PROTEIN FLJ20366	GOLSYN	−1.91757		−2.45471
54856	GON-4-LIKE (C. ELEGANS)	GON4L	−2.56712	−3.21962
64689	GOLGI REASSEMBLY STACKING PROTEIN 1, 65 KDA	GORASP1	−2.65971	−3.85635
8733	GLYCOSYLPHOSPHATIDYLINOSITOL ANCHOR ATTACHMENT PROTEIN 1	GPAA1	−4.74693	−2.20232
	HOMOLOG (YEAST)
2239	GLYPICAN 4	GPC4		−3.67898	−2.66778
56927	G PROTEIN-COUPLED RECEPTOR 108	GPR108	−3.65187	−1.91374
266977	HYPOTHETICAL PROTEIN FLJ22684	GPR110	−3.10349	−3.28571
283383	G PROTEIN-COUPLED RECEPTOR 133	GPR133	−2.62395		−1.87835
124274	G PROTEIN-COUPLED RECEPTOR 139	GPR139	−2.32763		−3.7658
353345	G PROTEIN-COUPLED RECEPTOR 141	GPR141	−3.53403	−1.94583
4935	G PROTEIN-COUPLED RECEPTOR 143	GPR143		−1.98133	−2.81368
57512	G PROTEIN-COUPLED RECEPTOR 158	GPR158	−4.59117	−2.00912
79581	G PROTEIN-COUPLED RECEPTOR 172A	GPR172A	−2.67151		−1.9873
2866	G PROTEIN-COUPLED RECEPTOR 42	GPR42	−3.58878	−2.52295	−2.31642
10149	G PROTEIN-COUPLED RECEPTOR 64	GPR64	−2.88228	−2.33642
8111	G PROTEIN-COUPLED RECEPTOR 68	GPR68	−4.6542		−2.79728
114928	G PROTEIN-COUPLED RECEPTOR ASSOCIATED SORTING PROTEIN 2	GPRASP2	−2.59236		−2.06134
26086	G-PROTEIN SIGNALLING MODULATOR 1 (AGS3-LIKE, C. ELEGANS)	GPSM1	−2.96143	−2.5368
23708	G1 TO S PHASE TRANSITION 2	GSPT2	−2.08666	−2.49344
79807	HYPOTHETICAL PROTEIN FLJ13273	GSTCD	−1.94122	−1.85446
9328	GENERAL TRANSCRIPTION FACTOR IIIC, POLYPEPTIDE 5, 63 KDA	GTF3C5	−3.29436	−2.28714	−2.33891
474382	H2A HISTONE FAMILY, MEMBER B3	H2AFB3	−3.11566		−2.0306
3066	HISTONE DEACETYLASE 2	HDAC2	−1.85138	−3.34838	−2.95007
8841	HISTONE DEACETYLASE 3	HDAC3	−2.55243		−2.05805
3067	HISTIDINE DECARBOXYLASE	HDC	−2.17027		−2.3297
25831	HECT DOMAIN CONTAINING 1	HECTD1	−2.9087		−1.91184
57520	HECT, C2 AND WW DOMAIN CONTAINING E3 UBIQUITIN PROTEIN	HECW2	−4.96002	−2.65557
	LIGASE 2
220296	HEPATOCYTE CELL ADHESION MOLECULE	HEPN1	−2.35488	−2.28112
64399	HEDGEHOG INTERACTING PROTEIN	HHIP	−3.09954	−2.78448
3090	HYPERMETHYLATED IN CANCER 1	HIC1	−5.18038	−4.25796	−2.45575
192286	HIG1 DOMAIN FAMILY, MEMBER 2A	HIGD2A	−2.01104	−3.8801
8342	HISTONE 1, H2BM	HIST1H2BM	−3.01447		−2.34978
8352	HISTONE 1, H3A	HIST1H3J	−2.26906	−2.2589	−2.45454
3118	MAJOR HISTOCOMPATIBILITY COMPLEX, CLASS II, DQ ALPHA 1	HLA-	−2.70344	−1.96681
		DQA2
3127	MAJOR HISTOCOMPATIBILITY COMPLEX, CLASS II, DR BETA 1	HLA-	−1.96407	−3.55599
		DRB5
3145	HYDROXYMETHYLBILANE SYNTHASE	HMBS	−2.61436	−2.74885	−1.98454
10473	HIGH MOBILITY GROUP NUCLEOSOMAL BINDING DOMAIN 4	HMGN4	−1.85907	−2.09832
10949	HETEROGENEOUS NUCLEAR RIBONUCLEOPROTEIN A0	HNRNPA0		−2.18951	−2.89115
9455	HOMER HOMOLOG 2 (DROSOPHILA)	HOMER2	−3.9976		−2.6051
3206	HOMEOBOX A10	HOXA10	−1.86466	−3.79496	−3.24011
3219	HOMEOBOX B9	HOXB9		−2.23595	−2.86036
3238	HOMEOBOX D12	HOXD12	−3.21087	−2.52466
3248	HYDROXYPROSTAGLANDIN DEHYDROGENASE 15-(NAD)	HPGD	−1.86577		−2.55677
54979	HRAS-LIKE SUPPRESSOR 2	HRASLS2	−3.15182	−1.89794
117245	HRAS-LIKE SUPPRESSOR FAMILY, MEMBER 5	HRASLS5	−3.33291	−3.60836
3273	HISTIDINE-RICH GLYCOPROTEIN	HRG	−3.18723	−3.4721	−2.06195
64342	HS1-BINDING PROTEIN 3	HS1BP3	−2.96461	−3.11986	−2.4583
90161	HEPARAN SULFATE 6-O-SULFOTRANSFERASE 2	HS6ST2		−2.77556	−2.84445
345275	HYDROXYSTEROID (17-BETA) DEHYDROGENASE 13	HSD17B13	−2.00983	−2.12285
3294	HYDROXYSTEROID (17-BETA) DEHYDROGENASE 2	HSD17B2	−4.49553	−3.43069
8630	HYDROXYSTEROID (17-BETA) DEHYDROGENASE 6	HSD17B6	−3.46952	−2.23407	−3.31334
3356	5-HYDROXYTRYPTAMINE (SEROTONIN) RECEPTOR 2A	HTR2A	−2.29234		−3.52702
23463	ISOPRENYLCYSTEINE CARBOXYL METHYLTRANSFERASE	ICMT	−3.03314	−2.30793
51278	IMMEDIATE EARLY RESPONSE 5	IER5	−1.91326	−2.78467
439996	INTERFERON-INDUCED PROTEIN WITH TETRATRICOPEPTIDE REPEATS	IFIT1L	−2.40962		−2.43055
	1-LIKE
3446	INTERFERON, ALPHA 10	IFNA10	−5.85688	−3.18894
3456	INTERFERON, BETA 1, FIBROBLAST	IFNB1	−2.97838	−2.04472
26160	INTRAFLAGELLAR TRANSPORT 172 HOMOLOG (CHLAMYDOMONAS)	IFT172	−2.03712	−2.47326
3488	INSULIN-LIKE GROWTH FACTOR BINDING PROTEIN 5	IGFBP5		−2.5889	−2.46803
3489	INSULIN-LIKE GROWTH FACTOR BINDING PROTEIN 6	IGFBP6		−1.98094	−2.28982
9641	INHIBITOR OF KAPPA LIGHT POLYPEPTIDE GENE ENHANCER IN B-	IKBKE	−2.71769	−4.20239
	CELLS, KINASE EPSILON
64375	ZINC FINGER PROTEIN, SUBFAMILY 1A, 4 (EOS)	IKZF4	−4.50915	−3.62475
53342	INTERLEUKIN 17D	IL17D	−2.57159	−2.31667
53832	INTERLEUKIN 20 RECEPTOR, ALPHA	IL20RA	−2.19032	−2.76022
9235	INTERLEUKIN 32	IL32	−2.10552		−2.01423
3608	INTERLEUKIN ENHANCER BINDING FACTOR 2, 45 KDA	ILF2	−3.12856	−2.40931
55272	IMP3, U3 SMALL NUCLEOLAR RIBONUCLEOPROTEIN, HOMOLOG	IMP3		−1.9611	−2.08666
	(YEAST)
440068	INHIBITORY CASPASE RECRUITMENT DOMAIN (CARD) PROTEIN	INCA	−3.61419	−2.14377
10022	INSULIN-LIKE 5	INSL5	−1.85932		−2.36095
3642	INSULINOMA-ASSOCIATED 1	INSM1	−4.57994	−4.7159	−2.96116
26512	DKFZP434B105 PROTEIN	INTS6	−2.46922	−3.1278
79711	IMPORTIN 4	IPO4	−2.19665	−2.88652	−3.89452
3656	INTERLEUKIN-1 RECEPTOR-ASSOCIATED KINASE 2	IRAK2	−5.14744	−2.90272	−4.4479
10379	INTERFERON-STIMULATED TRANSCRIPTION FACTOR 3, GAMMA 48 KDA	IRF9	−3.29863	−5.05236	−2.76033
8471	INSULIN RECEPTOR SUBSTRATE 4	IRS4	−4.12096	−3.11419
57611	IMMUNOGLOBULIN SUPERFAMILY CONTAINING LEUCINE-RICH REPEAT 2	ISLR2		−2.67151	−2.3868
8516	INTEGRIN, ALPHA 8	ITGA8	−4.66407	−2.77516
3698	INTER-ALPHA (GLOBULIN) INHIBITOR H2	ITIH2	−1.92876	−3.06384
3712	ISOVALERYL COENZYME A DEHYDROGENASE	IVD		−1.97496	−2.22619
23081	JUMONJI DOMAIN CONTAINING 2C	JMJD2C	−3.5406	−2.84532	−3.65305
56704	JUNCTOPHILIN 1	JPH1	−5.13892	−3.00751	−4.09157
10899	JUMPING TRANSLOCATION BREAKPOINT	JTB		−1.92897	−2.2368
27133	POTASSIUM VOLTAGE-GATED CHANNEL, SUBFAMILY H (EAG-	KCNH5	−2.18888		−2.30075
	RELATED), MEMBER 5
9798	KIAA0174	KIAA0174	−6.04739	−2.65805
9895	KIAA0329	KIAA0329	−2.72568	−3.29382
23334	KIAA0467	KIAA0467	−2.73811		−1.89679
9858	KIAA0649	KIAA0649	−2.87843		−2.07629
57521	RAPTOR	KIAA1303	−2.31287	−2.90653	−2.4201
57650	KIAA1524	KIAA1524	−3.72788	−2.34109
80817	KIAA1712	KIAA1712		−3.59888	−2.5921
85449	KIAA1755 PROTEIN	KIAA1755	−3.70085	−4.56373	−3.39759
90231	KIAA2013	KIAA2013	−2.09224	−2.6924
57576	KINESIN FAMILY MEMBER 17	KIF17		−2.57894	−2.97311
124602	KINESIN FAMILY MEMBER 19	KIF19	−2.11698	−2.1023	−3.33649
9493	KINESIN FAMILY MEMBER 23	KIF23	−2.05	−1.96415
26153	KINESIN FAMILY MEMBER 26A	KIF26A	−2.31143		−2.35145
11278	KRUPPEL-LIKE FACTOR 12	KLF12	−1.862	−2.25234
23588	KELCH DOMAIN CONTAINING 2	KLHDC2	−3.17501	−3.05664
56062	KELCH (DROSOPHILA)-LIKE 4	KLHL4	−2.93844	−2.56313
9622	KALLIKREIN 4 (PROSTASE, ENAMEL MATRIX, PROSTATE)	KLK4		−3.51019	−2.13427
353323	KERATIN ASSOCIATED PROTEIN 12-2	KRTAP12-2		−4.07607	−4.27747
337972	KERATIN ASSOCIATED PROTEIN 19-5	KRTAP19-5	−2.10694	−1.86912	−1.8969
85287	KERATIN ASSOCIATED PROTEIN 4-7	KRTAP4-7	−3.1366	−3.19935
440023	KERATIN ASSOCIATED PROTEIN 5-6	KRTAP5-6		−2.64506	−2.60634
388533	KIPV467	KRTDAP	−3.84172	−3.12836
56983	CHROMOSOME 3 OPEN READING FRAME 9	KTELC1	−2.64341	−3.73793
84456	L(3)MBT-LIKE 3 (DROSOPHILA)	L3MBTL3		−2.78776	−2.27559
3916	LYSOSOMAL-ASSOCIATED MEMBRANE PROTEIN 1	LAMP1		−2.02168	−2.60901
143903	LAYILIN	LAYN		−2.46796	−2.73265
3930	LAMIN B RECEPTOR	LBR	−4.19709	−1.91704
85474	LADYBIRD HOMEOBOX HOMOLOG 2 (DROSOPHILA)	LBX2		−4.38027	−2.79667
353139	LATE CORNIFIED ENVELOPE 2A	LCE2A	−2.31879	−2.94629
84458	LIGAND-DEPENDENT COREPRESSOR	LCOR		−3.16053	−2.74306
11061	LEUKOCYTE CELL DERIVED CHEMOTAXIN 1	LECT1	−2.71543	−3.92097
3965	LECTIN, GALACTOSIDE-BINDING, SOLUBLE, 9 (GALECTIN 9)	LGALS9	−2.1954	−1.90199
10186	LIPOMA HMGIC FUSION PARTNER	LHFP	−2.00938	−2.83484
375612	LIPOMA HMGIC FUSION PARTNER-LIKE 3	LHFPL3	−1.90933	−2.88839
375323	LIPOMA HMGIC FUSION PARTNER-LIKE PROTEIN 4	LHFPL4	−1.96573		−2.02048
3985	LIM DOMAIN KINASE 2	LIMK2		−2.40384	−2.33116
96626	LIM AND SENESCENT CELL ANTIGEN-LIKE DOMAINS 3	LIMS3	−3.87705	−2.19621	−2.81259
64130	LIN-7 HOMOLOG B (C. ELEGANS)	LIN7B	−4.05261		−3.26445
158038	LEUCINE RICH REPEAT NEURONAL 6C	LINGO2	−2.22343	−1.8526	−2.0549
84823	LAMIN B2	LMNB2	−3.57509	−2.2521
8543	LIM DOMAIN ONLY 4	LMO4	−2.89404	−2.082	−2.42817
348801	HYPOTHETICAL PROTEIN LOC348801	LNP1		−2.25674	−2.73949
126075	HYPOTHETICAL PROTEIN LOC126075	LOC126075	−2.45589		−2.93435
153364	SIMILAR TO METALLO-BETA-LACTAMASE SUPERFAMILY PROTEIN	LOC153364	−1.95863	−4.24838	−3.71792
161247	SIMILAR TO CG10671-LIKE	LOC161247	−3.371	−2.40455
162993	HYPOTHETICAL PROTEIN LOC162993	LOC162993	−2.16393	−3.31765	−1.97896
201725	HYPOTHETICAL PROTEIN LOC201725	LOC201725	−5.68546		−3.94516
202459	SIMILAR TO RIKEN CDNA 2310008M10	LOC202459		−1.94148	−3.3487
26010	DNA POLYMERASE-TRANSACTIVATED PROTEIN 6	LOC26010	−1.96021	−2.25198	−2.23318
283677	HYPOTHETICAL LOC283677	LOC283677		−2.64581	−4.19504
338809	HYPOTHETICAL PROTEIN LOC338809	LOC338809	−3.81319	−1.94045
390243	SIMILAR TO FOLATE RECEPTOR 4 (DELTA) ISOFORM 1	LOC390243	−2.36576	−2.36707	−1.89087
399818	SIMILAR TO CG9643-PA	LOC399818	−1.99734	−2.05135
400506	SIMILAR TO TSG118.1	LOC400506	−5.33979		−2.27212
440093	SIMILAR TO H3 HISTONE, FAMILY 3B	LOC440093	−2.02969	−1.88758	−3.3132
441294	SIMILAR TO CTAGE6	LOC441294	−2.26952		−2.2475
51057	HYPOTHETICAL PROTEIN LOC51057	LOC51057	−1.8843		−2.3085
63920	TRANSPOSON-DERIVED BUSTER3 TRANSPOSASE-LIKE	LOC63920	−3.94367	−2.89712
643905	SIMILAR TO PROTOCADHERIN 15B	LOC643905	−1.93464	−2.62212
653192	SIMILAR TO TRIPARTITE MOTIF PROTEIN 17	LOC653192	−1.98189		−3.01251
653319	SIMILAR TO HYPOTHETICAL PROTEIN LOC283849	LOC653319	−2.95378		−2.33583
90835	HYPOTHETICAL PROTEIN LOC90835	LOC90835		−2.72483	−3.6019
84171	LYSYL OXIDASE-LIKE 4	LOXL4		−2.73286	−2.93429
9663	LIPIN 2	LPIN2		−2.54186	−3.12008
79782	LEUCINE RICH REPEAT CONTAINING 31	LRRC31	−2.86285	−1.98006	−2.17554
64101	LEUCINE RICH REPEAT CONTAINING 4	LRRC4	−2.62687	−2.28397
94030	LEUCINE RICH REPEAT CONTAINING 4B	LRRC4B		−3.66397	−3.0838
220074	LEUCINE RICH REPEAT CONTAINING 51	LRRC51	−2.13492	−2.49709
9209	LEUCINE RICH REPEAT (IN FLII) INTERACTING PROTEIN 2	LRRFIP2	−2.61026	−2.36935
338821	ORGANIC ANION TRANSPORTER LST-3B	LST-		−2.46569	−1.88141
		3TM12
51213	LEUCINE ZIPPER PROTEIN 4	LUZP4	−2.34327	−3.27072
27076	LY6/PLAUR DOMAIN CONTAINING 3	LYPD3	−3.6252	−2.24353	−2.00614
130574	HYPOTHETICAL PROTEIN MGC52057	LYPD6	−3.48063	−4.26156
84445	LEUCINE ZIPPER, PUTATIVE TUMOR SUPPRESSOR 2	LZTS2	−3.7582	−2.00544
4081	MAB-21-LIKE 1 (C. ELEGANS)	MAB21L1	−4.44621	−3.48347	−2.45736
84944	MAELSTROM HOMOLOG (DROSOPHILA)	MAEL	−3.34187		−2.07536
9935	V-MAF MUSCULOAPONEUROTIC FIBROSARCOMA ONCOGENE	MAFB		−2.01537	−1.96292
	HOMOLOG B (AVIAN)
4113	MELANOMA ANTIGEN FAMILY B, 2	MAGEB2	−1.9079		−2.59269
5607	MITOGEN-ACTIVATED PROTEIN KINASE KINASE 5	MAP2K5	−3.44358		−2.68431
9064	MITOGEN-ACTIVATED PROTEIN KINASE KINASE KINASE 6	MAP3K6	−3.60211	−2.69142
6885	MITOGEN-ACTIVATED PROTEIN KINASE KINASE KINASE 7	MAP3K7	−4.37075	−2.60108	−3.08528
54799	MBT DOMAIN CONTAINING 1	MBTD1		−2.01418	−2.29198
92014	MITOCHONDRIAL CARRIER TRIPLE REPEAT 1	MCART1	−2.73059	−3.29002	−2.95126
4172	MCM3 MINICHROMOSOME MAINTENANCE DEFICIENT 3 (S. CEREVISIAE)	MCM3	−2.49008	−3.67504
28985	MALIGNANT T CELL AMPLIFIED SEQUENCE 1	MCTS1	−1.96517		−2.46353
9656	MEDIATOR OF DNA DAMAGE CHECKPOINT 1	MDC1	−2.65123		−2.46605
4197	ECOTROPIC VIRAL INTEGRATION SITE 1	MDS1	−4.91817	−4.36995	−3.49786
400569	SIMILAR TO HSPC296	MED11	−2.33766		−1.89359
80306	MEDIATOR OF RNA POLYMERASE II TRANSCRIPTION, SUBUNIT 28	MED28		−2.33833	−2.31198
	HOMOLOG (YEAST)
51003	MEDIATOR OF RNA POLYMERASE II TRANSCRIPTION, SUBUNIT 31	MED31	−2.04877	−3.62991
	HOMOLOG (YEAST)
4207	MADS BOX TRANSCRIPTION ENHANCER FACTOR 2, POLYPEPTIDE B	MEF2B	−5.35307	−4.30158	−1.89447
	(MYOCYTE ENHANCER FACTOR 2B)
1954	EGF-LIKE-DOMAIN, MULTIPLE 4	MEGF8	−2.25397	−2.14012	−3.97425
64747	MAJOR FACILITATOR SUPERFAMILY DOMAIN CONTAINING 1	MFSD1	−3.55784		−2.97033
84804	HYPOTHETICAL PROTEIN MGC11332	MFSD9	−2.2117		−2.04748
80772	HYPOTHETICAL PROTEIN MGC10334	MGC10334	−3.15307		−2.81389
92806	HYPOTHETICAL PROTEIN MGC13198	MGC16385	−3.67807	−2.73927	−5.34412
167359	HYPOTHETICAL PROTEIN MGC42105	MGC42105	−2.13288	−2.55735	−4.37954
401145	SIMILAR TO KIAA1680 PROTEIN	MGC48628	−2.65392	−1.9292
4259	MICROSOMAL GLUTATHIONE S-TRANSFERASE 3	MGST3	−3.23003	−2.59358	−2.15506
166968	HYPOTHETICAL PROTEIN	MIER3	−1.948	−2.33645	−2.37991
4323	MATRIX METALLOPEPTIDASE 14 (MEMBRANE-INSERTED)	MMP14		−2.50866	−2.79277
283385	MORN REPEAT CONTAINING 3	MORN3	−1.89433		−2.20862
758	METALLOPHOSPHOESTERASE DOMAIN CONTAINING 1	MPPED1		−2.74881	−1.94218
4360	MANNOSE RECEPTOR, C TYPE 1	MRC1	−2.64776		−3.15523
4361	MRE11 MEIOTIC RECOMBINATION 11 HOMOLOG A (S. CEREVISIAE)	MRE11A	−4.30835		−3.5375
64981	MITOCHONDRIAL RIBOSOMAL PROTEIN L34	MRPL34	−2.75412	−4.96395
341116	MEMBRANE-SPANNING 4-DOMAINS, SUBFAMILY A, MEMBER 10	MS4A10	−2.65751	−2.93441	−3.4722
4477	MICROSEMINOPROTEIN, BETA-	MSMB		−2.88215	−2.46353
4504	METALLOTHIONEIN 3 (GROWTH INHIBITORY FACTOR	MT3	−4.04357	−2.8508
	(NEUROTROPHIC))
8776	MYOTUBULARIN RELATED PROTEIN 1	MTMR1	−4.38585	−1.85331	−2.20333
136319	MYOTROPHIN	MTPN	−4.08597	−4.42856	−2.97718
4547	MICROSOMAL TRIGLYCERIDE TRANSFER PROTEIN	MTTP	−2.50767	−2.26107	−4.23045
57509	MITOCHONDRIAL TUMOR SUPPRESSOR 1	MTUS1		−3.12976	−2.58471
143662	MUCIN 15	MUC15	−2.19721	−2.12986
4589	MUCIN 7, SALIVARY	MUC7	−3.0859	−2.50545	−3.22334
4599	MYXOVIRUS (INFLUENZA VIRUS) RESISTANCE 1, INTERFERON-	MX1	−2.36526	−2.01844	−2.50688
	INDUCIBLE PROTEIN P78 (MOUSE)
343263	MYOSIN BINDING PROTEIN H-LIKE	MYBPHL	−2.91997		−1.87908
4641	MYOSIN IC	MYO1C	−3.75824	−2.04836
79829	HYPOTHETICAL PROTEIN FLJ13848	NAT11	−2.49128	−2.41016	−1.93249
26151	N-ACETYLTRANSFERASE 9	NAT9	−2.83485	−2.2593
284565	NEUROBLASTOMA BREAKPOINT FAMILY, MEMBER 14	NBPF15	−4.33412	−2.39879
83988	NEUROCALCIN DELTA	NCALD	−2.80549		−1.98663
4739	NEURAL PRECURSOR CELL EXPRESSED, DEVELOPMENTALLY DOWN-	NEDD9	−2.31196	−3.62758	−2.72499
	REGULATED 9
4751	NIMA (NEVER IN MITOSIS GENE A)-RELATED KINASE 2	NEK2	−2.9558	−2.43274	−2.95423
26012	NASAL EMBRYONIC LHRH FACTOR	NELF		−2.52297	−2.09509
4776	NUCLEAR FACTOR OF ACTIVATED T-CELLS, CYTOPLASMIC,	NFATC4	−3.61264	−3.21774
	CALCINEURIN-DEPENDENT 4
4778	NUCLEAR FACTOR (ERYTHROID-DERIVED 2), 45 KDA	NFE2		−2.12929	−2.10967
4802	NUCLEAR TRANSCRIPTION FACTOR Y, GAMMA	NFYC	−1.91075	−2.64669	−1.89951
159296	NK2 TRANSCRIPTION FACTOR HOMOLOG C (DROSOPHILA)	NKX2-3	−3.37969	−2.68783	−2.46978
51701	NEMO-LIKE KINASE	NLK	−3.33568	−1.88129	−2.20263
4829	NEUROMEDIN B RECEPTOR	NMBR	−2.78007	−3.06621
10201	NON-METASTATIC CELLS 6, PROTEIN EXPRESSED IN (NUCLEOSIDE-	NME6	−2.20441	−2.52635
	DIPHOSPHATE KINASE)
129521	NEUROMEDIN S	NMS	−5.61174	−3.74196	−2.58827
23530	NICOTINAMIDE NUCLEOTIDE TRANSHYDROGENASE	NNT	−3.34428	−3.40667
4838	NODAL HOMOLOG (MOUSE)	NODAL	−4.72064	−2.31463
27035	NADPH OXIDASE 1	NOX1	−2.99102		−1.9221
152519	NIPA-LIKE DOMAIN CONTAINING 1	NPAL1	−2.59477	−2.24146
190	NUCLEAR RECEPTOR SUBFAMILY 0, GROUP B, MEMBER 1	NR0B1	−2.82937	−2.00294
4929	NUCLEAR RECEPTOR SUBFAMILY 4, GROUP A, MEMBER 2	NR4A2	−4.44886	−5.87805	−2.9839
340371	NUCLEAR RECEPTOR BINDING PROTEIN 2	NRBP2		−2.79772	−2.53212
4898	NARDILYSIN (N-ARGININE DIBASIC CONVERTASE)	NRD1	−4.28791	−2.27774	−2.58532
83714	NUCLEAR RECEPTOR INTERACTING PROTEIN 2	NRIP2	−3.89044	−5.23847	−4.89818
22978	5′-NUCLEOTIDASE, CYTOSOLIC II	NT5C2		−2.93429	−3.38901
4908	NEUROTROPHIN 3	NTF3	−2.8015	−4.06468
4917	NETRIN 2-LIKE (CHICKEN)	NTN2L	−3.4598	−3.1736
4923	NEUROTENSIN RECEPTOR 1 (HIGH AFFINITY)	NTSR1	−4.10473	−3.49957
23620	NEUROTENSIN RECEPTOR 2	NTSR2	−3.90077	−4.85068
256281	NUDIX (NUCLEOSIDE DIPHOSPHATE LINKED MOIETY X)-TYPE MOTIF 14	NUDT14	−2.33744	−4.58128
51203	NUCLEOLAR AND SPINDLE ASSOCIATED PROTEIN 1	NUSAP1	−3.24207	−2.43638
56000	NUCLEAR RNA EXPORT FACTOR 3	NXF3	−4.03665	−4.20158	−2.41973
55916	NUCLEAR TRANSPORT FACTOR 2-LIKE EXPORT FACTOR 2	NXT2	−2.78623	−2.45246
220323	OAF HOMOLOG (DROSOPHILA)	OAF	−3.31157	−1.91214
51686	ORNITHINE DECARBOXYLASE ANTIZYME 3	OAZ3	−4.98126		−3.05747
4952	OCULOCEREBRORENAL SYNDROME OF LOWE	OCRL	−3.56185		−3.44429
4957	OUTER DENSE FIBER OF SPERM TAILS 2	ODF2	−1.98896	−2.06861
169611	OLFACTOMEDIN-LIKE 2A	OLFML2A	−2.18173	−2.26417
10133	OPTINEURIN	OPTN	−3.13476	−2.18057
138802	OLFACTORY RECEPTOR, FAMILY 13, SUBFAMILY C, MEMBER 8	OR13C8		−4.09071	−2.51101
4991	OLFACTORY RECEPTOR, FAMILY 1, SUBFAMILY D, MEMBER 2	OR1D2	−3.16865	−3.64961	−3.4325
347168	OLFACTORY RECEPTOR, FAMILY 1, SUBFAMILY J, MEMBER 1	OR1J1	−2.01037	−2.05408	−3.5605
392392	OLFACTORY RECEPTOR, FAMILY 1, SUBFAMILY K, MEMBER 1	OR1K1	−3.90467	−2.16026
144125	OLFACTORY RECEPTOR, FAMILY 2, SUBFAMILY AG, MEMBER 1	OR2AG1	−1.90182	−1.93355
341152	OLFACTORY RECEPTOR, FAMILY 2, SUBFAMILY AT, MEMBER 4	OR2AT4	−2.69945	−2.16615
81469	OLFACTORY RECEPTOR, FAMILY 2, SUBFAMILY G, MEMBER 3	OR2G3	−1.96137		−2.16489
391194	OLFACTORY RECEPTOR, FAMILY 2, SUBFAMILY M, MEMBER 2	OR2M2		−2.41789	−1.98612
26245	OLFACTORY RECEPTOR, FAMILY 2, SUBFAMILY M, MEMBER 4	OR2M4		−2.72931	−2.1423
127069	OLFACTORY RECEPTOR, FAMILY 2, SUBFAMILY T, MEMBER 10	OR2T10	−3.65767	−2.24491
284383	OLFACTORY RECEPTOR, FAMILY 2, SUBFAMILY Z, MEMBER 1	OR2Z1	−3.07678	−4.72263
219429	OLFACTORY RECEPTOR, FAMILY 4, SUBFAMILY C, MEMBER 11	OR4C11	−2.64811	−2.09856
79317	OLFACTORY RECEPTOR, FAMILY 4, SUBFAMILY K, MEMBER 5	OR4K5	−2.60077	−1.91433
81300	OLFACTORY RECEPTOR, FAMILY 4, SUBFAMILY P, MEMBER 4	OR4P4	−4.49028	−2.64832	−2.25624
256148	OLFACTORY RECEPTOR, FAMILY 4, SUBFAMILY S, MEMBER 1	OR4S1	−2.26199	−1.94054
196335	OLFACTORY RECEPTOR, FAMILY 56, SUBFAMILY B, MEMBER 4	OR56B4	−5.35355	−1.91302
403274	OLFACTORY RECEPTOR, FAMILY 5, SUBFAMILY H, MEMBER 15	OR5H15	−6.03036		−2.24129
120065	OLFACTORY RECEPTOR, FAMILY 5, SUBFAMILY P, MEMBER 2	OR5P2	−2.71333		−2.64519
390154	OLFACTORY RECEPTOR, FAMILY 5, SUBFAMILY T, MEMBER 3	OR5T3	−1.89065		−2.19858
391114	OLFACTORY RECEPTOR, FAMILY 6, SUBFAMILY K, MEMBER 3	OR6K3		−2.47585	−2.64133
10956	AMPLIFIED IN OSTEOSARCOMA	OS9		−1.88867	−2.8692
55074	OXIDATION RESISTANCE 1	OXR1	−2.56001	−3.04851
54995	3-OXOACYL-ACP SYNTHASE, MITOCHONDRIAL	OXSM		−2.55496	−2.11533
125988	QIL1 PROTEIN	P117	−3.33321		−1.88138
23241	PHOSPHOFURIN ACIDIC CLUSTER SORTING PROTEIN 2	PACS2	−2.36166		−2.96823
5050	PLATELET-ACTIVATING FACTOR ACETYLHYDROLASE, ISOFORM IB,	PAFAH1B3	−1.90165		−2.03995
	GAMMA SUBUNIT 29 KDA
23022	PALLADIN, CYTOSKELETAL ASSOCIATED PROTEIN	PALLD	−1.87671		−2.14591
10914	POLY(A) POLYMERASE ALPHA	PAPOLA	−2.08643	−4.05228
124222	PROGESTIN AND ADIPOQ RECEPTOR FAMILY MEMBER IV	PAQR4	−2.39582		−3.38779
142	POLY (ADP-RIBOSE) POLYMERASE FAMILY, MEMBER 1	PARP1		−1.92528	−1.90532
143	POLY (ADP-RIBOSE) POLYMERASE FAMILY, MEMBER 4	PARP4		−2.40935	−2.06206
27253	PROTOCADHERIN 17	PCDH17		−2.21436	−3.0782
57526	PROTOCADHERIN 19	PCDH19	−3.15028	−2.86698
56132	PROTOCADHERIN BETA 3	PCDHB3		−2.6344	−2.62729
10336	POLYCOMB GROUP RING FINGER 3	PCGF3	−3.03471		−1.89414
84333	POLYCOMB GROUP RING FINGER 5	PCGF5	−2.773	−1.96929	−4.09651
55795	HYPOTHETICAL PROTEIN FLJ11305	PCID2		−1.98532	−2.32044
5046	PROPROTEIN CONVERTASE SUBTILISIN/KEXIN TYPE 6	PCSK6	−2.1397	−3.10903
58488	PHOSPHATIDYLCHOLINE TRANSFER PROTEIN	PCTP	−6.35862	−3.17278	−1.87497
5161	PYRUVATE DEHYDROGENASE (LIPOAMIDE) ALPHA 2	PDHA2	−2.027	−3.19203
5166	PYRUVATE DEHYDROGENASE KINASE, ISOZYME 4	PDK4	−2.08325		−1.86973
9260	PDZ AND LIM DOMAIN 7 (ENIGMA)	PDLIM7		−2.14491	−1.88778
57546	PYRUVATE DEHYDROGENASE PHOSPHATASE ISOENZYME 2	PDP2	−2.63359	−2.24757
3651	INSULIN PROMOTER FACTOR 1, HOMEODOMAIN TRANSCRIPTION	PDX1		−2.14427	−2.43953
	FACTOR
51248	PDZ DOMAIN CONTAINING 11	PDZD11	−2.05074	−2.52156
5179	PROENKEPHALIN	PENK	−1.99554	−2.52421
64065	PERP, TP53 APOPTOSIS EFFECTOR	PERP		−3.04715	−1.86263
5210	6-PHOSPHOFRUCTO-2-KINASE/FRUCTOSE-2,6-BIPHOSPHATASE 4	PFKFB4	−2.80393		−2.73894
80055	GPI DEACYLASE	PGAP1	−2.47747	−4.57794	−2.70667
267004	EXCISION REPAIR CROSS-COMPLEMENTING RODENT REPAIR	PGBD3	−4.20479		−1.88145
	DEFICIENCY, COMPLEMENTATION GROUP 6
57115	PEPTIDOGLYCAN RECOGNITION PROTEIN 4	PGLYRP4		−1.99135	−1.85699
84680	1-AMINOCYCLOPROPANE-1-CARBOXYLATE SYNTHASE	PHACS	−3.19355	−2.15665
84295	PHD FINGER PROTEIN 6	PHF6	−2.28931	−2.38237
22822	PLECKSTRIN HOMOLOGY-LIKE DOMAIN, FAMILY A, MEMBER 1	PHLDA1	−2.63006		−2.53575
221476	PEPTIDASE INHIBITOR 16	PI16	−2.17842		−2.47888
5277	PHOSPHATIDYLINOSITOL GLYCAN, CLASS A (PAROXYSMAL	PIGA	−2.7267		−1.97114
	NOCTURNAL HEMOGLOBINURIA)
5289	PHOSPHOINOSITIDE-3-KINASE, CLASS 3	PIK3C3	−3.02798	−2.16509
65018	PTEN INDUCED PUTATIVE KINASE 1	PINK1	−2.65238	−6.72753	−4.89433
54984	PIN2-INTERACTING PROTEIN 1	PINX1	−2.13509	−2.25069
8395	PHOSPHATIDYLINOSITOL-4-PHOSPHATE 5-KINASE, TYPE I, BETA	PIP5K1B	−2.49714	−3.11396	−2.30031
23761	PHOSPHATIDYLSERINE DECARBOXYLASE	PISD	−2.77933	−3.93785
5314	POLYCYSTIC KIDNEY AND HEPATIC DISEASE 1 (AUTOSOMAL	PKHD1	−1.95223	−2.59701
	RECESSIVE)
5569	PROTEIN KINASE (CAMP-DEPENDENT, CATALYTIC) INHIBITOR ALPHA	PKIA	−3.57576	−2.04651
29941	PROTEIN KINASE N3	PKN3		−2.10742	−2.15859
5318	PLAKOPHILIN 2	PKP2	−2.89785	−2.30137
283748	PHOSPHOLIPASE A2, GROUP IVD (CYTOSOLIC)	PLA2G4D	−1.96541		2.51478
5322	PHOSPHOLIPASE A2, GROUP V	PLA2G5	−2.68114		−2.08402
5326	PLEIOMORPHIC ADENOMA GENE-LIKE 2	PLAGL2		−4.21387	−2.5165
5327	PLASMINOGEN ACTIVATOR, TISSUE	PLAT		−3.25981	−3.48747
5332	PHOSPHOLIPASE C, BETA 4	PLCB4	−2.49701	−2.10791
257068	PHOSPHATIDYLINOSITOL-SPECIFIC PHOSPHOLIPASE C, X DOMAIN	PLCXD2	−3.6423	−2.74439
	CONTAINING 2
54477	PLECKSTRIN HOMOLOGY DOMAIN CONTAINING, FAMILY A MEMBER 5	PLEKHA5	−2.2379	−2.30195
58473	PLECKSTRIN HOMOLOGY DOMAIN CONTAINING, FAMILY B (EVECTINS)	PLEKHB1		−2.58955	−2.51903
	MEMBER 1
11284	POLYNUCLEOTIDE KINASE 3′-PHOSPHATASE	PNKP	−1.97244	−2.18221	−2.14383
79883	HYPOTHETICAL PROTEIN FLJ23447	PODNL1	−4.69966	−2.85234
79001	VITAMIN K EPOXIDE REDUCTASE COMPLEX, SUBUNIT 1	POL3S		−2.15713	−2.25578
23649	POLYMERASE (DNA DIRECTED), ALPHA 2 (70 KD SUBUNIT)	POLA2	−2.98897	−2.75611
54107	POLYMERASE (DNA DIRECTED), EPSILON 3 (P17 SUBUNIT)	POLE3	−3.28031	−2.07165	−2.89831
94026	POM121 MEMBRANE GLYCOPROTEIN-LIKE 2 (RAT)	POM121L2	−4.64585		−2.05884
29954	PROTEIN-O-MANNOSYLTRANSFERASE 2	POMT2	−2.84072		−1.962
5446	PARAOXONASE 3	PON3	−4.70654	−2.34705
10940	PROCESSING OF PRECURSOR 1, RIBONUCLEASE P/MRP SUBUNIT	POP1	−4.49314	−2.46298	−1.97797
	(S. CEREVISIAE)
25833	POU DOMAIN, CLASS 2, TRANSCRIPTION FACTOR 3	POU2F3	−3.34571	−2.41621	−2.29749
8612	PHOSPHATIDIC ACID PHOSPHATASE TYPE 2C	PPAP2C	−2.95992	−2.07329
84814	PHOSPHATIDIC ACID PHOSPHATASE TYPE 2 DOMAIN CONTAINING 3	PPAPDC3	−1.95667		−1.96121
8499	PROTEIN TYROSINE PHOSPHATASE, RECEPTOR TYPE, F POLYPEPTIDE	PPFIA2		−2.04642	−2.65517
	(PTPRF), INTERACTING PROTEIN (LIPRIN), ALPHA 2
9360	PEPTIDYLPROLYL ISOMERASE G (CYCLOPHILIN G)	PPIG	−3.51287		−2.43114
5498	PROTOPORPHYRINOGEN OXIDASE	PPOX	−1.91497	−2.02656
4659	PROTEIN PHOSPHATASE 1, REGULATORY (INHIBITOR) SUBUNIT 12A	PPP1R12A	−2.86581		−2.07108
79660	PROTEIN PHOSPHATASE 1, REGULATORY (INHIBITOR) SUBUNIT 3B	PPP1R3B	−3.05123	−1.9493
5515	PROTEIN PHOSPHATASE 2 (FORMERLY 2A), CATALYTIC SUBUNIT,	PPP2CB	−5.1951	−3.82763
	ALPHA ISOFORM
55012	CHROMOSOME 14 OPEN READING FRAME 10	PPP2R3C	−1.90047	−2.87177	−1.99879
5525	PROTEIN PHOSPHATASE 2, REGULATORY SUBUNIT B (B56), ALPHA	PPP2R5A	−1.85237	−1.88221
	ISOFORM
23082	PEROXISOME PROLIFERATIVE ACTIVATED RECEPTOR, GAMMA,	PPRC1	−3.33778		−2.53909
	COACTIVATOR-RELATED 1
65121	PRAME FAMILY MEMBER 1	PRAMEF12	−1.95611	−2.12128
9055	PROTEIN REGULATOR OF CYTOKINESIS 1	PRC1	−4.69961	−2.34331	−2.33972
5551	PERFORIN 1 (PORE FORMING PROTEIN)	PRF1	−3.11112	−4.12973	−3.64376
5562	PROTEIN KINASE, AMP-ACTIVATED, ALPHA 1 CATALYTIC SUBUNIT	PRKAA1		−2.42841	−3.18606
5568	PROTEIN KINASE, CAMP-DEPENDENT, CATALYTIC, GAMMA	PRKACG		−2.27403	−2.2523
56341	PROTEIN ARGININE METHYLTRANSFERASE 8	PRMT8	−2.53607		−2.38768
5626	PROPHET OF PIT1, PAIRED-LIKE HOMEODOMAIN TRANSCRIPTION	PROP1		−3.17865	−1.91607
	FACTOR
51334	MESENCHYMAL STEM CELL PROTEIN DSC54	PRR16	−1.89517	−2.22191
10279	PROTEASE, SERINE, 16 (THYMUS)	PRSS16	−2.13449	−2.63428	−3.12227
400668	PROTEASE, SERINE-LIKE 1	PRSSL1		−1.87807	−2.17207
57716	PERIAXIN	PRX		−3.7748	−2.19862
9595	PLECKSTRIN HOMOLOGY, SEC7 AND COILED-COIL DOMAINS, BINDING	PSCDBP		−2.27685	−2.20532
	PROTEIN
5681	PROTEIN SERINE KINASE H1	PSKH1	−2.63033		−2.63583
139411	PATCHED DOMAIN CONTAINING 1	PTCHD1	−2.0655	−2.79843
81490	PHOSPHATIDYLSERINE SYNTHASE 2	PTDSS2		−2.19403	−1.9922
11099	PROTEIN TYROSINE PHOSPHATASE, NON-RECEPTOR TYPE 21	PTPN21	−2.49387	−2.04587
5787	PROTEIN TYROSINE PHOSPHATASE, RECEPTOR TYPE, B	PTPRB	−2.9585	−2.07646
5794	PROTEIN TYROSINE PHOSPHATASE, RECEPTOR TYPE, H	PTPRH	−2.91132	−2.41963
5801	PROTEIN TYROSINE PHOSPHATASE, RECEPTOR TYPE, R	PTPRR		−2.05897	−2.10197
5814	PURINE-RICH ELEMENT BINDING PROTEIN B	PURB	−3.03529	−2.0988
79912	HYPOTHETICAL PROTEIN FLJ22028	PYROXD1	−2.08054	−3.59087	−3.50611
5697	PEPTIDE YY	PYY	−3.01242	−1.89304
9727	RAB11 FAMILY INTERACTING PROTEIN 3 (CLASS II)	RAB11FIP3	−1.93666	−2.06647
401409	GTP-BINDING PROTEIN RAB19B	RAB19	−4.16718	−2.50667
11021	RAB35, MEMBER RAS ONCOGENE FAMILY	RAB35	−3.72915	−2.10077
116442	RAB39B, MEMBER RAS ONCOGENE FAMILY	RAB39B	−3.40849	−2.77826	−2.55766
5867	RAB4A, MEMBER RAS ONCOGENE FAMILY	RAB4A		−2.23824	−2.38798
53916	RAB4B, MEMBER RAS ONCOGENE FAMILY	RAB4B	−2.62903	−2.16647
5885	RAD21 HOMOLOG (S. POMBE)	RAD21		−3.13061	−2.07017
23132	RAD54-LIKE 2 (S. CEREVISIAE)	RAD54L2		−2.48403	−3.18984
5883	RAD9 HOMOLOG A (S. POMBE)	RAD9A	−1.98876	−2.71587	−2.70529
22913	RNA BINDING PROTEIN, AUTOANTIGENIC (HNRNP-ASSOCIATED WITH	RALY		−2.31833	−2.07084
	LETHAL YELLOW HOMOLOG (MOUSE))
26953	RAN BINDING PROTEIN 6	RANBP6	−2.83259		−1.8949
5906	RAP1A, MEMBER OF RAS ONCOGENE FAMILY	RAP1A	−2.15637	−2.59656
5920	RETINOIC ACID RECEPTOR RESPONDER (TAZAROTENE INDUCED) 3	RARRES3	−3.59647	−2.88117	−3.01349
25780	RAS GUANYL RELEASING PROTEIN 3 (CALCIUM AND DAG-REGULATED)	RASGRP3		−2.68079	−2.15504
64080	RIBOKINASE	RBKS	−2.70105	−3.24448
54033	RNA BINDING MOTIF PROTEIN 11	RBM11		−3.16829	−1.94764
166863	HYPOTHETICAL PROTEIN MGC27016	RBM46	−2.66251		−2.8393
23543	RNA BINDING MOTIF PROTEIN 9	RBM9	−4.51469	−2.6588
83758	RETINOL BINDING PROTEIN 5, CELLULAR	RBP5	−2.07092	−2.56384	−2.33793
11317	RECOMBINING BINDING PROTEIN SUPPRESSOR OF HAIRLESS	RBPJL	−3.4381	−2.8404
	(DROSOPHILA)-LIKE
348093	RNA BINDING PROTEIN WITH MULTIPLE SPLICING 2	RBPMS2	−4.50172	−3.38917	−2.0127
5957	RECOVERIN	RCVRN		−2.95204	−3.18587
7936	RD RNA BINDING PROTEIN	RDBP	−2.08726	−2.16704
5962	RADIXIN	RDX	−2.37925	−2.62954
51308	RECEPTOR ACCESSORY PROTEIN 2	REEP2	−2.85997	−1.96118
56729	RESISTIN	RETN	−1.93284	−1.90428	−2.73758
55312	RIBOFLAVIN KINASE	RFK		−1.90106	−2.43082
442247	SIMILAR TO RET FINGER PROTEIN-LIKE 1	RFPL4B	−3.39753	−2.28722
93587	RNA (GUANINE-9-) METHYLTRANSFERASE DOMAIN CONTAINING 2	RG9MTD2	−3.08085	−2.39402
5996	REGULATOR OF G-PROTEIN SIGNALLING 1	RGS1	−2.6945	−4.44163
26166	REGULATOR OF G-PROTEIN SIGNALLING 22	RGS22	−1.99317	−2.56763
121268	RAS HOMOLOG ENRICHED IN BRAIN LIKE 1	RHEBL1	−3.3124	−2.59878
54509	RAS HOMOLOG GENE FAMILY, MEMBER F (IN FILOPODIA)	RHOF	−1.99428		−2.60677
399	RAS HOMOLOG GENE FAMILY, MEMBER H	RHOH	−2.87756	−2.60693
25778	RECEPTOR INTERACTING PROTEIN KINASE 5	RIPK5	−2.16914		−3.07716
6039	RIBONUCLEASE, RNASE A FAMILY, K6	RNASE6	−3.83481	−3.37285
140432	RING FINGER PROTEIN 113B	RNF113B		−2.17001	−2.80417
79845	RING FINGER PROTEIN 122	RNF122	−2.16016	−3.10137
54546	RING FINGER PROTEIN 186	RNF186	−1.95557		−2.34727
6050	RIBONUCLEASE/ANGIOGENIN INHIBITOR 1	RNH1		−1.85151	−2.2123
10921	RNA BINDING PROTEIN S1, SERINE-RICH DOMAIN	RNPS1	−2.23163	−2.8619
10556	RIBONUCLEASE P/MRP 30 KDA SUBUNIT	RPP30	−2.7066	−3.00534	−2.13315
56261	HYPOTHETICAL PROTEIN KIAA1434	RPS18P1	−2.85418	−2.69983
91582	RIBOSOMAL PROTEIN S19 BINDING PROTEIN 1	RPS19BP1	−2.21276	−2.78638
8986	RIBOSOMAL PROTEIN S6 KINASE, 90 KDA, POLYPEPTIDE 4	RPS6KA4	−1.93966	−2.88556
9136	RNA, U3 SMALL NUCLEOLAR INTERACTING PROTEIN 2	RRP9	−2.34514		−1.86564
84870	R-SPONDIN 3 HOMOLOG (XENOPUS LAEVIS)	RSPO3	−2.13463	−2.68373
146760	RETICULON 4 RECEPTOR-LIKE 1	RTN4RL1		−2.07964	−2.34172
146923	RUN DOMAIN CONTAINING 1	RUNDC1	−2.18962	−2.04771
154661	RAP2-BINDING PROTEIN 9	RUNDC3B		−2.9338	−3.37057
862	RUNT-RELATED TRANSCRIPTION FACTOR 1; TRANSLOCATED TO, 1	RUNX1T1	−2.12615	−2.27231
	(CYCLIN D-RELATED)
23429	RING1 AND YY1 BINDING PROTEIN	RYBP		−2.02569	−2.10047
645922	SIMILAR TO S100 CALCIUM BINDING PROTEIN A7-LIKE 1	S100A7L2	−3.42865	−2.28363	−2.64235
6285	S100 CALCIUM BINDING PROTEIN, BETA (NEURAL)	S100B	−2.2964		−3.721
113174	SERUM AMYLOID A-LIKE 1	SAAL1	−3.11984		−3.5017
27164	SAL-LIKE 3 (DROSOPHILA)	SALL3		−2.34564	−2.04659
344658	STERILE ALPHA MOTIF DOMAIN CONTAINING 7	SAMD7		−2.51076	−2.13509
55291	CHROMOSOME 11 OPEN READING FRAME 23	SAPS3		−2.90252	−1.8513
163786	SPINDLE ASSEMBLY 6 HOMOLOG (C. ELEGANS)	SASS6		−1.97416	−2.54308
23314	SATB FAMILY MEMBER 2	SATB2		−2.85258	−2.07986
23256	SEC1 FAMILY DOMAIN CONTAINING 1	SCFD1	−2.41869	−3.0156
7857	SECRETOGRANIN II (CHROMOGRANIN C)	SCG2	−2.43482		−2.45974
6332	SODIUM CHANNEL, VOLTAGE-GATED, TYPE VII, ALPHA	SCN7A	−4.12895	−2.66506
51246	SCOTIN	SCOTIN	−4.07991	−2.10949
6343	SECRETIN	SCT	−4.44056	−2.51525
80274	SIGNAL PEPTIDE, CUB DOMAIN, EGF-LIKE 1	SCUBE1		−2.61249	−2.47153
9255	SMALL INDUCIBLE CYTOKINE SUBFAMILY E, MEMBER 1 (ENDOTHELIAL	SCYE1	−2.36951	−2.70046
	MONOCYTE-ACTIVATING)
9672	SYNDECAN 3 (N-SYNDECAN)	SDC3	−2.91628		−2.81579
29927	SEC61 ALPHA 1 SUBUNIT (S. CEREVISIAE)	SEC61A1	−1.88201	−3.10851
55176	SEC61 ALPHA 2 SUBUNIT (S. CEREVISIAE)	SEC61A2		−1.90727	−2.15394
10952	SEC61 BETA SUBUNIT	SEC61B		−1.90117	−2.31682
6404	SELECTIN P LIGAND	SELPLG	−2.09831	−3.20512
348303	SELENOPROTEIN V	SELV		−3.17995	−2.28173
9037	SEMA DOMAIN, SEVEN THROMBOSPONDIN REPEATS (TYPE 1 AND	SEMA5A		−2.55799	−2.73749
	TYPE 1-LIKE), TRANSMEMBRANE DOMAIN (TM) AND SHORT
	CYTOPLASMIC DOMAIN, (SEMAPHORIN) 5A
10500	SEMA DOMAIN, TRANSMEMBRANE DOMAIN (TM), AND CYTOPLASMIC	SEMA6C	−3.43802	−1.91085
	DOMAIN, (SEMAPHORIN) 6C
26054	SUMO1/SENTRIN SPECIFIC PEPTIDASE 6	SENP6	−2.12559	−4.92084
1992	SERPIN PEPTIDASE INHIBITOR, CLADE B (OVALBUMIN), MEMBER 1	SERPINB1	−3.26772	−2.86627	−2.8311
462	SERPIN PEPTIDASE INHIBITOR, CLADE C (ANTITHROMBIN), MEMBER 1	SERPINC1		−2.62448	−1.95094
143686	SESTRIN 3	SESN3	−2.41865	−2.54664
387893	SET DOMAIN CONTAINING (LYSINE METHYLTRANSFERASE) 8	SETD8	−2.21435	−2.14105
9295	SPLICING FACTOR, ARGININE/SERINE-RICH 11	SFRS11	−3.46224	−2.01065
25957	CHROMOSOME 6 OPEN READING FRAME 111	SFRS18	−2.8846	−2.66063
6457	SH3-DOMAIN GRB2-LIKE 3	SH3GL3	−2.09546		−1.90088
22941	SH3 AND MULTIPLE ANKYRIN REPEAT DOMAINS 2	SHANK2	−1.85617	−2.35166
6462	SEX HORMONE-BINDING GLOBULIN	SHBG		−2.12767	−3.0562
134549	APICAL PROTEIN 2	SHROOM1	−2.68342	−2.30012
25942	SIN3 HOMOLOG A, TRANSCRIPTION REGULATOR (YEAST)	SIN3A	−2.50484		−1.91435
23094	SIGNAL-INDUCED PROLIFERATION-ASSOCIATED 1 LIKE 3	SIPA1L3		−2.00188	−1.98104
6498	SKI-LIKE	SKIL	−2.52428	−2.56457	−2.61359
84174	SRC-LIKE-ADAPTOR 2	SLA2		−3.20957	−2.37006
4891	SOLUTE CARRIER FAMILY 11 (PROTON-COUPLED DIVALENT METAL ION	SLC11A2	−2.54813	−3.59077
	TRANSPORTERS), MEMBER 2
6560	SOLUTE CARRIER FAMILY 12 (POTASSIUM/CHLORIDE TRANSPORTERS),	SLC12A4	−1.91482	−2.6526	−3.08265
	MEMBER 4
117247	HYPOTHETICAL PROTEIN PRO0813	SLC16A10	−2.62914	−2.48375
55356	SOLUTE CARRIER FAMILY 22 (ORGANIC CATION TRANSPORTER),	SLC22A15	−2.86114	−2.176
	MEMBER 15
8604	SOLUTE CARRIER FAMILY 25 (MITOCHONDRIAL CARRIER, ARALAR),	SLC25A12	−3.02631		−2.05383
	MEMBER 12
115286	SOLUTE CARRIER FAMILY 25, MEMBER 26	SLC25A26	−4.64485	−4.72509
291	SOLUTE CARRIER FAMILY 25 (MITOCHONDRIAL CARRIER; ADENINE	SLC25A4	−2.38678	−2.19747
	NUCLEOTIDE TRANSLOCATOR), MEMBER 4
11001	FATTY-ACID-COENZYME A LIGASE, VERY LONG-CHAIN 1	SLC27A2	−2.29279		−2.10208
11000	SOLUTE CARRIER FAMILY 27 (FATTY ACID TRANSPORTER), MEMBER 3	SLC27A3		−1.86814	−2.04285
64078	SOLUTE CARRIER FAMILY 28 (SODIUM-COUPLED NUCLEOSIDE	SLC28A3	−2.72258		−2.00847
	TRANSPORTER), MEMBER 3
54733	SOLUTE CARRIER FAMILY 35, MEMBER F2	SLC35F2	−3.54646	−2.37764
23446	SOLUTE CARRIER FAMILY 44, MEMBER 1	SLC44A1	−2.74664	−2.60586
9152	SOLUTE CARRIER FAMILY 6 (NEUROTRANSMITTER TRANSPORTER,	SLC6A5	−3.13789		−2.174
	GLYCINE), MEMBER 5
6550	SOLUTE CARRIER FAMILY 9 (SODIUM/HYDROGEN EXCHANGER),	SLC9A3	−2.10203	−2.83787
	MEMBER 3
9351	SOLUTE CARRIER FAMILY 9 (SODIUM/HYDROGEN EXCHANGER),	SLC9A3R2		−1.99031	−2.77691
	MEMBER 3 REGULATOR 2
4088	SMAD, MOTHERS AGAINST DPP HOMOLOG 3 (DROSOPHILA)	SMAD3	−2.1217		−1.97539
60682	STROMAL MEMBRANE-ASSOCIATED PROTEIN 1	SMAP1	−4.06345		−2.32987
6603	SWI/SNF RELATED, MATRIX ASSOCIATED, ACTIN DEPENDENT	SMARCD2		−5.85968	−2.70953
	REGULATOR OF CHROMATIN, SUBFAMILY D, MEMBER 2
79677	SMC6 STRUCTURAL MAINTENANCE OF CHROMOSOMES 6-LIKE 1	SMC6		−2.24287	−3.68918
	(YEAST)
6609	SPHINGOMYELIN PHOSPHODIESTERASE 1, ACID LYSOSOMAL (ACID	SMPD1	−4.41417	−3.32313
	SPHINGOMYELINASE)
6525	SMOOTHELIN	SMTN	−1.97754	−4.5857
9751	SYNTAPHILIN	SNPH	−2.19133	−3.77141
6629	SMALL NUCLEAR RIBONUCLEOPROTEIN POLYPEPTIDE B”	SNRPB2	−2.3241	−2.37559
54212	SYNTROPHIN, GAMMA 1	SNTG1	−2.50921	−2.49363
29916	SORTING NEXIN 11	SNX11	−2.01535	−1.93444
23161	SORTING NEXIN 13	SNX13	−2.0599	−2.51066
6652	SORBITOL DEHYDROGENASE	SORD	−3.55463	−2.14119	−1.97628
6655	SON OF SEVENLESS HOMOLOG 2 (DROSOPHILA)	SOS2	−2.82991	−2.64977
9580	SRY (SEX DETERMINING REGION Y)-BOX 13	SOX13	−3.5731	−3.49442
6667	SP1 TRANSCRIPTION FACTOR	SP1	−2.90651	−3.43006
10615	SPERM ASSOCIATED ANTIGEN 5	SPAG5	−4.34344	−2.63317
80309	SPHK1 (SPHINGOSINE KINASE TYPE 1) INTERACTING PROTEIN	SPHKAP	−2.35706	−3.18283
643394	SIMILAR TO ESOPHAGUS CANCER-RELATED GENE 2 PROTEIN	SPINK9	−3.44822	−3.26333
	PRECURSOR (ECRG-2)
83985	SPINSTER	SPNS1	−2.93729	−2.13496	−3.33721
201305	HYPOTHETICAL PROTEIN MGC29671	SPNS3	−2.26795	−3.47607
6695	SPARC/OSTEONECTIN, CWCV AND KAZAL-LIKE DOMAINS	SPOCK1		−5.11991	−3.69976
	PROTEOGLYCAN (TESTICAN) 1
10418	SPONDIN 1, EXTRACELLULAR MATRIX PROTEIN	SPON1	−2.5145	−3.43213	−2.98118
6720	STEROL REGULATORY ELEMENT BINDING TRANSCRIPTION FACTOR 1	SREBF1	−2.05248	−1.98817
51188	SYNOVIAL SARCOMA TRANSLOCATION GENE ON CHROMOSOME 18-	SS18L2	−2.55102	−4.97016
	LIKE 2
23648	SINGLE STRANDED DNA BINDING PROTEIN 3	SSBP3	−1.90861	−3.22848
23145	SCO-SPONDIN HOMOLOG (BOS TAURUS)	SSPO	−3.57048	−2.81785	−4.46215
6745	SIGNAL SEQUENCE RECEPTOR, ALPHA (TRANSLOCON-ASSOCIATED	SSR1	−2.8973	−3.21686	−3.23373
	PROTEIN ALPHA)
9705	SUPPRESSION OF TUMORIGENICITY 18 (BREAST CARCINOMA) (ZINC	ST18	−3.4941		−3.10471
	FINGER PROTEIN)
29906	ST8 ALPHA-N-ACETYL-NEURAMINIDE ALPHA-2,8-SIALYLTRANSFERASE 5	ST8SIA5	−3.72604		−2.45803
27067	STAUFEN, RNA BINDING PROTEIN, HOMOLOG 2 (DROSOPHILA)	STAU2	−1.94355	−4.02935
8576	SERINE/THREONINE KINASE 16	STK16	−1.97744		−2.08507
29888	STRIATIN, CALMODULIN BINDING PROTEIN 4	STRN4	−4.29577	−3.06848
29091	SYNTAXIN BINDING PROTEIN 6 (AMISYN)	STXBP6		−2.09798	−2.18455
51684	SUPPRESSOR OF FUSED HOMOLOG (DROSOPHILA)	SUFU	−2.03606	−3.28222	−2.11306
285362	SULFATASE MODIFYING FACTOR 1	SUMF1	−3.73367	−2.48341
6836	SURFEIT 4	SURF4	−1.95495	−5.84907
23546	SYNAPTOGYRIN 4	SYNGR4	−2.93405		−1.89975
171024	SYNAPTOPODIN 2	SYNPO2	−3.58817		−3.00192
23208	SYNAPTOTAGMIN XI	SYT11	−3.85581	−4.17378
83849	SYNAPTOTAGMIN XV	SYT15	−3.73035	−3.0071	−2.76884
83851	SYNAPTOTAGMIN XVI	SYT16		−2.98035	−1.87882
90019	SYNAPTOTAGMIN VIII	SYT8	−3.26196	−1.94023
134864	TRACE AMINE ASSOCIATED RECEPTOR 1	TAAR1		−2.4933	−2.37326
9287	TRACE AMINE ASSOCIATED RECEPTOR 2	TAAR2	−2.07314	−2.94412	−2.55597
117143	TRANSCRIPTIONAL ADAPTOR 1 (HFI1 HOMOLOG, YEAST)-LIKE	TADA1L	−3.58325	−4.24081
10474	TRANSCRIPTIONAL ADAPTOR 3 (NGG1 HOMOLOG, YEAST)-LIKE	TADA3L	−3.69376	−2.67301	−2.67191
6883	TAF12 RNA POLYMERASE II, TATA BOX BINDING PROTEIN (TBP)-	TAF12	−3.506	−3.03956	−3.59767
	ASSOCIATED FACTOR, 20 KDA
8148	TAF15 RNA POLYMERASE II, TATA BOX BINDING PROTEIN (TBP)-	TAF15	−5.00259	−2.99249
	ASSOCIATED FACTOR, 68 KDA
389932	ALDO-KETO REDUCTASE, TRUNCATED	TAKR	−3.01732	−3.14658
6888	TRANSALDOLASE 1	TALDO1	−2.93536	−1.951
84807	T-CELL ACTIVATION NFKB-LIKE PROTEIN	TA-		−2.0215	−2.34425
		NFKBH
374403	TBC1 DOMAIN FAMILY, MEMBER 10C	TBC1D10C	−3.33335		−2.33856
23102	TBC1 DOMAIN FAMILY, MEMBER 2B	TBC1D2B	−2.38151	−3.63115	−2.23145
79718	TRANSDUCIN (BETA)-LIKE 1X-LINKED RECEPTOR 1	TBL1XR1		−1.98849	−2.93411
6926	T-BOX 3 (ULNAR MAMMARY SYNDROME)	TBX3	−2.84324	−2.55287	−2.1923
54103	HYPOTHETICAL PROTEIN LOC54103	TCAG7.1314		−1.97256	−3.65763
56849	TRANSCRIPTION ELONGATION FACTOR A (SII)-LIKE 7	TCEAL7	−4.02826	−2.17249
6921	TRANSCRIPTION ELONGATION FACTOR B (SIII), POLYPEPTIDE 1 (15 KDA,	TCEB1	−3.56259	−2.87088	−2.80807
	ELONGIN C)
9623	T-CELL LEUKEMIA/LYMPHOMA 1B	TCL1B	−4.70516	−2.58147	−2.2034
7003	TEA DOMAIN FAMILY MEMBER 1 (SV40 TRANSCRIPTIONAL ENHANCER	TEAD1	−3.55384	−2.12333
	FACTOR)
7006	TEC PROTEIN TYROSINE KINASE	TEC	−1.85863		−1.90836
23371	TENSIN LIKE C1 DOMAIN CONTAINING PHOSPHATASE (TENSIN 2)	TENC1	−3.23649	−2.03696
7011	TELOMERASE-ASSOCIATED PROTEIN 1	TEP1	−2.33241	−5.04989
7022	TRANSCRIPTION FACTOR AP-2 GAMMA (ACTIVATING ENHANCER	TFAP2C		−2.50319	−2.92926
	BINDING PROTEIN 2 GAMMA)
29842	TRANSCRIPTION FACTOR CP2-LIKE 1	TFCP2L1	−2.75048	−2.80252
29844	TCF3 (E2A) FUSION PARTNER (IN CHILDHOOD LEUKEMIA)	TFPT	−3.11717	−3.06358	−3.10235
51497	TH1-LIKE (DROSOPHILA)	TH1L	−2.06756	−2.99752
353376	TOLL-LIKE RECEPTOR ADAPTOR MOLECULE 2	TICAM2	−2.54594	−1.92781
7082	TIGHT JUNCTION PROTEIN 1 (ZONA OCCLUDENS 1)	TJP1	−1.99226		−3.30745
7083	THYMIDINE KINASE 1, SOLUBLE	TK1	−2.50739	−2.82968
9874	TOUSLED-LIKE KINASE 1	TLK1		−2.30245	−2.34146
7092	TOLLOID-LIKE 1	TLL1	−2.42975	−2.34648
51284	TOLL-LIKE RECEPTOR 7	TLR7	−3.20969	−2.41105
9032	TRANSMEMBRANE 4 L SIX FAMILY MEMBER 5	TM4SF5		−2.0935	−2.68228
53346	TRANSMEMBRANE 6 SUPERFAMILY MEMBER 1	TM6SF1	−4.00854	−2.48158
51643	TRANSMEMBRANE BAX INHIBITOR MOTIF CONTAINING 4	TMBIM4	−3.44325	−1.93345
79905	TRANSMEMBRANE CHANNEL-LIKE 7	TMC7	−2.27616	−1.85573	−1.88393
55002	TRANSMEMBRANE AND COILED-COIL DOMAINS 3	TMCO3	−2.37384	−1.86547	−2.07757
8834	TRANSMEMBRANE PROTEIN 11	TMEM11	−4.15914	−4.43887
144404	HYPOTHETICAL LOC144404	TMEM120B	−5.92348	−2.20834	−2.36457
85014	HYPOTHETICAL PROTEIN MGC14141	TMEM141	−3.93132	−2.00305
51522	TRANSMEMBRANE PROTEIN 14C	TMEM14C	−3.83709	−3.00587
201799	HYPOTHETICAL PROTEIN FLJ32028	TMEM154	−2.9453	−4.0793	−3.83264
55858	TPA REGULATED LOCUS	TMEM165	−3.00582	−2.11807
84286	HYPOTHETICAL PROTEIN MGC4618	TMEM175		−2.30215	−1.95875
84548	FAMILY WITH SEQUENCE SIMILARITY 11, MEMBER A	TMEM185A	−2.878	−1.92037
387521	UBIQUITIN-CONJUGATING ENZYME VARIANT KUA	TMEM189	−2.38212	−3.47634
55161	TRANSMEMBRANE PROTEIN 33	TMEM33		−1.85009	−3.18274
131616	TRANSMEMBRANE PROTEIN 42	TMEM42	−4.81996	−2.71992
51249	TRANSMEMBRANE PROTEIN 69	TMEM69	−1.86253	−2.34459
144110	TRANSMEMBRANE PROTEIN 86A	TMEM86A	−3.98965	−2.15319
641649	TRANSMEMBRANE PROTEIN 91	TMEM91	−2.39084		−3.8996
126259	HYPOTHETICAL PROTEIN MGC23244	TMIGD2	−2.19155	−3.54299
29766	TROPOMODULIN 3 (UBIQUITOUS)	TMOD3	−2.90096	−3.05522
344805	TRANSMEMBRANE PROTEASE, SERINE 7	TMPRSS7	−2.22643	−2.42787
160335	TRANSMEMBRANE AND TETRATRICOPEPTIDE REPEAT CONTAINING 2	TMTC2	−3.26585		−2.1169
8600	TUMOR NECROSIS FACTOR (LIGAND) SUPERFAMILY, MEMBER 11	TNFSF11	−2.5807	−5.88377
23534	TRANSPORTIN 3	TNPO3	−4.90495	−4.06996	−3.75153
10140	TRANSDUCER OF ERBB2, 1	TOB1	−2.4024		−2.0188
9804	TRANSLOCASE OF OUTER MITOCHONDRIAL MEMBRANE 20 HOMOLOG	TOMM20		−3.38974	−2.88057
	(YEAST)
56993	TRANSLOCASE OF OUTER MITOCHONDRIAL MEMBRANE 22 HOMOLOG	TOMM22		−2.1434	−2.49763
	(YEAST)
58476	TUMOR PROTEIN P53 INDUCIBLE NUCLEAR PROTEIN 2	TP53INP2	−2.66254	−4.08177	−2.05149
53373	TWO PORE SEGMENT CHANNEL 1	TPCN1	−2.18053	−2.14121	−2.27751
7164	TUMOR PROTEIN D52-LIKE 1	TPD52L1	−3.5544	−3.25758
7173	THYROID PEROXIDASE	TPO	−2.12718	−2.65698
51693	HEMATOPOIETIC STEM/PROGENITOR CELLS 176	TRAPPC2L	−2.35046	−2.84987
54210	TRIGGERING RECEPTOR EXPRESSED ON MYELOID CELLS 1	TREM1	−2.41212	−4.62915	−2.14621
55809	TRANSCRIPTIONAL REGULATING FACTOR 1	TRERF1	−2.90946	−4.16991
8805	TRIPARTITE MOTIF-CONTAINING 24	TRIM24	−1.97462	−1.93153
10155	TRIPARTITE MOTIF-CONTAINING 28	TRIM28		−2.13008	−1.92375
166655	TRIPARTITE MOTIF-CONTAINING 60	TRIM60	−1.98769	−2.65534
55128	TRIPARTITE MOTIF-CONTAINING 68	TRIM68	−2.27145	−3.97537
6738	TROVE DOMAIN FAMILY, MEMBER 2	TROVE2	−2.34569		−2.09889
7222	TRANSIENT RECEPTOR POTENTIAL CATION CHANNEL, SUBFAMILY C,	TRPC3	−2.25341		−2.81435
	MEMBER 3
7225	TRANSIENT RECEPTOR POTENTIAL CATION CHANNEL, SUBFAMILY C,	TRPC6	−2.47758	−2.44406	−2.91983
	MEMBER 6
140803	HYPOTHETICAL PROTEIN FLJ20087	TRPM6	−3.44552	−1.88594	−2.78798
57616	ZINC FINGER PROTEIN 537	TSHZ3	−1.90786	−3.21475	−2.10168
7259	TSPY-LIKE 1	TSPYL1	−2.56945	−3.16215	−2.20417
7264	TISSUE SPECIFIC TRANSPLANTATION ANTIGEN P35B	TSTA3	−4.00686	−2.37017
26140	TUBULIN TYROSINE LIGASE-LIKE FAMILY, MEMBER 3	TTLL3	−1.90887	−1.88575
84617	TUBULIN, BETA 6	TUBB6	−2.01658	−2.87146
7284	TU TRANSLATION ELONGATION FACTOR, MITOCHONDRIAL	TUFM	−2.92069	−2.19069
7294	TXK TYROSINE KINASE	TXK	−2.78577	−3.11976
200081	TAXILIN ALPHA	TXLNA		−2.47492	−2.55174
54957	THIOREDOXIN-LIKE 4B	TXNL4B	−1.96591	−2.3762	−3.21574
7326	UBIQUITIN-CONJUGATING ENZYME E2G 1 (UBC7 HOMOLOG, YEAST)	UBE2G1	−1.86023	−2.46231	−2.00714
7326	UBIQUITIN-CONJUGATING ENZYME E2G 1 (UBC7 HOMOLOG, YEAST)	UBE2G1	−1.86023	−2.46231	−2.00714
9246	UBIQUITIN-CONJUGATING ENZYME E2L 6	UBE2L6	−4.53705	−3.29012
143630	HYPOTHETICAL PROTEIN MGC20470	UBQLNL		−1.96977	−3.45196
92181	DENDRITIC CELL-DERIVED UBIQUITIN-LIKE PROTEIN	UBTD2		−3.05707	−1.8902
7993	UBX DOMAIN CONTAINING 6	UBXD6	−2.07223	−2.28693
51569	UBIQUITIN-FOLD MODIFIER 1	UFM1		−2.44738	−2.89156
167127	UDP GLYCOSYLTRANSFERASE 3 FAMILY, POLYPEPTIDE A2	UGT3A2	−2.29205	−1.92947
121665	DKFZP586C1324 PROTEIN	UNQ1887		−2.58128	−2.02877
7398	UBIQUITIN SPECIFIC PEPTIDASE 1	USP1	−1.91599	−2.7275	−2.81914
11274	UBIQUITIN SPECIFIC PEPTIDASE 18	USP18	−3.79525	−3.50393	−3.57991
9960	UBIQUITIN SPECIFIC PEPTIDASE 3	USP3	−3.70597		−2.06201
9098	HYPERPOLYMORPHIC GENE 1	USP6	−1.85927	−1.99963
7405	UV RADIATION RESISTANCE ASSOCIATED GENE	UVRAG	−3.54039	−4.113
8673	VESICLE-ASSOCIATED MEMBRANE PROTEIN 8 (ENDOBREVIN)	VAMP8	−1.86941		−2.06279
50853	VILLIN-LIKE	VILL	−2.94995		−1.92166
7433	VASOACTIVE INTESTINAL PEPTIDE RECEPTOR 1	VIPR1	−5.80786	−4.69544	−3.96241
79720	VACUOLAR PROTEIN SORTING 37B (YEAST)	VPS37B	−1.85816	−2.02574
55275	VACUOLAR PROTEIN SORTING 53 (YEAST)	VPS53	−3.0736		−3.25681
128434	CHROMOSOME 20 OPEN READING FRAME 102	VSTM2L	−2.56128		−2.36445
340706	VON WILLEBRAND FACTOR A DOMAIN CONTAINING 2	VWA2	−2.6436	−1.88033
11193	WW DOMAIN BINDING PROTEIN 4 (FORMIN BINDING PROTEIN 21)	WBP4	−2.88433		−1.89074
55759	WD REPEAT DOMAIN 12	WDR12	−2.06357	−2.34823
79269	WD REPEAT DOMAIN 32	WDR32	−2.20716	−2.26755
55339	WD REPEAT DOMAIN 33	WDR33	−2.97558		−1.90321
401551	SIMILAR TO HYPOTHETICAL PROTEIN FLJ25955	WDR38	−1.87216	−2.76576	−3.11598
139170	WD REPEAT DOMAIN 40B	WDR40B	−2.30236		−2.23209
50717	WD REPEAT DOMAIN 42A	WDR42A	−2.37447	−2.61814	−2.94676
348793	WD REPEAT DOMAIN 53	WDR53	−4.71096	−4.37789
55100	WD REPEAT DOMAIN 70	WDR70	−2.00896		−2.72161
79819	WD REPEAT DOMAIN 78	WDR78	−3.82026	−4.04859
23038	WD AND TETRATRICOPEPTIDE REPEATS 1	WDTC1	−2.08283	−2.63992	−2.30554
147179	WIRE PROTEIN	WIPF2	−2.63385	−2.09502
80014	WW, C2 AND COILED-COIL DOMAIN CONTAINING 2	WWC2	−5.43289		−2.11183
51741	PUTATIVE OXIDOREDUCTASE	WWOX	−2.98849		−2.36351
2829	CHEMOKINE (C MOTIF) RECEPTOR 1	XCR1	−2.09586	−2.71649
286046	CHROMOSOME 8 OPEN READING FRAME 7	XKR6	−2.09863	−1.89435
91419	XRCC6 BINDING PROTEIN 1	XRCC6BP1	−2.27418	−1.92721
541465	CANCER/TESTIS ANTIGEN CT45-2	XX-	−2.78451	−2.59265	−2.14218
		FW88277B6.1
10652	SNARE PROTEIN YKT6	YKT6	−2.29434		−2.30459
79693	YRDC DOMAIN CONTAINING (E. COLI)	YRDC	−3.01605	−2.64556
253943	YTH DOMAIN FAMILY, MEMBER 3	YTHDF3	−2.76914	−2.84077
7532	TYROSINE 3-MONOOXYGENASE/TRYPTOPHAN 5-MONOOXYGENASE	YWHAG	−3.31557	−2.31734
	ACTIVATION PROTEIN, GAMMA POLYPEPTIDE
7528	YY1 TRANSCRIPTION FACTOR	YY1	−3.29105	−3.64289
57684	ZINC FINGER AND BTB DOMAIN CONTAINING 26	ZBTB26	−3.34289	−3.49875
9877	ZINC FINGER CCCH-TYPE CONTAINING 11A	ZC3H11A	−2.84078		−3.43356
84240	ZINC FINGER, CCHC DOMAIN CONTAINING 9	ZCCHC9	−3.2696	−2.5597
84936	ZINC FINGER, FYVE DOMAIN CONTAINING 19	ZFYVE19	−2.05237	−1.99609
84217	ZINC FINGER, MYND-TYPE CONTAINING 12	ZMYND12	−2.94854	−3.47689	−2.51531
118490	ZINC FINGER, MYND-TYPE CONTAINING 17	ZMYND17		−2.47856	−1.89741
7690	ZINC FINGER PROTEIN 131 (CLONE PHZ-10)	ZNF131	−1.88418	−2.64243	−2.1069
7766	ZINC FINGER PROTEIN 223	ZNF223	−1.97433	−2.11869
7572	ZINC FINGER PROTEIN 24 (KOX 17)	ZNF24	−2.06908	−2.42245
10224	ZINC FINGER PROTEIN 443	ZNF443	−2.11094	−2.36031
114821	ZINC FINGER PROTEIN 452	ZNF452	−3.09253	−3.53396	−2.54663
284443	ZINC FINGER PROTEIN 493	ZNF493	−2.27517		−2.61513
22869	ZINC FINGER PROTEIN 510	ZNF510	−5.67033	−2.21998	−2.82731
25925	ZINC FINGER PROTEIN 521	ZNF521	−2.85699	−2.52397
147741	ZINC FINGER PROTEIN 560	ZNF560	−2.22818	−2.81243	−2.94421
284346	ZINC FINGER PROTEIN 575	ZNF575	−2.32433	−2.1127	−2.03963
169270	ZINC FINGER PROTEIN 596	ZNF596		−2.20356	−2.17493
121274	ZINC FINGER PROTEIN 641	ZNF641	−1.89321	−2.1958
146542	ZINC FINGER PROTEIN 688	ZNF688		−3.74763	−2.11944
163051	ZINC FINGER PROTEIN 709	ZNF709	−2.23282		−1.86393
7627	ZINC FINGER PROTEIN 75A	ZNF75A	−3.62969	−4.07762
7629	ZINC FINGER PROTEIN 76 (EXPRESSED IN TESTIS)	ZNF76	−2.05861	−2.09435
7633	ZINC FINGER PROTEIN 79 (PT7)	ZNF79	−3.36041	−2.94994
30834	ZINC RIBBON DOMAIN CONTAINING, 1	ZNRD1	−2.01992	−3.05193
23140	ZINC FINGER, ZZ-TYPE WITH EF-HAND DOMAIN 1	ZZEF1		−1.90416	−2.10696

One or more currently preferred embodiments have been described by way of example. It will be apparent to persons skilled in the art that a number of variations and modifications can be made without departing from the scope of the invention as defined in the claims.

Claims

1. A method of reducing viability of a tumor cell in a subject, comprising the steps of: introducing into a tumor cell in said subject an agent effective to modulate endoplasmic reticulum (ER) stress response and sensitize the tumour cell to cytolytic activity of an oncolytic virus in said subject; andcontacting the tumor cell with an oncolytic virus in an amount effective to reduce viability of the sensitized tumour cell,wherein viability of the tumor cell is reduced.

2. The method of claim 1, wherein the agent is effective to enhance, diminish or inhibit the ER stress response in said subject.

3. (canceled)

4. The method of claim 1, wherein the agent is a siRNA specific to ERN, ATF6, Derlin1, Derlin2 or SEC61, a molecule effective to modulate ERN, ATF6, Derlin1, Derlin2 or SEC61 signaling, or a modified oncolytic virus wherein said modification renders the oncolytic virus effective to modulate ER stress response and sensitize the tumour cell to cytolytic activity.

5. (canceled)

6. The method of claim 1, wherein the tumour cell or cancer cell is selected from colon cancer, lung cancer, liver cancer, prostate cancer, bladder cancer, neck and mouth cancer, breast cancer, glioblastoma, lymphoma, carcinoma, renal cell cancer, pancreatic cancer, and ovarian cancer cells.

7. The method of claim 1, wherein the oncolytic virus is a native or modified herpes virus, Adenovirus, Adeno-associated virus, influenza virus, reovirus, rhabdovirus, Newcastle virus, vaccinia virus, poliovirus, measles virus, mumps virus, sindbis virus (SIN) or sendai virus (SV).

8. The method of claim 7, wherein the oncolytic virus is a native or modified rhabdovirus.

9. The method of claim 8, wherein the oncolytic virus is a native or modified vesicular stomatitis virus (VSV) or Maraba virus.

10. The method of claim 9, wherein the virus is a mutant virus modified with a function-improving mutation to make the virus a more effective cancer or tumour cell lysing agent.

11. A method of modulating sensitivity of cancer cells to infection by an oncolytic virus, the method comprising introducing into a cancer cell an agent effective to modulate endoplasmic reticulum (ER) stress response and sensitize the cancer cell to cytolytic activity of the oncolytic virus, wherein the cancer cells are sensitized to infection by the oncolytic virus.

12. A method of identifying a tumour cell sensitizing agent effective for sensitizing tumour cells to infection by an oncolytic virus, comprising: providing a test molecule with putative endoplasmic reticulum (ER) stress response modulating activity,adding the test molecule to a sample of said tumor cells,contacting the tumor cells with the oncolytic virus, andcomparing cytolytic activity of the oncolytic virus in the sample of tumour cells with the test molecule to activity of the oncolytic virus in a sample of tumour cells without the test molecule,wherein increased cytolytic activity of the oncolytic virus in the sample of tumour cells with the test molecule indicates the presence of a tumour cell sensitizing agent.

13. A compound effective to modulate endoplasmic reticulum (ER) stress response and sensitize a tumour cell to cytolytic activity of an oncolytic virus in a subject.

14. The compound of claim 13, wherein the compound is effective to inhibit the ER stress response in said subject.

15. The compound of claim 13, wherein the compound is a siRNA specific to ERN, ATF6, Derlin1, Derlin2 or SEC61 or a molecule effective to modulate ERN, ATF6, Derlin1, Derlin2 or SEC61 signaling.

16. The compound of claim 15, wherein the compound is effective to block or enhance ERN, ATF6, Derlin1, Derlin2 or SEC61 signaling.

17. The compound of claim 16, wherein the compound is a protein, a small molecule, a nucleic acid, or an antibody.

18. A composition comprising the compound of claim 13 and an acceptable carrier or excipient.

19. (canceled)

20. (canceled)

21. (canceled)

22. (canceled)

23. (canceled)

24. (canceled)

25. (canceled)

26. (canceled)

27. A method of sensitizing a tumor to cytolytic activity of an oncolytic virus, said method comprising inducing in a subject a mild stress to the endoplasmic reticulum (ER).

28. The method of claim 27, wherein inducing the mild stress comprises genetically disrupting an ER stress response gene.

29. The method of claim 28, wherein the ER stress response gene is selected from the group consisting of IRE1/ERN, DERLIN, and ATF6.

30. The method of claim 27, wherein inducing the mild stress comprises chemically inhibiting IRE1/ERN1

31. The method of claim 30, wherein compound 2 is administered to the subject to chemically inhibit IRE1/ERN1.

32. (canceled)

33. (canceled)

34. (canceled)

35. (canceled)

36. (canceled)

37. (canceled)

38. (canceled)

39. (canceled)