HEPATITIS C VIRUS INFECTION BIOMARKERS

TECHNICAL FIELD

This invention relates to hepatitis C virus (HCV) infection, and more particularly to a signature set of HCV infection.

BACKGROUND

Infection by hepatitis C virus (“HCV”) is a compelling human medical problem. HCV is recognized as the causative agent for most cases of non-A, non-B hepatitis, with an estimated human sero-prevalence of 3% globally (A. Alberti et al., “Natural History of Hepatitis C,” (1999) J. Hepatology, 31, (Suppl. 1), pp. 17-24). Nearly four million individuals may be infected in the United States alone (M. J. Alter et al., “The Epidemiology of Viral Hepatitis in the United States,” (1994) Gastroenterol. Clin. North Am., 23, pp. 437-455; M. J. Alter “Hepatitis C Virus Infection in the United States,” (1999) J. Hepatology, 31, (Suppl. 1), pp. 88-91).

Upon first exposure to HCV only about 20% of infected individuals develop acute clinical hepatitis while others appear to resolve the infection spontaneously. In almost 70% of instances, however, the virus establishes a chronic infection that persists for decades (S. Iwarson, “The Natural Course of Chronic Hepatitis,” (1994) FEMS Microbiology Reviews, 14, pp. 201-204; D. Lavanchy, “Global Surveillance and Control of Hepatitis C,” (1999) J. Viral Hepatitis, 6, pp. 35-47). This usually results in recurrent and progressively worsening liver inflammation, which often leads to more severe disease states such as cirrhosis and hepatocellular carcinoma (M. C. Kew, “Hepatitis C and Hepatocellular Carcinoma”, (1994) FEMS Microbiology Reviews, 14, pp. 211-220; I. Saito et. al., “Hepatitis C Virus Infection is Associated with the Development of Hepatocellular Carcinoma,” (1990) Proc. Natl. Acad. Sci. USA, 87, pp. 6547-6549). It is estimated that HCV infects 170 million persons worldwide. Over the next ten years, as a larger proportion of patients who are currently infected enter the third decade of their infection, the number of deaths attributed to hepatitis C is expected to significantly increase. Unfortunately, there are no broadly effective treatments for the debilitating progression of chronic HCV.

SUMMARY

The inventors have identified a set of genes, e.g., a signature set, associated with HCV infection. The inventors have also determined that the anti-viral activity of VX-950 results in changes in gene expression, e.g., treatment with VX-950 leads to normalization of the signature set such that the gene transcript levels after 14 days of treatment more closely resemble levels seen in non-infected subjects. Further, the inventors have established a baseline gene expression set which includes genes, e.g., interferon-sensitive genes (ISGs) that can be monitored and correlated with (and optionally, predictive of) treatment, e.g., VX-950 dosing, outcomes.

In one aspect, the disclosure features a method of evaluating a subject (e.g., a subject suspected of having a viral infection, e.g., HCV infection), e.g., for the presence or level of hepatitis C virus (HCV) infection (e.g., chronic HCV). The method includes providing an evaluation of the expression of the genes in a signature set of genes in the subject, wherein the signature set has the following properties: it includes a plurality of genes each of which is differentially expressed as between virally infected individuals and non-infected individuals and it contains a sufficient number of differentially expressed genes such that differential expression (e.g., as compared to a non-infected reference) of each of the genes in the signature set in a subject is predictive of infection with no more than about 15, about 10, about 5, about 2.5, or about 1% false positives (wherein false positive means identifying a subject as virus infected when the subject is not infected); and providing a comparison of the expression of each of the genes in the set from the subject with a reference value, thereby evaluating the subject.

In some embodiments, the comparison includes comparing expression in the subject with a non-infected reference and wherein differential expression of each of the genes in the signature set of genes indicates, a first state, e.g., infection or a first likelihood of infection, and differential expression of less than all of the genes in the signature set indicates a second state, e.g., non-infection or a second likelihood of infection.

In some embodiments, the reference is a value of expression from one or more, e.g., a cohort of, uninfected subjects.

In some embodiments, the comparison includes comparing the expression in the subject with an infected reference and wherein non-differential (e.g., similar) expression of each of the genes in the signature set of genes indicates a first state, e.g., infection or a first likelihood of infection, and non-differential (e.g., similar) expression of less than all of the genes in the signature set indicates a second state, e.g., non-infection or a second likelihood of infection.

In some embodiments, the reference is a value of expression from one or more, e.g., a cohort of, virally infected subjects.

In some embodiments, peripheral blood from the subject is evaluated.

In some embodiments, the evaluating occurs prior to administering an inhibitor of a viral protease to the subject.

In other embodiments, the evaluating occurs during the course of administering or after administering an inhibitor of a viral protease to the subject (optionally in combination with evaluating prior to administering the inhibitor).

In some embodiments, the inhibitor is VX-950, SCH-503034, or BILN-261 (ciluprevir).

In some embodiments, the method includes determining a post administration level of gene expression, determined, e.g., at the RNA or protein level, for an interferon sensitive gene (ISG) in the subject to provide a post administration determined value; and comparing the post administration determined value with a reference value, (by way of example, the reference value can be the level of expression of the ISG prior to administration of the antiviral treatment), thereby evaluating the subject, e.g., determining if the subject is an enhanced responder or a non-enhanced responder.

In some embodiments, the method includes determining a pre administration level of gene expression, determined, e.g., at the RNA or protein level, for an interferon sensitive gene (ISG) in the subject to provide a pre administration determined value; and comparing the pre administration determined value with a reference value, (by way of example, the reference value can be the level of expression of the ISG after commencing administration of the antiviral treatment), thereby evaluating the subject, e.g., determining if the subject is an enhanced responder or a non-enhanced responder.

In some embodiments, the signature set of genes includes a plurality of genes associated with hepatitis C virus (HCV) infection (e.g., chronic infection). In some embodiments, the signature set of genes includes a plurality of genes listed in Table 2. In some embodiments, the signature set of genes includes at least about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, about 95%, about 96%, about 97%, about 98, or about 99% of the genes listed in Table 2.

In some embodiments, the signature set of genes includes a gene from one or more, e.g., each of the following categories (e.g., ontology categories): organismal physiological processes; immune response (e.g., IFIT2, IFIT3, IFIT4, IFI5, IFT16, IFT27, IFT30, IFT35, IFT44, IFITM1, IFITM2, IFITM3, MX1); defense response (e.g., ITGB1); response to biotic stimulus (e.g., CCR1); response to stimulus (e.g., OGG1); response to stress (e.g., CEBP/B); response to pest, pathogen, or parasite (e.g., IFT27); or response to virus (e.g., IRF7, PLSCR1). In some embodiments, the signature set of genes includes a gene from each of 2, 3, 4, 5, 6, 7, or 8 gene ontology categories described herein. In some embodiments, the signature set of genes includes a plurality of genes from each of 2, 3, 4, 5, 6, 7, or 8 gene ontology categories described herein.

In some embodiments, the signature set of genes includes one or more interferon-sensitive genes (ISG). In some embodiments, the ISG is selected from the group consisting of: IFIT1, RSAD2, IFIT2, IFI16, IFI44, IFIT2, IFIT5, PLSCR1, IFIT3, IFI35, IFITM1, IFITM3, IFI30, IFITM1, IFITM2, GIP2, OAS3, IFIT3, MX1, IFIL44L, IFI27, IFIT2A, PRSAD, or IFITA. In some embodiments, the signature set of genes includes at least 1, 2, 3, 4, 5, 6, 7, 8, 9, or all of: GIP2, OAS3, IFIT3, MX1, IFIL44L, PLSCR1, IFI27, IFIT2A, PRSAD, or IFITA.

In some embodiments, the signature set of genes includes at least 20, 40, 60, 80, 100, 150, or 200 genes.

In other embodiments, the signature set of genes includes no more than 20, 40, 60, 80, 100, 150, or 200 genes.

In some embodiments, the signature set of genes includes the genes listed in Table 2.

In some embodiments, the signature set of genes includes at least 10, 20, 30, 40, or 50 genes which are more highly expressed in infection than in non infection.

In other embodiments, the signature set of genes includes at least 10, 20, 30, 40, or 50 genes which are more highly expressed in non-infection than in infection.

In some embodiments, the method includes assigning the subject to a diagnostic class.

In some embodiments, the method includes selecting the subject for a treatment.

In some embodiments, the method further includes providing the evaluation to the subject, a third party payer, an insurance company, employer, employer sponsored health plan, HMO, governmental entity, healthcare provider, a treating physician, an HMO, a hospital, an entity which sells or supplies a drug.

In one aspect, the disclosure features a method of evaluating the efficacy of a treatment of HCV infection (e.g., chronic HCV) in a subject. The method includes administering the treatment; and performing an evaluation described herein, thereby evaluating the efficacy of the treatment.

In some embodiments, the method includes providing a determination of a first level of gene expression associated with HCV infection in the subject at a first time point (e.g., wherein the first time point is prior to, or within about 1, 2, 3, 4, or 5 days of the commencement of, administration of an anti-HCV therapy (e.g., an HCV protease inhibitor, e.g., VX-950)); providing a determination of a second level of gene expression in the subject at a second time point after the first time point and preferably the second time point is after commencement of administration of anti-HCV therapy (e.g., wherein the second time point is taken at least 1, 2, 3, 4, 5, or more days after the first time point or wherein the second time point is 7, 8, 9, 10, 11, 12, 13, 14 or more days after the commencement of administration of the anti-HCV therapy); and providing a comparison of the first and second levels of gene expression, wherein sustained levels of gene expression (e.g., the levels differ by no more than about 60%, about 50%, about 40%, about 30%, about 20%, about 10%, about 5%, about 2%, or about 1%) between the first and second time points is indicative of effective treatment.

In some embodiments, providing a comparison of the first and second levels of gene expression includes a comparison of the levels of one or more interferon-sensitive genes (ISG). In some embodiments, the ISG is selected from the group consisting of: IFIT1, RSAD2, IFIT2, IFT16, IFT44, IFIT2, IFIT5, PLSCR1, IFIT3, IFT35, IFITM1, IFITM3, IFT30, IFITM1, IFITM2, GIP2, OAS3, IFIT3, MX1, IFIL44L, IFI27, IFIT2A, PRSAD, or IFITA. In some preferred embodiments, first and second levels of at least 1, 2, 3, 4, 5, 6, 7, 8, 9, or all of: GIP2, OAS3, IFIT3, MX1, IFIL44L, PLSCR1, IFI27, IFIT2A, PRSAD, or IFITA are compared.

In another aspect, the disclosure features a method of evaluating the efficacy of a treatment of HCV infection (e.g., chronic HCV) in a subject. The method includes providing a determination of a first level of gene expression associated with HCV infection in the subject at a first time point (e.g., wherein the first time point is prior to, or within about 1, 2, 3, 4, or 5 days of the commencement of, administration of an anti-HCV therapy (e.g., an HCV protease inhibitor, e.g., VX-950)); providing a determination of a second level of gene expression in the subject at a second time point after the first time point and preferably the second time point is after commencement of administration of anti-HCV therapy (e.g., wherein the second time point is taken at least 1, 2, 3, 4, 5, or more days after the first time point or wherein the second time point is 7, 8, 9, 10, 11, 12, 13, 14 or more days after the commencement of administration of the anti-HCV therapy); and providing a comparison of the first and second levels of gene expression to a control level of gene expression, wherein a smaller difference between the second level and the control level as compared to the difference between the first level and the control level is indicative of effective treatment.

In some embodiments, the control corresponds to the level in a non-HCV infected subject or in a cohort of non-infected subjects.

In another aspect, the disclosure features a method of evaluating the efficacy of a drug for use in treatment of HCV infection (e.g., chronic HCV) in a subject. The method includes providing a determination of a first level of gene expression associated with HCV infection in the subject at a first time point (e.g., wherein the first time point is prior to, or within about 1, 2, 3, 4, or 5 days of the commencement of, administration of an anti-HCV therapy (e.g., an HCV protease inhibitor, e.g., VX-950)); providing a determination of a second level of gene expression in the subject at a second time point after the first time point and preferably the second time point is after commencement of administration of anti-HCV therapy (e.g., wherein the second time point is taken at least 1, 2, 3, 4, 5, or more days after the first time point or wherein the second time point is 7, 8, 9, 10, 11, 12, 13, 14 or more days after the commencement of administration of the anti-HCV therapy); and providing a comparison of the first and second levels of gene expression, wherein sustained levels of gene expression (e.g., the levels differ by no more than about 60%, about 50%, about 40%, about 30%, about 20%, about 10%, about 5%, about 2%, or about 1%) between the first and second time points is indicative of drug efficacy.

In some embodiments, the comparison of the first and second levels of gene expression includes comparing the levels of one or more interferon-sensitive genes (ISG). In some embodiments, the ISG is selected from the group consisting of: IFIT1, RSAD2, IFIT2, IFT16, IFT44, IFIT2, IFIT5, PLSCR1, IFIT3, IFT35, IFITM1, IFITM3, IFI30, IFITM1, IFITM2, GIP2, OAS3, IFIT3, MX1, IFIL44L, IFI27, IFIT2A, PRSAD, or IFITA. In some preferred embodiments, first and second levels of at least 1, 2, 3, 4, 5, 6, 7, 8, 9, or all of: GIP2, OAS3, IFIT3, MX1, IFIL44L, PLSCR1, IFI27, IFIT2A, PRSAD, or IFITA are compared.

In another aspect, the disclosure features a method of evaluating the efficacy of a drug for use in treatment of HCV infection (e.g., chronic HCV) in a subject. The method includes providing a determination of a first level of gene expression associated with HCV infection in the subject at a first time point (e.g., wherein the first time point is prior to, or within about 1, 2, 3, 4, or 5 days of the commencement of, administration of an anti-HCV therapy (e.g., an HCV protease inhibitor, e.g., VX-950)); providing a determination of a second level of gene expression in the subject at a second time point after the first time point and preferably the second time point is after commencement of administration of anti-HCV therapy (e.g., wherein the second time point is taken at least 1, 2, 3, 4, 5, or more days after the first time point or wherein the second time point is 7, 8, 9, 10, 11, 12, 13, 14 or more days after the commencement of administration of the anti-HCV therapy); and providing a comparison of the first and second levels of gene expression to a control level of gene expression, wherein a smaller difference between the second level and the control level as compared to the difference between the first level and the control level is indicative of drug efficacy.

In some embodiments, the gene expression associated with HCV infection is determined for a plurality of the genes listed in Table 2.

In some embodiments, the plurality includes at least about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, about 95%, about 96%, about 97%, about 98, or about 99% of the genes listed in Table 2. In some embodiments, the plurality includes the genes listed in Table 2.

In some embodiments, the plurality includes a gene from one or more, e.g., each of the following categories (e.g., ontology categories): organismal physiological processes; immune response (e.g., IFIT2, IFIT3, IFIT4, IFI5, IFT16, IFT27, IFT30, IFT35, IFT44, IFITM1, IFITM2, IFITM3, MX1); defense response (e.g., ITGB1); response to biotic stimulus (e.g., CCR1); response to stimulus (e.g., OGG1); response to stress (e.g., CEBP/B); response to pest, pathogen, or parasite (e.g., IFT27); or response to virus (e.g., IRF7, PLSCR1). In some embodiments, the plurality includes a gene from each of 2, 3, 4, 5, 6, 7, or 8 gene ontology categories described herein. In some embodiments, the plurality includes a plurality of genes from each of 2, 3, 4, 5, 6, 7, or 8 gene ontology categories described herein.

In another aspect, the disclosure features a method of monitoring treatment for HCV infection (e.g., chronic HCV) in a subject and includes administering the treatment (e.g., a treatment described herein), performing an evaluation described herein, thereby monitoring the treatment.

In some embodiments, the method includes providing a determination of a first level of gene expression associated with HCV infection in the subject at a first time point (e.g., wherein the first time point is prior to, or within about 1, 2, 3, 4, or 5 days of the commencement of, administration of an anti-HCV therapy (e.g., an HCV protease inhibitor, e.g., VX-950)); providing a determination of a second level of gene expression in the subject at a second time point after the first time point and preferably the second time point is after commencement of administration of anti-HCV therapy (e.g., wherein the second time point is taken at least 1, 2, 3, 4, 5, or more days after the first time point or wherein the second time point is 7, 8, 9, 10, 11, 12, 13, 14 or more days after the commencement of administration of the anti-HCV therapy); providing a comparison of the first and second levels of gene expression; and providing a determination of whether levels of gene expression are sustained (e.g., the levels differ by no more than about 60%, about 50%, about 40%, about 30%, about 20%, about 10%, about 5%, about 2%, or about 1%) between the first and second time points, thereby monitoring the treatment.

In some embodiments, the comparison of the first and second levels of gene expression includes comparing the levels of one or more interferon-sensitive genes (ISG). In some embodiments, the ISG is selected from the group consisting of: IFIT1, RSAD2, IFIT2, IFT16, IFT44, IFIT2, IFIT5, PLSCR1, IFIT3, IFT35, IFITM1, IFITM3, IFT30, IFITM1, IFITM2, GIP2, OAS3, IFIT3, MX1, IFIL44L, IFI27, IFIT2A, PRSAD, or IFITA. In some preferred embodiments, first and second levels of at least 1, 2, 3, 4, 5, 6, 7, 8, 9, or all of: GIP2, OAS3, IFIT3, MX1, IFIL44L, PLSCR1, IFI27, IFIT2A, PRSAD, or IFITA are compared.

In another aspect, the disclosure features a method of monitoring treatment for HCV infection (e.g., chronic HCV) in a subject. The method includes providing a determination of a first level of gene expression associated with HCV infection in the subject at a first time point (e.g., wherein the first time point is prior to, or within about 1, 2, 3, 4, or 5 days of the commencement of, administration of an anti-HCV therapy (e.g., an HCV protease inhibitor, e.g., VX-950)); providing a determination of a second level of gene expression in the subject at a second time point after the first time point and preferably the second time point is after commencement of administration of anti-HCV therapy (e.g., wherein the second time point is taken at least 1, 2, 3, 4, 5, or more days after the first time point or wherein the second time point is 7, 8, 9, 10, 11, 12, 13, 14 or more days after the commencement of administration of the anti-HCV therapy); and providing a comparison of the first and second levels of gene expression to a control level of the gene transcript, thereby monitoring the treatment.

In some embodiments, the gene expression associated with HCV infection is determined for a plurality of the genes listed in Table 2. In some embodiments, the plurality includes at least about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, about 95%, about 96%, about 97%, about 98, or about 99% of the genes listed in Table 2. In some embodiments, the plurality includes the genes listed in Table 2.

In some embodiments, the plurality comprises a gene from each of 2, 3, 4, 5, 6, 7, or 8 gene ontology categories described herein.

In one aspect, the disclosure features a method of evaluating a drug candidate for treatment of HCV infection (e.g., chronic HCV) in a subject. The method includes providing a determination of a first level of gene expression associated with HCV infection in the subject at a first time point (e.g., wherein the first time point is prior to, or within about 1, 2, 3, 4, or 5 days of the commencement of, administration of an anti-HCV therapy (e.g., an HCV protease inhibitor, e.g., VX-950)); providing a determination of a second level of gene expression in the subject at a second time point after the first time point and preferably the second time point is after commencement of administration of anti-HCV therapy (e.g., wherein the second time point is taken at least 1, 2, 3, 4, 5, or more days after the first time point or wherein the second time point is 7, 8, 9, 10, 11, 12, 13, 14 or more days after the commencement of administration of the anti-HCV therapy); providing a comparison of the first and second levels of gene expression; and determining if the levels of gene expression are sustained (e.g., the levels differ by no more than about 60%, about 50%, about 40%, about 30%, about 20%, about 10%, about 5%, about 2%, or about 1%) between the first and second time points, thereby evaluating the drug candidate.

In some embodiments, the comparison of the first and second levels of gene expression comprises comparing the levels of one or more interferon-sensitive genes (ISG). In some embodiments, the ISG is selected from the group consisting of: IFIT1, RSAD2, IFIT2, IFT16, IFT44, IFIT2, IFIT5, PLSCR1, IFIT3, IFT35, IFITM1, IFITM3, IFT30, IFITM1, IFITM2, GIP2, OAS3, IFIT3, MX1, IFIL44L, IFI27, IFIT2A, PRSAD, or IFITA. In some preferred embodiments, first and second levels of at least 1, 2, 3, 4, 5, 6, 7, 8, 9, or all of: GIP2, OAS3, IFIT3, MX1, IFIL44L, PLSCR1, IFI27, IFIT2A, PRSAD, or IFITA are compared.

In another aspect, the disclosure features a method of evaluating a drug candidate for treatment HCV infection (e.g., chronic HCV) in a subject. The method includes providing a determination of a first level of gene expression associated with HCV infection in the subject at a first time point (e.g., wherein the first time point is prior to, or within about 1, 2, 3, 4, or 5 days of the commencement of, administration of an anti-HCV therapy (e.g., an HCV protease inhibitor, e.g., VX-950)); providing a determination of a second level of gene expression in the subject at a second time point after the first time point and preferably the second time point is after commencement of administration of anti-HCV therapy (e.g., wherein the second time point is taken at least 1, 2, 3, 4, 5, or more days after the first time point or wherein the second time point is 7, 8, 9, 10, 11, 12, 13, 14 or more days after the commencement of administration of the anti-HCV therapy); providing a comparison of the first and second levels of gene expression to a control level of gene expression; and providing a determination of whether there is a smaller difference between the second level and the control level as compared to the difference between the first level and the control level, thereby evaluating a drug candidate.

In some embodiments, the disclosure features a the gene expression associated with HCV infection is determined for a plurality of the genes listed in Table 2. In some embodiments, the plurality includes at least about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, about 95%, about 96%, about 97%, about 98, or about 99% of the genes listed in Table 2. In some embodiments, the plurality includes the genes listed in Table 2.

In some embodiments, the plurality includes a gene from one or more, e.g., each of the following categories (e.g., ontology categories): organismal physiological processes; immune response (e.g., IFIT2, IFIT3, IFIT4, IFI5, IFT16, IFT27, IFT30, IFT35, IFT44, IFITM1, IFITM2, IFITM3, MX1); defense response (e.g., ITGB1); response to biotic stimulus (e.g., CCR1); response to stimulus (e.g., OGG1); response to stress (e.g., CEBP/B); response to pest, pathogen, or parasite (e.g., IFI27); or response to virus (e.g., IRF7, PLSCR1). In some embodiments, the plurality includes a gene from each of 2, 3, 4, 5, 6, 7, or 8 gene ontology categories described herein.

In another aspect, the disclosure features a method of selecting a duration of a protease inhibitor treatment (e.g., treatment with VX-950) for an subject having an HCV infection. The method includes providing an evaluation of whether the patient is an enhanced responder or a non-enhanced responder; and performing at least one of (1) if the subject is an enhanced responder selecting a treatment of a first duration, and (2) if the subject is a non-enhanced responder selecting a second duration of treatment, wherein the first treatment is shorter than the second treatment.

In some embodiments, the patient is a non-enhanced responder and a treatment duration of more than 52, 48, 36, 24, 18, 12, 10, 8, 4 or 2 weeks is selected. In other embodiments, the patient is an enhanced responder and a treatment duration of less than 52, 48, 36, 24, 18, 12, 10, 8, 4 or 2 weeks is selected.

In another aspect, the disclosure features a method of selecting duration of protease inhibitor treatment (e.g., VX-950 treatment) for HCV infection (e.g., chronic HCV) in a subject. The method includes providing a determination of a first level of gene expression associated with HCV infection in the subject at a first time point (e.g., wherein the first time point is prior to, or within about 1, 2, 3, 4, or 5 days of the commencement of, administration of an anti-HCV therapy (e.g., an HCV protease inhibitor, e.g., VX-950)); providing a determination of a second level of gene expression in the subject at a second time point after the first time point and preferably the second time point is after commencement of administration of anti-HCV therapy (e.g., wherein the second time point is taken at least 1, 2, 3, 4, 5, or more days after the first time point or wherein the second time point is 7, 8, 9, 10, 11, 12, 13, 14 or more days after the commencement of administration of the anti-HCV therapy); and providing a comparison of the first and second levels of gene expression and if a sustained level of gene expression (e.g., the levels differ by no more than about 60%, about 50%, about 40%, about 30%, about 20%, about 10%, about 5%, about 2%, or about 1%) is present, selecting a treatment of a first duration, and if a sustained level is not present selecting a second duration of treatment, wherein the first treatment is shorter than the second treatment.

In some embodiments, the first duration is for less than 52, 48, 36, 24, 18, 12, 10, 8, 4 or 2 weeks.

In some embodiments, the second duration is for more than 52, 48, 36, 24, 18, 12, 10, 8, 4 or 2 weeks.

In one aspect, the disclosure features a method evaluating a subject, to determine, e.g., if a subject is an enhanced responder or a non-enhanced responder, to an antiviral treatment, e.g., anti-HCV treatment. The method includes optionally, administering an inhibitor of a viral protease, e.g., VX-950, to the subject; providing a post-administration value for the level of gene expression, (determined, e.g., at the RNA or protein level), for an interferon sensitive gene (ISG) in the subject, providing a comparison of the post administration value with a reference value, (by way of example, the reference value can be the level of expression of the ISG prior to administration of the antiviral treatment), thereby evaluating the subject, e.g., determining if the subject is an enhanced responder or a non-enhanced responder.

In some embodiments, the method includes assigning the subject to a class, and optionally, recording the assignment, e.g., in a computer readable record.

In some embodiments, the evaluation includes determining if the subject is an enhanced responder. In other embodiments, the evaluation includes determining if the subject is a non-enhanced responder.

In some embodiments, the evaluation includes providing information on which to make a decision about the subject (e.g., a decision as to the duration of treatment with an anti-viral agent (e.g., VX-950), or a decision as to which treatment should be administered to a subject, and so forth).

In some embodiments, the method further includes the step of selecting the subject for a preselected treatment.

In some embodiments, the method further includes the step of selecting a duration of treatment of HCV infection (e.g., chronic HCV) in a subject.

In some embodiments, a determination that a subject is an enhanced responder indicates that a shorter duration of treatment can/should/will be/is administered to the subject (e.g., shorter than the treatment which is recommended for a non-enhanced responder, or a duration shorter than currently used with existing anti-viral therapies, e.g., interferon and ribavarin combination therapy, e.g., 52, 48, 36, or 24 weeks), and optionally, that indication is entered into a record.

In some embodiments, a determination that a subject is a non-enhanced responder indicates that a shorter duration of treatment is counter-indicated for the subject (e.g., a duration shorter than currently used with existing anti-viral therapies, e.g., interferon and ribavarin combination therapy, e.g., 52, 48, 36, or 24 weeks), and optionally, that indication is entered into a record.

In some embodiments, providing a comparison of the post administration value with a reference value includes: providing a determination of a post administration level of the ISG in the subject at a first time point (e.g., wherein the first time point is 6, 7, 8, 9, 10, 11, 12, 13, 14 or more days after the commencement of administration of the anti-HCV therapy); providing a determination of a reference value of gene expression associated with HCV infection in the subject at a second time point that is prior to the first time point (e.g., wherein the second time point is prior to, or within about 1, 2, 3, 4, or 5 days of the commencement of, administration of an anti-HCV therapy (e.g., an HCV protease inhibitor, e.g., VX-950)); and providing a comparison of the post administration level and reference value of gene expression, wherein sustained levels of gene expression (e.g., the levels differ by no more than about 60%, about 50%, about 40%, about 30%, about 20%, about 10%, about 5%, about 2%, or about 1%) between the post administration level and reference value indicates that the subject is an enhanced responder.

In some embodiments, the ISG is selected from the group consisting of: IFIT1, RSAD2, IFIT2, IFT16, IFT44, IFIT2, IFIT5, PLSCR1, IFIT3, IFT35, IFITM1, IFITM3, IFT30, IFITM1, IFITM2, GIP2, OAS3, IFIT3, MX1, IFIL44L, IFT27, IFIT2A, PRSAD, or IFITA. In some preferred embodiments, first and second levels of at least 1, 2, 3, 4, 5, 6, 7, 8, 9, or all of: GIP2, OAS3, IFIT3, MX1, IFIL44L, PLSCR1, IFT27, IFIT2A, PRSAD, or IFITA are compared.

In another aspect, the disclosure features a method of predicting treatment outcome for a subject with HCV infection (e.g., chronic HCV). The method includes using a method described herein to determine if a subject is an enhanced responder (e.g., by administering a protease inhibitor, determining a post administration value of gene expression (e.g., for an ISG), and comparing a post-administration value with a reference value) wherein a determination that the subject is an enhanced responder predicts a favorable treatment outcome. In some embodiments, the subject is a human, e.g., a human diagnosed with a viral disorder (e.g., HCV). The disorder can be chronic or acute.

In some embodiments, a viral protease inhibitor is administered to the subject, e.g., the inhibitor of a viral protease (e.g., VX-950) inhibits an HCV protease, e.g., NS3/4A protease. In some embodiments, the inhibitor is VX-950, SCH-503034, or BILN-261 (ciluprevir).

In some embodiments, the disorder is hepatitis C virus infection (e.g., genotype 1, 2, or 3 HCV infection).

In some embodiments, the subject is a human, e.g., a human diagnosed with HCV genotype 1, 2, or 3, a human that has responded well (e.g., succeeded on) or poorly (e.g., failed on) to previous treatments, a human who has previously undergone a particular treatment, a human who has not yet undergone treatment for HCV infection, a human who has been diagnosed as being co-infected with another virus (e.g., hepatitis B and/or HIV).

In some embodiments, the method includes providing a comparison of the post-administration value with a reference value and includes determining if the post-administration value has a predetermined relationship with the reference value, e.g., determining if the post-administration value differs from the reference value by no more than 1, 5, 10, 20, 30, 40, or 50%.

In some embodiments, an ISG is evaluated. In some embodiments, the ISG is selected from the group consisting of: IFIT1, RSAD2, IFIT2, IFI16, IFI44, IFIT2, IFIT5, PLSCR1, IFIT3, IFI35, IFITM1, IFITM3, IFI30, IFITM1, IFITM2, GIP2, OAS3, IFIT3, MX1, IFIL44L, IFI27, IFIT2A, PRSAD, and IFITA. In some embodiments, the ISG is selected from the group consisting of: GIP2, OAS3, IFIT3, MX1, IFIL44L, PLSCR1, IFI27, IFIT2A, PRSAD, and IFITA.

In some embodiments, the reference value is the level of gene expression for the interferon sensitive gene (ISG) in the subject at a first time point (e.g., wherein the first time point is prior to, or within 1, 2, 3, 4, or 5 days of the commencement of, administration of an anti-HCV therapy (e.g., an HCV protease inhibitor, e.g., VX-950)). In some embodiments, the post administration value of the ISG is the level present in the subject at least 1, 2, 3, 4, 5, or more days after the first time point or 7, 8, 9, 10, 11, 12, 13, 14 or more days after the commencement of administration of the anti-HCV therapy. In some embodiments, a subsequent post administration value is determined and the subsequent determination value is the level of the ISG present in the subject 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 days after the post administration value. In some embodiments, the post administration value is a function of the expression of a single ISG In some embodiments, the post administration value is a function of the expression of at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, or 24 ISGs, e.g., selected from the group consisting of: IFIT1, RSAD2, IFIT2, IFT16, IFT44, IFIT2, IFIT5, PLSCR1, IFIT3, IFT35, IFITM1, IFITM3, IFT30, IFITM1, IFITM2, GIP2, OAS3, IFIT3, MX1, IFIL44L, IFT27, IFIT2A, PRSAD, and IFITA. In some embodiments, the post administration value is a function of the expression of at least 2, 3, 4, 5, 6, 7, 8, 9, or 10 ISGs, e.g., selected from the group consisting of: GIP2, OAS3, IFIT3, MX1, IFIL44L, PLSCR1, IFT27, IFIT2A, PRSAD, and IFITA. In some embodiments, the post administration value is a function of the expression of at least 2, but no more than 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, or 24 ISGs, e.g., selected from the group consisting of: IFIT1, RSAD2, IFIT2, IFI16, IFI44, IFIT2, IFIT5, PLSCR1, IFIT3, IFI35, IFITM1, IFITM3, IFI30, IFITM1, IFITM2, GIP2, OAS3, IFIT3, MX1, IFIL44L, IFI27, IFIT2A, PRSAD, and IFITA. In some embodiments, one, two or all of: the post administration value; the reference value, if it is determined from the patient; and the subsequent post administration value, if one is determined, are determined from peripheral blood. In some embodiments, the reference value is a function of: a level determined from the patient and/or a level which is a function of the level determined from one or more other subjects (e.g., a cohort).

In another aspect, the disclosure features a method of selecting a payment class for a course of treatment with a protease inhibitor (e.g., VX-950) for a subject having an HCV infection. The method includes providing (e.g., receiving) an evaluation of whether the patient is an enhanced responder or a non-enhanced responder; and performing at least one of (1) if the subject is an enhanced responder selecting a first payment class, and (2) if the subject is a non-enhanced responder selecting a second payment class.

In some embodiments, assignment of the patient is to the first class and the assignment authorizes payment for a course of treatment for a first duration. In some embodiments, the patient is an enhanced responder and a treatment duration of less than 52, 48, 36, 24, 18, 12, 10, 8, 4 or 2 weeks is authorized.

In some embodiments, assignment of the patient is to the second class and the assignment authorizes payment for a course of treatment for a second duration. In some embodiments, the patient is a non-enhanced responder and a treatment duration of more than 52, 48, 36, 24, 18, 12, 10, 8, 4 or 2 weeks is authorized.

In another aspect, the disclosure features a method of selecting a payment class for a course of treatment with a protease inhibitor (e.g., VX-950) for a subject having an HCV infection. The method includes providing a determination of a first level of gene expression associated with HCV infection in the subject at a first time point (e.g., wherein the first time point is prior to, or within about 1, 2, 3, 4, or 5 days of the commencement of, administration of an anti-HCV therapy (e.g., an HCV protease inhibitor, e.g., VX-950)); providing a determination of a second level of gene expression in the subject at a second time point after the first time point and preferably the second time point is after commencement of administration of anti-HCV therapy (e.g., wherein the second time point is taken at least 1, 2, 3, 4, 5, or more days after the first time point or wherein the second time point is 7, 8, 9, 10, 11, 12, 13, 14 or more days after the commencement of administration of the anti-HCV therapy); and providing a comparison of the first and second levels of gene expression, and if a sustained level of gene expression (e.g., the levels differ by no more than about 60%, about 50%, about 40%, about 30%, about 20%, about 10%, about 5%, about 2%, or about 1%) is present selecting a first payment class, and if a sustained level is not present selecting a second payment class.

In some embodiments, the expression level of one or more interferon-sensitive genes (ISG) is provided. In some embodiments, the ISG is selected from the group consisting of: IFIT1, RSAD2, IFIT2, IFI16, IFI44, IFIT2, IFIT5, PLSCR1, IFIT3, IFI35, IFITM1, IFITM3, IFI30, IFITM1, IFITM2, GIP2, OAS3, IFIT3, MX1, IFIL44L, IFI27, IFIT2A, PRSAD, or IFITA. In some embodiments, the expression level of at least 1, 2, 3, 4, 5, 6, 7, 8, 9, or all of: GIP2, OAS3, IFIT3, MX1, IFIL44L, PLSCR1, IFI27, IFIT2A, PRSAD, or IFITA is provided.

In one aspect, the disclosure features a method of providing information on which to make a decision about a subject, or making such a decision. The method includes providing (e.g., by receiving) an evaluation of a subject, wherein the evaluation was made by a method described herein, e.g., by optionally, administering an inhibitor of a viral protease, e.g., VX-950, to the subject; providing a determination of a post administration level of gene expression for an interferon sensitive gene (ISG) in the subject, thereby providing a post administration value; providing a comparison of the post administration level with a reference value, thereby, providing information on which to make a decision about a subject, or making such a decision.

In some embodiments, the method includes making the decision.

In some embodiments, the method also includes communicating the information to another party (e.g., by computer, compact disc, telephone, facsimile, email, or letter).

In some embodiments, the decision includes selecting a subject for payment, making or authorizing payment for a first course of action if the subject is an enhanced responder and a second course of action if the subject in a non-enhanced responder.

In some embodiments, the decision includes selecting a first course of action if the post administration value has a first predetermined relationship with a reference value, and selecting a second course of action if the post administration value has a second predetermined relationship with the reference value.

In some embodiments, the decision includes selecting a first course of action if the subject is an enhanced responder and a second course of action if the subject in a non-enhanced responder.

In some embodiments, the subject is an enhanced responder and the course of action is authorization of a course of therapy. In some embodiments, the course of therapy is shorter than what is provided to an otherwise similar subject who is a non-enhanced responder, e.g., the course of therapy is less than 52, 48, 36, 24, 18, 12, 10, 8, 4 or 2 weeks.

In some embodiments, the subject is an enhanced responder and the course of action is assigning the subject to a first class. In some embodiments, assignment to the first class will enable payment for a treatment provided to the subject. In some embodiments, payment is by a first party to a second party. In some embodiments, the first party is other than the patient (e.g., subject). In some embodiments, the first party is selected from a third party payor, an insurance company, employer, employer sponsored health plan, HMO, or governmental entity. In some embodiments, the second party is selected from the subject, a healthcare provider, a treating physician, an HMO, a hospital, a governmental entity, or an entity which sells or supplies the drug. In some embodiments, the first party is an insurance company and the second party is selected from the subject, a healthcare provider, a treating physician, an HMO, a hospital, a governmental entity, or an entity which sells or supplies the drug. In some embodiments, the first party is a governmental entity and the second party is selected from the subject, a healthcare provider, a treating physician, an HMO, a hospital, an insurance company, or an entity which sells or supplies the drug.

In some embodiments, the subject is a non-enhanced responder and the course of action is authorization of a course of therapy. In some embodiments, the course of therapy is longer than what is provided to an otherwise similar subject who is an enhanced responder, e.g., the course of therapy is longer than 52, 48, 36, 24, 18, 12, 10, 8, 4 or 2 weeks. In some embodiments, the subject is a non-enhanced responder and the course of action is assigning the subject to a second class. In some embodiments, assignment to the second class will enable payment for a treatment provided to the patient (e.g., subject), e.g., treatment for a period which is longer than a preselected period (e.g., longer than the period of treatment for an enhanced responder). In some embodiments, payment is by a first party to a second party. In some embodiments, the first party is other than the subject. In some embodiments, the first party is selected from a third party payor, an insurance company, employer, employer sponsored health plan, HMO, or governmental entity. In some embodiments, the second party is selected from the subject, a healthcare provider, a treating physician, an HMO, a hospital, a governmental entity, or an entity which sells or supplies the drug. In some embodiments, the first party is an insurance company and the second party is selected from the subject, a healthcare provider, a treating physician, an HMO, a hospital, a governmental entity, or an entity which sells or supplies the drug. In some embodiments, the first party is a governmental entity and the second party is selected from the subject, a healthcare provider, a treating physician, an HMO, a hospital, an insurance company, or an entity which sells or supplies the drug.

In some embodiments, the subject is a human, e.g., a human diagnosed with a viral disorder.

In some embodiments, the inhibitor of a viral protease inhibits an HCV protease, e.g., NS3/4A protease.

In some embodiments, the disorder is chronic or acute.

In some embodiments, the disorder is hepatitis C virus infection (e.g., genotype 1, 2, or 3 HCV infection). In some embodiments, the subject is a human, e.g., a human diagnosed with HCV genotype 1, 2, or 3, a human that has responded well (e.g., succeeded on) or poorly (e.g., failed on) to previous treatments, a human who has previously undergone a particular treatment, a human who has not yet undergone treatment for HCV infection, a human who has been diagnosed as being co-infected with another virus (e.g., hepatitis B and/or HIV).

In some embodiments, comparing the post-administration level with a reference value includes determining if the post-administration level has a predetermined relationship with the reference value, e.g., determining if the post-administration value differs from the reference value by no more than 1, 5, 10, 20, 30, 40, or 50%.

In some embodiments, the inhibitor is VX-950, SCH-503034, or BILN-261 (ciluprevir).

In some embodiments, the ISG is selected from the group consisting of: IFIT1, RSAD2, IFIT2, IFI16, IFI44, IFIT2, IFIT5, PLSCR1, IFIT3, IFI35, IFITM1, IFITM3, IFI30, IFITM1, IFITM2, GIP2, OAS3, IFIT3, MX1, IFIL44L, IFI27, IFIT2A, PRSAD, and IFITA. In some preferred embodiments, the ISG is selected from the group consisting of: GIP2, OAS3, IFIT3, MX1, IFIL44L, PLSCR1, IFT27, IFIT2A, PRSAD, and IFITA.

In some embodiments, the post administration value of the ISG is the level present in the subject at least 1, 2, 3, 4, 5, or more days after the first time point or 7, 8, 9, 10, 11, 12, 13, 14 or more days after the commencement of administration of the anti-HCV therapy.

In some embodiments, a subsequent post administration level is determined and the subsequent determination value is the level of the ISG present in the subject 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 days after the post administration value.

In some embodiments, the post administration value is a function of the expression of a single ISG In some embodiments, the post administration value is a function of the expression of at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, or 24 ISGs, e.g., selected from the group consisting of: IFIT1, RSAD2, IFIT2, IFI16, IFI44, IFIT2, IFIT5, PLSCR1, IFIT3, IFI35, IFITM1, IFITM3, IFI30, IFITM1, IFITM2, GIP2, OAS3, IFIT3, MX1, IFIL44L, IFI27, IFIT2A, PRSAD, and IFITA. In some embodiments, the post administration value is a function of the expression of at least 2, 3, 4, 5, 6, 7, 8, 9, or 10 ISGs, e.g., selected from the group consisting of: GIP2, OAS3, IFIT3, MX1, IFIL44L, PLSCR1, IFI27, IFIT2A, PRSAD, and IFITA. In some embodiments, the post administration value is a function of the expression of at least 2, but no more than 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, or 25 ISGs, e.g., selected from the group consisting of: IFIT1, RSAD2, IFIT2, IFI16, IFI44, IFIT2, IFIT5, PLSCR1, IFIT3, IFI35, IFITM1, IFITM3, IFI30, IFITM1, IFITM2, GIP2, OAS3, IFIT3, MX1, IFIL44L, IFI27, IFIT2A, PRSAD, and IFITA. In some embodiments, the post administration value is a function of the expression of at least 2 ISGs wherein the value is the intrinsic expression value associated with each ISG.

In some embodiments, one, two or all of: the post administration value; the reference value, if it is determined from the patient; and the subsequent post administration value, if one is determined, are determined from peripheral blood.

In some embodiments, the reference value is a function of: a level determined from the patient; and/or a level which is a function of the level determined from one or more other subjects (e.g., a cohort).

In another aspect, the disclosure features a method of selecting a payment class for a course of treatment with a protease inhibitor for a subject having an HCV infection. The method includes identifying the subject as an enhanced responder, and approving, making, authorizing, receiving, transmitting or otherwise allowing payment of a selected course of treatment e.g., a shorter course of treatment (e.g., less than 52, 48, 36, 24, 18, 12, 10, 8, 4 or 2 weeks) than if the subject has been identified as a non-enhanced responder.

In another aspect, the disclosure features a method of selecting a payment class for a course of treatment with a protease inhibitor for a subject having an HCV infection. The method includes identifying the subject as a non-enhanced responder, and approving, making, authorizing, receiving, transmitting or otherwise allowing payment of a selected course of treatment e.g., a longer course of treatment (e.g., more than 52, 48, 36, 24, 18, 12, 10, 8, 4 or 2 weeks) than if the subject had been identified as an enhanced responder.

In one aspect, the disclosure features a method of making a data record. The method includes entering the result of a method described herein into a record, e.g., a computer readable record. In some embodiments, the record is available on the world wide web. In some embodiments, the record is evaluated by a third party payor, an insurance company, employer, employer sponsored health plan, HMO, or governmental entity, or a healthcare provider, a treating physician, an HMO, a hospital, a governmental entity, or an entity which sells or supplies the drug, or is otherwise relied on in a method described herein.

In another aspect, the disclosure features a data record (e.g., computer readable record), wherein the record includes results from a method described herein. In some embodiments, the record is available on the world wide web. In some embodiments, the record is evaluated and/or transmitted to a third party payor, an insurance company, employer, employer sponsored health plan, HMO, or governmental entity, or a healthcare provider, a treating physician, an HMO, a hospital, a governmental entity, or an entity which sells or supplies the drug.

In one aspect, the disclosure features a method of providing data. The method includes providing data described herein, e.g., generated by a method described herein, to provide a record, e.g., a record described herein, for determining if a payment will be provided. In some embodiments, the data is provided by computer, compact disc, telephone, facsimile, email, or letter. In some embodiments, the data is provided by a first party to a second party. In some embodiments, the first party is selected from the subject, a healthcare provider, a treating physician, an HMO, a hospital, a governmental entity, or an entity which sells or supplies the drug. In some embodiments, the second party is a third party payor, an insurance company, employer, employer sponsored health plan, HMO, or governmental entity. In some embodiments, the first party is selected from the subject, a healthcare provider, a treating physician, an HMO, a hospital, an insurance company, or an entity which sells or supplies the drug and the second party is a governmental entity. In some embodiments, the first party is selected from the subject, a healthcare provider, a treating physician, an HMO, a hospital, an insurance company, or an entity which sells or supplies the drug and the second party is an insurance company.

In another aspect, the disclosure features a signature set of probes having a probe for each of the genes in a signature set described herein, e.g., each of a plurality of genes each of which is differentially expressed as between virally infected individuals and non-infected individuals, and contains a sufficient number of differentially expressed genes such that if each of the genes in the signature set is differentially expressed as compared to a non infected reference, it is predictive of infection with no more than about 15, about 10, about 5, about 2.5, or about 1% false positives.

In some embodiments, the signature set of probes includes probes for a plurality of genes listed in Table 2. In some embodiments, the signature set of probes includes probes for at least about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, about 95%, about 96%, about 97%, about 98, or about 99% of the genes listed in Table 2. In some embodiments, the signature set of probes includes probes for the genes listed in Table 2.

In some embodiments, the signature set of probes includes a probe for a gene from one or more, e.g., each of the following categories (e.g., ontology categories): organismal physiological processes; immune response (e.g., IFIT2, IFIT3, IFIT4, IFI5, IFT16, IFT27, IFT30, IFT35, IFT44, IFITM1, IFITM2, IFITM3, MX1); defense response (e.g., ITGB1); response to biotic stimulus (e.g., CCR1); response to stimulus (e.g., OGG1); response to stress (e.g., CEBP/B); response to pest, pathogen, or parasite (e.g., IFI27); or response to virus (e.g., IRF7, PLSCR1). In some embodiments, the signature set of probes includes probes for a gene from each of 2, 3, 4, 5, 6, 7, or 8 of the gene ontology categories.

In some embodiments, the signature set of probes includes probes for one or more interferon-sensitive genes (ISG). In some embodiments, the ISG is selected from the group consisting of: IFIT1, RSAD2, IFIT2, IFI16, IFI44, IFIT2, IFIT5, PLSCR1, IFIT3, IFI35, IFITM1, IFITM3, IFI30, IFITM1, IFITM2, GIP2, OAS3, IFIT3, MX1, IFIL44L, IFI27, IFIT2A, PRSAD, or IFITA. In some preferred embodiments, the signature set of probes includes probes for at least 1, 2, 3, 4, 5, 6, 7, 8, 9, or all of: GIP2, OAS3, IFIT3, MX1, IFIL44L, PLSCR1, IFI27, IFIT2A, PRSAD, or IFITA.

In some embodiments, the signature set of probes includes probes for at least 20, 40, 60, 80, 100, 150, or 200 genes.

In some embodiments, the signature set of probes includes probes for no more than 20, 40, 60, 80, 100, 150, or 200 genes.

In another aspect, the disclosure features a record (e.g., computer readable record) which includes a list and value of expression for each gene represented in the signature set. In some embodiments, the record includes more than one value for each gene, wherein a first value (e.g., pre treatment, e.g., wherein the first value is obtained at a first time point that is prior to, or within 1, 2, 3, 4, or 5 days of the commencement of, administration of an anti-HCV therapy) and a second value (e.g., wherein the second value is obtained post treatment administration, e.g., at least 1, 2, 3, 4, 5, or more days after the first time point or at 7, 8, 9, 10, 11, 12, 13, 14 or more days after the commencement of administration of the anti-HCV therapy) are provided for each gene.

In one aspect, the disclosure features a method of transmitting a record described herein. The method includes a first party transmitting the record to a second party, e.g., by computer, compact disc, telephone, facsimile, email, or letter. In some embodiments, the second party is selected from the subject, a healthcare provider, a treating physician, an HMO, a hospital, a governmental entity, or an entity which sells or supplies the drug. In some embodiments, the first party is an insurance company or government entity and the second party is selected from the subject, a healthcare provider, a treating physician, an HMO, a hospital, a governmental entity, or an entity which sells or supplies the drug. In some embodiments, the first party is a governmental entity or insurance company and the second party is selected from the subject, a healthcare provider, a treating physician, an HMO, a hospital, an insurance company, or an entity which sells or supplies the drug.

In another aspect, the disclosure features an array including a plurality of spatially distinguishable regions, each region having a probe specific for a gene from a signature set of genes described herein, and the array having at least one of the following properties:

if probe specific spatially distinguishable regions for genes other than those in the signature set are present, spatially distinguishable regions for signature set specific probes account for at least 10, 20, 30, 50, 75, 80, 90, 99% of the total probe specific spatially distinguishable regions of the array;

no more than 10, 100, 500, 1,000, 5,000, or 10,000 probe specific spatially distinguishable regions for genes other than those in the signature set are present on the array;

the array is in contact with nucleic acids derived from a subject who has been administered a protease inhibitor, e.g., VX-950, SCH-503034, or BILN-261 (ciluprevir); or

the array is in contact with nucleic acids derived from a subject who has HCV.

In some embodiments, the array includes a duplicate, or triplicate of 1, 5, 10, 20 or all of the regions having a probe specific for a gene from a signature set of genes.

In another aspect, the disclosure features a method of providing data. The method includes providing hybridization data from contacting an array including a plurality of spatially distinguishable regions described herein with a nucleic acid sample derived from a subject (e.g., a subject described herein), and providing a record of such data.

In some embodiments, the subject has an HCV infection.

In some embodiments, the record includes data from hybridizing nucleic acid from the subject prior to administration of a protease inhibitor, e.g., VX-950, to the subject.

In some embodiments, the record includes data from hybridizing nucleic acid from the subject after administration of a protease inhibitor, e.g., VX-950 to the subject.

In some embodiments, the record includes a value which is a function of comparing pre and post administration data.

In another aspect, an evaluation of the ratio of gene expression of ISGs prior to dosing (e.g., with VX-950) in enhanced responders as compared to non-enhanced responders demonstrates that for many ISGs, the pre-dose expression levels are elevated as compared to the levels in non-enhanced responders (see, e.g., Table 5). Thus, the levels of an ISG, e.g., an ISG shown in Table 5 (e.g., IFIT4, IFI44L, RSAD2, IFIT2, IFIT3, IFI16, IFI44, IFIT5, PLSCR1), can be determined for a subject to generate a value that is a function of the ISG level in the subject. This value for the subject can then be compared to a reference value. For example, if the subject's value is compared to a value from an enhanced responder (or cohort of enhanced responders) and the subject's value is similar to this reference value, this can be used to predict that the subject will also be an enhanced responder. If the subject value is compared to a value from a non-enhanced responder (or a cohort of non-enhanced responders) and the subject's value is similar to this reference, this can be used to predict that the subject may not be an enhanced responder. The results of a classification as an enhanced or non-enhanced responder are described herein.

The term “gene expression” as used herein refers to an indicium of levels of gene expression, such as RNA (e.g., mRNA) levels, cDNA levels, and protein levels. The term “gene transcript” as used herein refers to either the full length transcript for a particular gene or to a portion of that transcript (e.g., oligonucleotide, e.g., probe) that allows identification of that portion as corresponding (e.g., specifically) to a particular full length transcript, particular isoform, splice variant or other variant, or polymorphism thereof. Thus, the term “gene transcript” also includes biomarkers of a particular gene transcript, e.g., a biomarker that can be present on a two dimensional array, e.g., gene chip.

A “signature set of genes” as used herein refers to a plurality of gene transcripts, each of which is differentially expressed as between virally (e.g., HCV) infected subjects and non infected subjects and contains a sufficient number of differentially expressed genes such that if each of the genes in the signature set is differentially expressed as compared to a non infected reference (e.g., non infected individual or cohort of non infected individuals), it is predictive of infection in a test subject for whom the presence or absence of infection is being determined. The signature set can be predictive of the presence of infection (e.g., an HCV infection) with no more than about 15%, about 10%, about 5%, about 2.5%, or about 1% false positives. The signature set can have a preset limit for a false discovery rate (e.g., less than about 10%, about 5%, about 2.5%, or about 1%).

As described herein, gene expression can be measured, e.g., by assaying RNA or cDNA levels, or levels of a polypeptide encoded by a given gene transcript.

As used herein, an “interferon-sensitive gene” (ISG) refers to a gene whose expression is affected by interferon signaling, e.g., interferon signaling can cause increased or decreased expression of the ISG. For example, an ISG can have an interferon-stimulated response element (ISRE) in its 5′ upstream region.

As used herein, the term “value” (e.g., determined value, post administration value, reference value) refers to a value that is a function of the level of expression of a gene transcript. For example, a value for a gene can be based on the expression level (e.g., RNA or protein levels) of the gene. The value need not equal a measured expression level. For example, arriving at a value may involve subtracting out background levels, amplifying the level by some determined factor, determining an averaging level from a cohort of subjects, and/or otherwise adjusting the value.

The term “normalization of the signature set” indicates that the signature of a subject varies by less than about 50%, about 40%, about 30%, about 20%, about 10%, about 5%, about 4%, about 3%, about 2%, or about 1% from the signature of a reference (e.g., non-HCV infected subject or cohort of non-HCV infected subjects).

An “enhanced responder”, as used herein, refers to a subject that responds significantly more quickly as compared to a “non-enhanced responder” to anti-viral treatment (e.g., anti-viral protease treatment, e.g., VX-950), in the sense that viral titers decrease significantly more quickly in the enhanced responder. In one embodiment, an enhanced responder will have no more than about 35%, about 50%, about 60%, or about 75% of the viral titer of an otherwise similar non-enhanced responder, where titer can be measured as international units (I.U.) of viral (e.g., HCV) RNA/ml of blood at 14 days after the beginning of treatment. For example, an enhanced responder can have less than or equal to 35 I.U. of HCV RNA/ml at 14 days after the commencement of treatment, while a “non-enhanced responder”, can have greater than or equal to 100 I.U. of HCV RNA/ml at 14 days after the commencement of treatment (e.g., where titers are measured by the COBAS AmpliPrep/COBAS TAQMAN™ HCV Test (Roche Molecular Diagnostics)). Alternatively, an enhanced responder can also be identified by ISG expression. In some embodiments, e.g., in which first and second levels of an ISG are compared, sustained levels of the gene transcript (e.g., the levels differ by no more than about 60%, about 50%, about 40%, about 30%, about 20%, about 10%, about 5%, about 2%, or about 1%) between the first and second time points, e.g., a first time point that is prior to, or within 1, 2, 3, 4, or 5 days of the commencement of, administration of an anti-HCV therapy and the second time point is after commencement of administration of anti-HCV therapy, e.g., wherein the second time point is taken at least 1, 2, 3, 4, 5, or more days after the first time point or wherein the second time point is 7, 8, 9, 10, 11, 12, 13, 14 or more days after the commencement of administration of the anti-HCV therapy, indicate that the subject is an enhanced responder and, e.g., the duration of treatment for the enhanced responder can be shorter than for a non-enhanced responder.

A signature set described herein can be evaluated for specific groups of subjects, e.g., males, females, HCV genotype 1, 2, or 3, particular age groups, races, subjects that have responded well or poorly to previous treatments (e.g., the same or different treatment), subjects who have previously undergone a particular treatment (e.g., the same or different treatment), subjects who have not yet undergone any treatment for HCV infection, subjects who have been diagnosed as being co-infected with another virus (e.g., hepatitis B and/or HIV) and who may or may not have undergone treatment for the other virus, subjects with alcoholic liver disease, etc.

All cited patents, patent applications, and references are hereby incorporated by reference in their entireties. In the case of conflict, the present application controls.

The details of one or more embodiments of the invention are set forth in the accompanying drawings and the description below. Other features, objects, and advantages of the invention will be apparent from the description and drawings, and from the claims.

DESCRIPTION OF DRAWINGS

FIG. 1 is a line graph demonstrating median HCV RNA levels (y axis) over time (x axis) in HCV infected patients after treatment with VX-950 or a placebo control.

FIG. 2 is a graph depicting the correlation of patients receiving VX-950 over time with healthy subject gene expression levels.

FIGS. 3A, 3B, and 3C demonstrate the correlation between sustained levels of IFN-sensitive genes (ISG) and a reduction in plasma HCV RNA levels. FIG. 3A shows mean ratios of IFN-induced gene expression levels (day 14 vs. pre-dose). There is a statistically significant difference in the sustained expression levels of the ISGs. FIG. 3B shows sustained levels of the ISGs in five enhanced responders (left-most bars) who were HCV RNA undetectable at day 14. FIG. 3C shows quantitative real-time PCR confirmation of Affymetrix genechip results. Gene expression modulation of specific ISGs for each of the three groups in FIG. 3B are shown (top left panel shows the results for the enhanced responders while the top right and bottom panels show the results for the non-enhanced responders).

DETAILED DESCRIPTION

The inventors have identified a signature set associated with chronic HCV infection. One or more of the genes of the signature can be used, for example, to diagnose HCV infection, predict the treatment outcome of a subject with HCV, select a treatment regimen, select dosages of a given treatment, evaluate a drug candidate, and/or select the duration of a treatment regimen. The pattern or levels of expression of a plurality of gene transcripts of the signature can correlate with a given treatment regimen or outcome prediction.

Further, the inventors have identified interferon-sensitive genes (ISGs) whose expression levels can change upon HCV infection. For subjects who achieved undetectable plasma HCV status (e.g., enhanced responders), sustained expression of the ISGs was observed, e.g., in peripheral blood (e.g., mononuclear cells). Thus, baseline and/or sustained expression levels of the ISGs can be used to predict treatment outcomes.

Hepatitis C Virus Infection

Hepatitis C: Hepatitis C is a viral infection of the liver and is a major cause of acute hepatitis and chronic liver disease, including cirrhosis and liver cancer. HCV is one of the viruses (A, B, C, D, and E), which together account for the vast majority of cases of viral hepatitis. HCV is an enveloped RNA virus in the faviviridae family which appears to have a narrow host range. Humans and chimpanzees are the only known species susceptible to infection, with both species developing similar disease. An important feature of the virus is the relative mutability of its genome, which may be related to its high propensity (80%) of inducing chronic infection.

The incubation period of HCV infection before the onset of clinical symptoms ranges from 15 to 150 days. In acute infections, the most common symptoms are fatigue and jaundice; however, the majority of cases (between 60% and 70%), even those that develop chronic infection, are asymptomatic. Other symptoms of HCV infection include: dark urine, abdominal pain, loss of appetite, and nausea.

About 80% of newly infected patients progress to develop chronic infection. Cirrhosis develops in about 10% to 20% of persons with chronic infection, and liver cancer develops in 1% to 5% of persons with chronic infection over a period of 20 to 30 years. Most patients suffering from liver cancer who do not have hepatitis B virus infection have evidence of HCV infection. Hepatitis C also exacerbates the severity of underlying liver disease when it coexists with other hepatic conditions. In particular, liver disease progresses more rapidly among persons with alcoholic liver disease and HCV infection.

B cells, monocytes, and dendritic cells take up HCV particles, and degradation of the particles releases viral proteins and dsRNA that activate gene expression in peripheral blood cells. Clearance of plasma HCV RNA and elimination of virus particles can result in normalization of the signature set. Persistence of differential expression, and lack of normalization, of the 258-gene signature set correlates with the presence of HCV RNA, e.g., 2-3 logs of plasma HCV RNA.

Diagnosis: Diagnostic tests for HCV are used to prevent infection through screening of donor blood and plasma, to establish the clinical diagnosis and to make better decisions regarding medical management of a patient. Diagnostic tests commercially available today are based on enzyme immunosorbant assays (EIA) for the detection of HCV specific antibodies. ELIAs can detect more than 95% of chronically infected patients but can detect only 50% to 70% of acute infections.

A recombinant immunoblot assay (RIBA) that identifies antibodies which react with individual HCV antigens can be used as a supplemental test for confirmation of a positive EIA result.

Testing for HCV RNA by amplification methods (e.g., polymerase chain reaction (PCR) or branched DNA assay) can also be utilized for confirmation of serological results as well as for assessing the effectiveness of antiviral therapy. A positive result indicates the presence of active infection and a potential for spread of the infection and or/the development of chronic liver disease.

Genotypes: There are six known genotypes and more than 50 subtypes of HCV, and genotype information is helpful in defining the epidemiology of hepatitis C. Knowing the genotype or serotype (genotype-specific antibodies) of HCV is helpful in making recommendations and counseling regarding therapy. Patients with genotypes 2 and 3 are almost three times more likely than patients with genotype 1 to respond to therapy with alpha interferon or the combination of alpha interferon and ribavirin. Furthermore, when using combination therapy, the recommended duration of treatment depends on the genotype. For patients with genotypes 2 and 3, a 24-week course of combination treatment can be adequate, whereas for patients with genotype 1, a 48-week course is often recommended. For these reasons, testing for HCV genotype is often clinically helpful.

Interferon-Sensitive Genes (ISG)

Interferons (IFN) are classified into two distinct types, designated as type I (IFN-alpha, IFN-beta, IFN-omega, IFN-tau) and type II (IFN-gamma) according to their cellular origin, inducing agents and antigenic and functional properties. Interferons affect the expression of a number of genes following interaction with specific high-affinity plasma membrane receptors. The products of these genes either singly or coordinately mediate the antiviral, growth inhibitory or immunoregulatory activities attributed to IFN. A feature common to most of not all IFN-sensitive genes is the presence of a DNA element which constitutes an IFN-responsive enhancer, usually present in the 5′ upstream region of the genes. This element, termed interferon-stimulated response element (ISRE) binds a nuclear factor(s) translocated from the cytoplasm to the nucleus following IFN-receptor-triggered signal transduction. The binding of these factors to the ISRE represents the initiating event in stimulating RNA-polymerase-II-mediated transcription from IFN-sensitive genes. Depending on the nature of the cells responding to IFN and the genes involved, induced transcription may be prolonged or rapidly terminated. The rapid termination of transcription is dependent in some cases on IFN-induced protein synthesis and also involves factor binding to the ISRE. The ISGs are involved in mediating the antiviral effect of IFN. ISGs include genes that pertain to the functioning of immune cells, including genes involved in antigen processing and presentation, T-cell activation, lymphocyte trafficking, and effector functions. The ISGs can enhance immunity against viruses, e.g., HCV. Examples of ISGs are listed in Table 5.

Sustained expression of ISGs was seen in subjects who cleared plasma HCV RNA. This can reflect restored intrinsic antiviral defenses and secretion of interferons, and may be a sign of re-emergence of an effective immune response that is essential to eliminate residual HCV infected hepatocytes. Expression of ISGs and other genes associated with acquired immunity may be monitored to establish potential correlations with, and to make predictions of, treatment outcomes. Further, gene or protein therapy with an ISG (e.g., an ISG listed in Table 5), can be used alone or as part of an anti-viral (e.g., anti-HCV) therapy, e.g., gene or protein therapy with an ISG can be used in combination with an anti-viral agent, e.g., an HCV protease inhibitor, e.g., VX-950, SCH-503034, or BILN-261 (ciluprevir).

Treatment of HCV

Antiviral drugs such as interferon taken alone or in combination with ribavirin, can be used for the treatment of persons with chronic hepatitis C. Treatment with interferon (or pegylated interferon) (e.g., interferon-alpha) alone is effective in about 10% to 20% of patients. Interferon (or pegylated interferon) combined with ribavirin is effective in about 30% to 50% of patients. Additional treatments include VX-950, either alone or in combination with interferon (or pegylated interferon) and/or ribavarin, or another anti-viral or immunomodulatory agent.

There is no vaccine against HCV. Research is in progress but the high mutability of the HCV genome complicates vaccine development.

The inventions described herein can be used as part of the evaluation of a subject with HCV and/or in the selection of a suitable treatment regimen, e.g., VX-950 alone or in combination with another agent, or another therapy (e.g., another monotherapy or combination therapy) described herein. For example, the methods and reagents described herein can be used to select a treatment regimen for a subject, e.g., a subject that has been identified as being an enhanced responder or non-enhanced responder.

VX-950

VX-950 is a competitive, reversible peptidomimetic HCV NS3/4A protease inhibitor with a steady state binding constant (ki*) of 3 nM (and with a Ki of 8 nM) and is described in International Application WO 02/018369.

The structure of VX-950 is:

VX-950 is highly insoluble in water. VX-950 may be prepared by methods known to those skilled in the art (see, e.g., International Applications WO 02/18369 and WO 2005/123076; U.S. application Ser. No. 11/147,524 (filed Jun. 8, 2005)). VX-950 can be formulated into tablets, as described in U.S. App. Nos. 60/764,654 (filed Feb. 2, 2006), 60/784,427 (filed Mar. 20, 2006), 60/784,428 (filed Mar. 20, 2006), 60/784,275 (filed Mar. 20, 2006), Ser. No. 11/687,716 (filed Mar. 10, 2007), Ser. No. 11/687,779 (filed Mar. 19, 2007), PCT App. No. PCT/US2007/061456 (filed Feb. 1, 2007).

Inhibition of NS3/4A by VX-950 can restore IFN signaling and block viral replication in hepatocytes and cleavage of TRIF/CARDIF, thereby restoring IRF3 and RIG-1 signaling and transcription of ISGs that can activate intrinsic anti-viral defenses, including production of IFNβ, in hepatocytes.

Treatment with VX-950

VX-950 Monotherapy: Dosage levels of from about 0.01 to about 100 mg/kg body weight per day, preferably from about 10 to about 100 mg/kg body weight per day of VX-950 are useful for the prevention and treatment of HCV mediated disease. In some embodiments, dosage levels from about 0.4 to about 10 g/day, for example from about 1 to about 4 g/day, preferably from about 2 to about 3.5 g/day, per person (based on the average size of a person calculated at about 70 kg) are included. Typically, the pharmaceutical compositions of, and according to, this invention will be administered from about 1 to about 5 times per day, preferably from about 1 to about 3 times per day, or alternatively, as a continuous infusion. In some embodiments, VX-950 is administered using a controlled release formulation. In some embodiments, this can help to provide relatively stable blood levels of VX-950.

In some embodiments, amorphous VX-950 is administered. The dose of amorphous VX-950 can be a standard dose, e.g., about 1 g to about 5 g a day, more preferably about 2 g to about 4 g a day, more preferably about 2 g to about 3 g a day, e.g., about 2.25 g or about 2.5 g a day. For example, a does of about 450 mg, 750 mg, or 1250 mg can be administered to a subject three times a day. A dose of 1250 mg can be given twice daily. For example, a dose of about 2.25 g/day of amorphous VX-950 can be administered to a patient, e.g., about 750 mg administered three times a day. Such a dose can be administered, e.g., as three 250 mg doses three times a day or as two 375 mg doses three times a day. In some embodiments, the 250 mg dose is in an about 700 mg tablet. In some embodiments, the 375 mg dose is in an about 800 mg tablet. As another example, a dose of about 2.5 g/day of amorphous VX-950 can be administered to a patient, e.g., about 1250 mg administered two times a day. As another example, about 1 g to about 2 g of amorphous VX-950 a day can be administered to a patient, e.g., about 1.35 g of amorphous VX-950 can be administered to a patient, e.g., about 450 mg administered three times a day. The dose of amorphous VX-950 can be administered e.g., as a spray dried dispersion or as a tablet (e.g., a tablet that comprises VX-950, e.g., in a spray dried dispersion).

In some embodiments, the solid (e.g., spray dried) dispersions of VX-950 described herein contain at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85% or greater of VX-950 (e.g., amorphous VX-950). Because these dispersions can include greater amounts of VX-950 for a given amount of a dispersion (e.g., a greater percent by weight of VX-950), for the same amount by weight of solid dispersion, a greater amount of VX-950 can be incorporated into a pharmaceutical composition, thereby increasing the load of the active ingredient in that composition. As a result, a subject receiving VX-950 can take fewer doses of VX-950 and yet intake the same amount of drug. For example, to receive a dose of 750 mg of VX-950, a subject can take two 375 mg doses of VX-950 containing a solid dispersion described herein instead of three 250 mg doses. This can be an improvement or a preferred dose for some patients. As another example, the increased load of amorphous VX-950 in a solid dispersion can allow administration of a larger dose of VX-950 to a subject in a fixed total dose of a pharmaceutical composition (e.g., a tablet of a standard size may contain a larger percentage (and thereby dose) of amorphous VX-950). Conversely, the increased load of amorphous VX-950 can allow a fixed dose amount of amorphous to be administered to a subject in a small total dose of a pharmaceutical composition (e.g., a standard dose of amorphous VX-950 can be administered in a smaller tablet).

In some embodiments, the amorphous VX-950 is not 100% potent or pure (e.g., the potency or purity is at least about 90%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% potent), in which case the doses described above refer to the amount of potent or pure VX-950 administered to a patient rather than the total amount of VX-950. These doses can be administered to a patient as a monotherapy and/or as part of a combination therapy, e.g., as described further below.

Such administration can be used as a chronic or acute therapy. The amount of active ingredient that may be combined with the carrier materials to produce a single dosage form will vary depending upon the subject treated and the particular mode of administration. A typical preparation will contain from about 5% to about 95% active compound (w/w). Preferably, such preparations contain from about 20% to about 80%, from about 25% to about 70%, from about 30% to about 60% active compound.

When the compositions or methods of this disclosure involve a combination of VX-950 and one or more additional therapeutic or prophylactic agents, both the compound and the additional agent should be present at dosage levels of between about 10 to 100%, and more preferably between about 10 to 80% of the dosage normally administered in a monotherapy regimen.

Upon improvement of a patient's condition, a maintenance dose of a compound, composition or combination of this disclosure may be administered, if necessary. Subsequently, the dosage or frequency of administration, or both, may be reduced, e.g., to about ½ or ¼ or less of the dosage or frequency of administration, as a function of the symptoms, to a level at which the improved condition is retained when the symptoms have been alleviated to the desired level, treatment should cease. Patients may, however, require intermittent treatment on a long-term basis upon any recurrence of disease symptoms.

It should also be understood that a specific dosage and treatment regimen for any particular patient will depend upon a variety of factors, including the activity of the specific compound employed, the age, body weight, general health, sex, diet, time of administration, rate of excretion, drug combination, influence of any previous therapies undergone by the subject, and the judgment of the treating physician and the severity of the particular disease being treated. The amount of active ingredients will also depend upon the particular described compound and the presence or absence and the nature of the additional anti-viral agent in the composition.

Combination Therapy

More than one therapeutic agent can be used to treat HCV.

In some embodiments, two or more agents to treat HCV can be started at the same time or within 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14 or more days of each other, or optionally, can be administered sequentially. In combination therapy, the course of the first and second agents can be the same, can overlap but be different, or can be sequential, e.g., the course of the first agent is given and then a course of the second agent is given. In a preferred embodiment, therapeutic levels of both agents are present for at least a portion of the therapy.

In some embodiments, a protease inhibitor, e.g., VX-950, is administered to a subject and ISG (e.g., one or more of the ISGs described herein) expression is measured. In some embodiments, ISG expression is measured prior to, or within about 1, 2, 3, 4, or 5, days of the commencement of, administration of the protease inhibitor (first time point) and/or at least 1, 2, 3, 4, 5, or more days after the first time point or at least 7, 8, 9, 10, 11, 12, 13, 14 or more days after the commencement of the protease inhibitor, and optionally at another time point. If ISG expression is measured at more than one time point, the levels of ISG expression can be compared. For example, if ISG levels are sustained at the two time points, the subject can be classified as an enhanced responder; if ISG levels are not sustained, the subject can be classified as a non-enhanced responder, as described herein. The classification of the subject can be used to decide a treatment regimen, as described herein. After the ISG level is measured at one or more time points, a second therapy (e.g., while continuing with the first treatment with the protease inhibitor) can optionally be started, e.g., interferon, ribavarin, a second protease inhibitor, or other therapy described herein, can be administered to the subject. The second therapy can be initiated within about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14 or more days of the initiation of the first therapy. The second therapy can be maintained for the duration of the treatment of the first therapy, or for a longer or shorter period than the period used for the first therapy. For example, the second therapy can be administered at a dose and for a duration previously known for that therapy (e.g., peg-interferon or ribavarin).

Examples of agents that can be used to treat HCV infection, alone or in combination therapies (e.g., with another agent described therein or with VX-950), are described in International Publication WO 02/18369. The combinations specifically recited therein can be combined with methods described herein. The methods and reagents described herein can be used to select a treatment regimen (e.g., a combination therapy) for a subject, e.g., a subject that has been identified as being an enhanced responder or non-enhanced responder.

VX-950 Combination Therapy: VX-950 can optionally be administered with another component comprising an additional agent, e.g., selected from an immunomodulatory agent; an antiviral agent; an inhibitor of HCV protease; an inhibitor of another target in the HCV life cycle; an inhibitor of internal ribosome entry; a broad-spectrum viral inhibitor; a cytochrome P-450 inhibitor(s); or combinations thereof.

Accordingly, in another embodiment, this invention provides a method comprising administering any form of VX-950, any solid dispersion, or any composition according to this invention, a CYP inhibitor, and another anti-viral agent, preferably an anti-HCV agent. Such anti-viral agents include, but are not limited to, immunomodulatory agents, such as α-, β-, and γ-interferons, pegylated derivatized interferon-α compounds, and thymosin; other anti-viral agents, such as ribavirin, amantadine, and telbivudine; other inhibitors of hepatitis C proteases (NS2-NS3 inhibitors and NS3/NS4A inhibitors); inhibitors of other targets in the HCV life cycle, including helicase, polymerase, and metalloprotease inhibitors; inhibitors of internal ribosome entry; broad-spectrum viral inhibitors, such as IMPDH inhibitors (e.g., compounds of U.S. Pat. Nos. 5,807,876, 6,498,178, 6,344,465, 6,054,472; International Applications WO 97/40028, WO 98/40381, WO 00/56331, and mycophenolic acid and derivatives thereof, and including, but not limited to VX-497, VX-148, and/or VX-944); or combinations of any of the above.

A preferred combination therapy comprises a formulation of amorphous VX-950 described herein and interferon-α, e.g., pegylated derivatized interferon-α (e.g., pegylated interferon-alpha-2a; e.g., PEGASYS®, e.g., at its standard dose). For example, a dose (e.g., as described above) of amorphous VX-950, e.g., about 2 g to about 3 g (e.g., 2.5 g, 2.25 g (e.g., 750 mg three times a day)), e.g., in the form of a tablet described herein can be administered three times a day and pegylated interferon-alpha-2a can be administered at a standard dose, e.g., 180 μg once weekly by subcutaneous administration, e.g., for 48 or 52 weeks. As another example, VX-950 can be administered with both pegylated interferon-alpha-2 and ribavirin. For example, about 2 g to about 3 g (e.g., about 2.5 g, about 2.25 g (e.g., 750 mg three times a day)) of amorphous VX-950 in the form of a tablet described herein, can be administered three times a day in combination with 180 μg of pegylated interferon-alpha-2a (e.g., PEGASYS®) once a week and ribavirin (e.g., COPEGUS®; (1-beta-D-ribofuranosyl-1H-1,2,4-triazole-3-carboxamide, available from ICN Pharmaceuticals, Inc., Costa Mesa, Calif.; described in the Merck Index, entry 8365, Twelfth Edition) at 1000-1200 mg/day, e.g., for 48 or 52 weeks, for genotype 1 patients, or in combination with 180 μg of pegylated interferon-alpha-2a once a week plus ribavirin at 800 mg/day for patients with genotype 2 or 3 hepatitis C.

Other agents that can be used in combination with VX-950 include those described in various published U.S. patent applications. These publications provide additional teachings of compounds and methods that could be used in combination with VX-950 in the methods of this invention, particularly for the treatment of hepatitis. It is contemplated that any such methods and compositions may be used in combination with the methods and compositions of the present invention. For brevity, the disclosure the disclosures from those publications is referred to by reference to the publication number. Exemplary such publications include U.S. Pub. Nos. 20040058982; 20050192212; 20050080005; 20050062522; 20050020503; 20040229818; 20040229817; 20040224900; 20040186125; 20040171626; 20040110747; 20040072788; 20040067901; 20030191067; 20030187018; 20030186895; 20030181363; 20020147160; 20040082574; 20050192212; 20050187192; 20050187165; 20050049220; and US20050222236.

Additional examples of agents include, but are not limited to, ALBUFERON™ (albumin-Interferon alpha) available from Human Genome Sciences; PEG-INTRON® (peginterferon alfa-2b, available from Schering Corporation, Kenilworth, N.J.); INTRON-Ag, (VIRAFERON®, interferon alfa-2b available from Schering Corporation, Kenilworth, N.J.); REBETROL®(Schering Corporation, Kenilworth, N.J.); COPEGUS®(Hoffmann-La Roche, Nutley, N.J.); PEGASYS®(peginterferon alfa-2a available Hoffmann-La Roche, Nutley, N.J.); ROFERON®(recombinant interferon alfa-2a available from Hoffmann-La Roche, Nutley, N.J.); BEREFOR®(interferon alfa 2 available from Boehringer Ingelheim Pharmaceutical, Inc., Ridgefield, Conn.); SUMIFERON®(a purified blend of natural alpha interferons such as Sumiferon available from Sumitomo, Japan); WELLFERON®(interferon alpha n1 available from Glaxo Wellcome Ltd., Great Britain); ALFERON® (a mixture of natural alpha interferons made by Interferon Sciences, and available from Purdue Frederick Co., CT); α-interferon; natural alpha interferon 2a; natural alpha interferon 2b; pegylated alpha interferon 2a or 2b; consensus alpha interferon (Amgen, Inc., Newbury Park, Calif.); REBETRON® (Schering Plough, Interferon-alpha 2B+Ribavirin); pegylated interferon alpha (Reddy, K. R. et al. “Efficacy and Safety of Pegylated (40-kd) Interferon alpha-2a Compared with Interferon alpha-2a in Noncirrhotic Patients with Chronic Hepatitis C (Hepatology, 33, pp. 433-438 (2001); consensus interferon (INFERGEN®) (Kao, J. H., et al., “Efficacy of Consensus Interferon in the Treatment of Chronic Hepatitis” J. Gastroenterol. Hepatol. 15, pp. 1418-1423 (2000); lymphoblastoid or “natural” interferon; interferon tau (Clayette, P. et al., “IFN-tau, A New Interferon Type I with Antiretroviral activity” Pathol. Biol. (Paris) 47, pp. 553-559 (1999); interleukin-2 (Davis, G. L. et al., “Future Options for the Management of Hepatitis C.” Seminars in Liver Disease, 19, pp. 103-112 (1999); Interleukin-6 (Davis et al. “Future Options for the Management of Hepatitis C.” Seminars in Liver Disease 19, pp. 103-112 (1999); interleukin-12 (Davis, G. L. et al., “Future Options for the Management of Hepatitis C.” Seminars in Liver Disease, 19, pp. 103-112 (1999); and compounds that enhance the development of type 1 helper T cell response (Davis et al., “Future Options for the Management of Hepatitis C.” Seminars in Liver Disease, 19, pp. 103-112 (1999)). Also included are compounds that stimulate the synthesis of interferon in cells (Tazulakhova, E. B. et al., “Russian Experience in Screening, analysis, and Clinical Application of Novel Interferon Inducers” J. Interferon Cytokine Res., 21 pp. 65-73) including, but are not limited to, double stranded RNA, alone or in combination with tobramycin, and Imiquimod (3M Pharmaceuticals; Sauder, D. N. “Immunomodulatory and Pharmacologic Properties of Imiquimod” J. Am. Acad. Dermatol., 43 pp. S6-11 (2000). In addition, known protease inhibitors (e.g., HCV protease inhibitors) can be tested for suitability with the methods described herein.

Each agent may be formulated in separate dosage forms. Alternatively, to decrease the number of dosage forms administered to a patient, each agent may be formulated together in any combination. For example, the VX-950 may be formulated in one dosage form and any additional agents may be formulated together or in another dosage form. VX-950 can be dosed, for example, before, after, or during the dosage of the additional agent.

A method according to this invention may also comprise the step of administering a cytochrome P450 monooxygenase (CYP) inhibitor. CYP inhibitors may be useful in increasing liver concentrations and/or increasing blood levels of compounds (e.g., VX-950) that are inhibited by CYP.

The advantages of improving the pharmacokinetics of a drug (e.g., by administering a CYP inhibitor) are well accepted in the art. By administering a CYP inhibitor, this invention provides for decreased metabolism of the protease inhibitor, VX-950. The pharmacokinetics of the protease inhibitor are thereby improved. The advantages of improving the pharmacokinetics of a drug are well accepted in the art. Such improvement may lead to increased blood levels of the protease inhibitor. More importantly for HCV therapies, the improvement may lead to increased concentrations of the protease inhibitor in the liver.

In a method of this invention, the amount of CYP inhibitor administered is sufficient to increase the blood levels of the VX-950 as compared to the blood levels of this protease inhibitor in the absence of a CYP inhibitor. Advantageously, in a method of this invention, an even further lower dose of protease inhibitor may be therefore used (relative to administration of a protease inhibitor alone).

Accordingly, another embodiment of this invention provides a method for increasing blood levels or increasing liver concentrations of VX-950 in a patient receiving VX-950 comprising administering to the patient a therapeutically effective amount of VX-950 and a cytochrome P450 monooxygenase inhibitor.

In addition to treating patients infected with hepatitis C, the methods of this invention may be used to prevent a patient from becoming infected with hepatitis C, e.g., a patient who may undergo a blood transfusion. Accordingly, one embodiment of this invention provides a method for preventing a hepatitis C virus infection in a patient (e.g., prophylactic treatment) comprising administering to the patient a) a formulation of VX-950 or any composition according to this invention; and optionally, b) a cytochrome P450 monooxygenase inhibitor.

As would be realized by skilled practitioners, if a method of this invention is being used to treat a patient prophylactically, and that patient becomes infected with hepatitis C virus, the method may then treat the infection. Therefore, one embodiment of this invention provides VX-950 or any composition according to this invention and optionally, a cytochrome P450 monooxygenase inhibitor, wherein the inhibitors of the combination are in therapeutically effective amounts for treating or preventing a hepatitis C infection in a patient.

If an embodiment of this invention involves a CYP inhibitor, any CYP inhibitor that improves the pharmacokinetics of VX-950 may be used in a method of this invention. These CYP inhibitors include, but are not limited to, ritonavir (International Application WO 94/14436), ketoconazole, troleandomycin, 4-methylpyrazole, cyclosporin, clomethiazole, cimetidine, itraconazole, fluconazole, miconazole, fluvoxamine, fluoxetine, nefazodone, sertraline, indinavir, nelfinavir, amprenavir, fosamprenavir, saquinavir, lopinavir, delavirdine, erythromycin, VX-944, and VX-497. Preferred CYP inhibitors include ritonavir, ketoconazole, troleandomycin, 4-methyl pyrazole, cyclosporin, and clomethiazole. For preferred dosage forms of ritonavir, see U.S. Pat. No. 6,037,157, and the documents cited therein: U.S. Pat. No. 5,484,801, U.S. application Ser. No. 08/402,690, and International Applications WO 95/07696 and WO 95/09614).

The structure of VX-944 is as follows:

VX-497 is an IMPDH inhibitor. A combination of VX-497, pegylated interferon-α (IFN-α), and ribavirin is currently in clinical development for treating HCV (W. Markland et al., (2000) Antimicrobial & Antiviral Chemotherapy, 44, p. 859; U.S. Pat. No. 6,541,496).

The structure of VX-497 is as follows:

Methods for measuring the ability of a compound to inhibit cytochrome P450 monooxygenase activity are known (see U.S. Pat. No. 6,037,157 and Yun, et al. (1993) Drug Metabolism & Disposition, vol. 21, pp. 403-407).

A CYP inhibitor employed in this invention may be an inhibitor of only one isozyme or more than one isozyme. If the CYP inhibitor inhibits more than one isozyme, the inhibitor may nevertheless inhibit one isozyme more selectively than another isozyme. Any such CYP inhibitors may be used in a method of this invention.

In a method of this invention, the CYP inhibitor may be administered together with a formulation of VX-950 or any composition according to this invention in the same dosage form or in separate dosage forms.

If the CYP inhibitor and the other components of the combination are administered in separate dosage forms, each inhibitor may be administered about simultaneously. Alternatively, the CYP inhibitor may be administered in any time period around administration of the combination. That is, the CYP inhibitor may be administered prior to, together with, or following each component of the combination. The time period of administration should be such that the CYP inhibitor affects the metabolism of a component of the combination, preferably, of VX-950. For example, if VX-950 is administered first, the CYP inhibitor should be administered before VX-950 is substantially metabolized and/or excreted (e.g., within the half-life of VX-950).

Nucleic Acid and Protein Analysis

The genes (or their encoded polypeptides) of a signature set described herein can be used in the diagnosis of HCV, and/or in predicting the treatment outcome of a subject with HCV. Further, the levels of one or more (or all) genes (or encoded polypeptide) of the signature can be used to select a treatment regimen, select dosages of a given treatment, and/or select the duration of a treatment regimen. For example, the levels of an ISG at two or more time points (e.g., prior to treatment or within 1, 2, 3, 4, or 5 days of starting treatment and at another time(s), e.g., at least 1, 2, 3, 4, 5, or more days after the first time point or 7, 8, 9, 10, 11, 12, 13, 14 or more days after the start of treatment) can be used to predict a subject's response to a given therapy (e.g., VX-950). As another example, the pattern or levels of expression of a plurality of genes (e.g., an ISG(s)) can correlate with a given treatment regimen or outcome prediction.

Numerous methods for detecting expression of a gene (e.g., a nucleic acid and/or encoded protein of one or more genes of the signature set described herein) (e.g., an ISG), and for detecting the levels of expression, are available to the skilled artisan. The methods include hybridization-based methods for nucleic acid detection (e.g., PCR or Northern blot), and antibody-based methods for protein detection (e.g., Western blot, radioimmunoassay (RIA), or ELISA).

The expression levels of a gene of the signature set can be determined using nucleic acid or hybridization or amplification techniques known in the art (e.g., using PCR or Northern blot). The expression levels in a sample (e.g., from a subject with hepatitis C) can be quantitatively or qualitatively compared to the levels in a reference or control (e.g., the levels in a healthy subject).

Arrays are particularly useful molecular tools for characterizing a sample, e.g., a sample from a subject, e.g., a subject with hepatitis C. For example, an array having capture probes for multiple genes (or for multiple proteins), including probes for a gene(s) of the signature set described herein, can be used in a method described herein. Altered expression of a nucleic acid and/or encoded protein of the signature set described herein can be used to evaluate a sample, e.g., a sample from a subject, e.g., to predict the subject's response to treatment (e.g., treatment with VX-950).

Arrays can have many addresses, e.g., locatable sites, on a substrate. The featured arrays can be configured in a variety of formats, non-limiting examples of which are described below. The substrate can be opaque, translucent, or transparent. The addresses can be distributed, on the substrate in one dimension, e.g., a linear array; in two dimensions, e.g., a planar array; or in three dimensions, e.g., a three dimensional array. The solid substrate may be of any convenient shape or form, e.g., square, rectangular, ovoid, or circular.

Arrays can be fabricated by a variety of methods, e.g., photolithographic methods (see, e.g., U.S. Pat. Nos. 5,143,854; 5,510,270; and 5,527,681), mechanical methods (e.g., directed-flow methods as described in U.S. Pat. No. 5,384,261), pin based methods (e.g., as described in U.S. Pat. No. 5,288,514), and bead based techniques (e.g., as described in PCT US/93/04145).

The capture probe can be a single-stranded nucleic acid, a double-stranded nucleic acid (e.g., which is denatured prior to or during hybridization), or a nucleic acid having a single-stranded region and a double-stranded region. Preferably, the capture probe is single-stranded. The capture probe can be selected by a variety of criteria, and preferably is designed by a computer program with optimization parameters. The capture probe can be selected to hybridize to a sequence rich (e.g., non-homopolymeric) region of the gene. The T_mof the capture probe can be optimized by prudent selection of the complementarity region and length. Ideally, the T_mof all capture probes on the array is similar, e.g., within 20, 10, 5, 3, or 2° C. of one another.

The isolated nucleic acid is preferably mRNA that can be isolated by routine methods, e.g., including DNase treatment to remove genomic DNA and hybridization to an oligo-dT coupled solid substrate (e.g., as described in Current Protocols in Molecular Biology, John Wiley & Sons, N.Y). The substrate is washed, and the mRNA is eluted.

The isolated mRNA can be reversed transcribed and optionally amplified, e.g., by rtPCR (e.g., as described in U.S. Pat. No. 4,683,202). The nucleic acid can be an amplification product, e.g., from PCR (U.S. Pat. Nos. 4,683,196 and 4,683,202); rolling circle amplification (“RCA,” U.S. Pat. No. 5,714,320), isothermal RNA amplification or NASBA (U.S. Pat. Nos. 5,130,238; 5,409,818; and 5,554,517), and strand displacement amplification (U.S. Pat. No. 5,455,166). The nucleic acid can be labeled during amplification, e.g., by the incorporation of a labeled nucleotide. Examples of preferred labels include fluorescent labels, e.g., red-fluorescent dye Cy5 (Amersham) or green-fluorescent dye Cy3 (Amersham), and chemiluminescent labels, e.g., as described in U.S. Pat. No. 4,277,437. Alternatively, the nucleic acid can be labeled with biotin, and detected after hybridization with labeled streptavidin, e.g., streptavidin-phycoerythrin (Molecular Probes).

The labeled nucleic acid can be contacted to the array. In addition, a control nucleic acid or a reference nucleic acid can be contacted to the same array. The control nucleic acid or reference nucleic acid can be labeled with a label other than the sample nucleic acid, e.g., one with a different emission maximum. Labeled nucleic acids can be contacted to an array under hybridization conditions. The array can be washed, and then imaged to detect fluorescence at each address of the array. The levels of expression in the control and sample nucleic acids can be compared relative to each other or to a reference value.

The expression level of a polypeptide encoded by a gene of the signature set can be determined using an antibody specific for the polypeptide (e.g., using a Western blot or an ELISA). The polypeptide levels in a sample (e.g., from a subject with hepatitis C) can be quantitatively or qualitatively compared to the levels in a reference or control (e.g., the levels in a healthy subject).

Moreover, the expression levels of multiple proteins, such as a plurality of the gene transcripts of the signature set provided herein, can be rapidly determined in parallel using a polypeptide array having antibody capture probes for each of the polypeptides. Antibodies specific for a polypeptide can be generated as generally known in the art. The polypeptide level of a gene transcript provided herein (e.g., an ISG) can be measured in a biological sample from a subject (e.g., blood, serum, or plasma).

A low-density (96 well format) protein array has been developed in which proteins are spotted onto a nitrocellulose membrane (Ge (2000) Nucleic Acids Res. 28, e3, I-VII). A high-density protein array (100,000 samples within 222×222 mm) used for antibody screening was formed by spotting proteins onto polyvinylidene difluoride (PVDF) (Lueking et al. (1999) Anal. Biochem. 270:103-111). See also, e.g., Mendoza et al. (1999). Biotechniques 27:778-788; MacBeath and Schreiber (2000) Science 289:1760-1763; and De Wildt et al. (2000) Nature Biotech. 18:989-994. These art-known methods and others can be used to generate an array of antibodies for detecting the abundance of polypeptides (e.g., encoded by gene transcripts of the signature set) in a sample. The sample can be labeled, e.g., biotinylated, for subsequent detection with streptavidin coupled to a fluorescent label. The array can then be scanned to measure binding at each address. The amount of binding in a sample can be compared to the amount of binding in a control or reference.

The nucleic acid and polypeptide arrays of the invention can be used in wide variety of applications. For example, the arrays can be used to analyze a sample from a subject (e.g., peripheral blood or tissue from a liver biopsy). The sample is compared to data obtained previously, e.g., known clinical specimens, other patient samples, a healthy (non-infected) control, or data obtained from a cohort of subjects. Further, the arrays can be used to characterize a cell culture sample, e.g., to determine a cellular state after varying a parameter, e.g., dosing a patient with an anti-HCV therapy, e.g., VX-950.

The expression data can be stored in a database, e.g., a relational database such as a SQL database (e.g., Oracle or Sybase database environments). The database can have multiple tables. For example, raw expression data can be stored in one table, wherein each column corresponds to a gene (e.g., a gene transcript of the signature) being assayed, e.g., an address or an array, and each row corresponds to a sample. A separate table can store identifiers and sample information, e.g., the batch number of the array used, date, and other quality control information.

Expression profiles obtained from gene expression analysis on an array can be used to compare samples and/or cells in a variety of states as described in Golub et al. ((1999) Science 286:531). In one embodiment, expression (e.g., mRNA expression or protein expression) information for a gene transcript provided herein are evaluated, e.g., by comparison a reference value, e.g., a control value from a healthy subject. Reference values can also be obtained from statistical analysis, e.g., to provide a reference value for a cohort of subjects, e.g., age and gender matched subjects, e.g., normal subjects or subjects who have HCV, e.g., a particular HCV genotype or who have undergone a particular HCV therapy. Statistical similarity to a particular reference (e.g., to a reference for a risk-associated cohort) or a normal cohort can be used to provide an assessment (e.g., a prediction of treatment outcome) to a subject, e.g., a subject who has been diagnosed with HCV.

Subjects suitable for treatment can also be evaluated for expression and/or activity of a gene transcript of the signature set. Subjects can be identified as suitable for treatment (e.g., with VX-950 dosing), if the expression and/or activity for a particular gene transcript is elevated relative to a reference, e.g., reference value, e.g., a reference value associated with normal.

Subjects who are being administered an agent described herein (e.g., VX-950) or other treatment can be evaluated as described for expression and/or activity of a gene(s) described herein. The subject can be evaluated at multiple times, e.g., at multiple times during a course of therapy, e.g., during a therapeutic regimen, and/or prior to commencement of the regimen. Treatment of the subject can be modified depending on how the subject is responding to the therapy. For example, a change in a gene's expression or activity (e.g., normalization of the signature) can be indicative of responsiveness.

Particular effects mediated by an agent may show a difference (e.g., relative to an untreated subject, control subject, or other reference) that is statistically significant (e.g., P value<0.05 or 0.02). Statistical significance can be determined by any art known method. Exemplary statistical tests include: the Students T-test, Mann Whitney U non-parametric test, and Wilcoxon non-parametric statistical test. Some statistically significant relationships have a P value of less than 0.05 or 0.02.

Methods of Evaluating Genetic Material

There are numerous methods for evaluating genetic material to provide genetic information. These methods can be used to evaluate a genetic locus that includes a gene of the signature set. The methods can be used to evaluate one or more nucleotides, e.g., a coding or non-coding region of the gene, e.g., in a regulatory region (e.g., a promoter, a region encoding an untranslated region or intron, and so forth).

Nucleic acid samples can analyzed using biophysical techniques (e.g., hybridization, electrophoresis, and so forth), sequencing, enzyme-based techniques, and combinations thereof. For example, hybridization of sample nucleic acids to nucleic acid microarrays can be used to evaluate sequences in an mRNA population and to evaluate genetic polymorphisms. Other hybridization based techniques include sequence specific primer binding (e.g., PCR or LCR); Southern analysis of DNA, e.g., genomic DNA; Northern analysis of RNA, e.g., mRNA; fluorescent probe based techniques (see, e.g., Beaudet et al. (2001) Genome Res. 11(4):600-608); and allele specific amplification. Enzymatic techniques include restriction enzyme digestion; sequencing; and single base extension (SBE). These and other techniques are well known to those skilled in the art.

Electrophoretic techniques include capillary electrophoresis and Single-Strand Conformation Polymorphism (SSCP) detection (see, e.g., Myers et al. (1985) Nature 313:495-8 and Ganguly (2002) Hum Mutat. 19(4):334-42). Other biophysical methods include denaturing high pressure liquid chromatography (DHPLC).

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

Enzymatic methods for detecting sequences include amplification based-methods such as the polymerase chain reaction (PCR; Saiki, et al. (1985) Science 230:1350-1354) and ligase chain reaction (LCR; Wu. et al. (1989) Genomics 4:560-569; Barringer et al. (1990), Gene 1989:117-122; F. Barany (1991) Proc. Natl. Acad. Sci. USA 1988:189-193); transcription-based methods utilize RNA synthesis by RNA polymerases to amplify nucleic acid (U.S. Pat. Nos. 6,066,457; 6,132,997; and 5,716,785; Sarkar et al., (1989) Science 244:331-34; Stofler et al., (1988) Science 239:491); NASBA (U.S. Pat. Nos. 5,130,238; 5,409,818; and 5,554,517); rolling circle amplification (RCA; U.S. Pat. Nos. 5,854,033 and 6,143,495) and strand displacement amplification (SDA; U.S. Pat. Nos. 5,455,166 and 5,624,825). Amplification methods can be used in combination with other techniques.

Other enzymatic techniques include sequencing using polymerases, e.g., DNA polymerases and variations thereof such as single base extension technology. See, e.g., U.S. Pat. Nos. 6,294,336; 6,013,431; and 5,952,174.

Fluorescence based detection can also be used to detect nucleic acid polymorphisms. For example, different terminator ddNTPs can be labeled with different fluorescent dyes. A primer can be annealed near or immediately adjacent to a polymorphism, and the nucleotide at the polymorphic site can be detected by the type (e.g., “color”) of the fluorescent dye that is incorporated.

Hybridization to microarrays can also be used to detect polymorphisms, including SNPs. For example, a set of different oligonucleotides, with the polymorphic nucleotide at varying positions with the oligonucleotides can be positioned on a nucleic acid array. The extent of hybridization as a function of position and hybridization to oligonucleotides specific for the other allele can be used to determine whether a particular polymorphism is present. See, e.g., U.S. Pat. No. 6,066,454.

In one implementation, hybridization probes can include one or more additional mismatches to destabilize duplex formation and sensitize the assay. The mismatch may be directly adjacent to the query position, or within 10, 7, 5, 4, 3, or 2 nucleotides of the query position. Hybridization probes can also be selected to have a particular T_m, e.g., between 45-60° C., 55-65° C., or 60-75° C. In a multiplex assay, T_m's can be selected to be within 5, 3, or 2° C. of each other.

It is also possible to directly sequence the nucleic acid for a particular genetic locus (e.g., a gene transcript's locus), e.g., by amplification and sequencing, or amplification, cloning and sequence. High throughput automated (e.g., capillary or microchip based) sequencing apparati can be used. In still other embodiments, the sequence of a protein of interest is analyzed to infer its genetic sequence. Methods of analyzing a protein sequence include protein sequencing, mass spectroscopy, sequence/epitope specific immunoglobulins, and protease digestion.

Kits and Reagents

One or more of the gene transcripts of the transcriptional signature described herein can be used as a component of a kit or as a reagent, e.g., a diagnostic kit or diagnostic reagent. For example, a nucleic acid (or its complement) (e.g., an oligonucleotide, e.g., probe) corresponding to one or more of the genes described herein (or one or more signature sets described herein) can be a member of a nucleic acid array against which a sample (e.g., from a subject, e.g., a subject being evaluated for HCV infection) is hybridized to determine the level of gene expression. For example, a signature set described herein can be present on an array for a TAQMAN® gene expression assay (Applied Biosystems) (e.g., a custom TAQMAN® assay), e.g., for use in a 384-well plate format, e.g., using standard protocols. The diagnostic evaluation of a subject's sample (e.g., peripheral blood) can be performed, e.g., in a doctor's office, hospital laboratory, or contract laboratory.

The nucleic acid can contain the full length gene transcript (or its complement), or a fragment of the transcript (or its complement) (e.g., an oligonucleotide, e.g., probe) that allows for it to specifically bind to the nucleic acid complement (or the nucleic acid) in the sample under selected hybridization conditions. The level can then be compared to a control or reference value. The control or reference value can be part of the kit, or alternatively, the kit can contain the world wide web address on which reference information is located. Alternatively, nucleic acid (or its complement) corresponding to one or more of the genes described herein can be provided as a reagent (e.g., diagnostic reagent) that can be used to detect the presence and level of a gene transcript described herein. For example, the nucleic acid (or its complement) can be labeled with a detectable label and hybridized with nucleic acid from a sample. The level of hybridization can then be compared to a reference value. The reference value can be provided with the reagent, or alternatively, the reagent can contain a world wide web address for a site on which reference information is located.

Likewise, the polypeptide corresponding to a gene described herein can be used as a reagent or as a component of a kit. The polypeptide can be the full length polypeptide or a fragment thereof that allows for it to specifically bind to an antibody or a ligand (e.g., receptor ligand or binding partner or fragment thereof) that is specific for the protein from which the fragment derives, or otherwise allow specific identification of the protein. In another embodiment, antibodies (including intact and/or full length immunoglobulins of types IgA, IgG (e.g., IgG1, IgG2, IgG3, IgG4), IgE, IgD, IgM (as well as subtypes thereof) and antibody fragments, e.g., single chain antibodies, Fab fragments, F(ab′)2 fragments, Fd fragments, Fv fragments, and dAb fragments) specific for one or more polypeptides encoded by gene transcripts can be a reagent or component of a kit for the detection of the polypeptide. For example, a sample can be contacted with the antibody under conditions that allow for binding of the antibody to its antigen and the presence and/or amount of binding are then detected (e.g., by ELISA). Any of the kits can optionally include instructions for its use (e.g., how to use the kit to predict a treatment outcome or to select a treatment regimen, etc.) or can contain a world wide web address to a link where instructions are provided. The reagents may also be supplied with instructions for their use (e.g., how to use the reagents to predict a treatment outcome or to select a treatment regimen, etc.) or a world wide web address to a link where instructions are provided.

As an example, the patterns of expression of a plurality of the genes (e.g., a signature set) described herein in a sample from a subject can be compared with the patterns of expression of the same genes from references, e.g., enhanced responders or non-enhanced responders for a particular therapy (e.g., VX-950 dosing), or non-infected subjects. From the comparison, a prediction can be made, e.g., if the subject's sample has the same or similar pattern of expression of the gene transcripts as the enhanced responder, a prediction can be made that the subject will also respond well to the given therapy. Whether a pattern or expression is the same or similar can be determined by one skilled in the art based upon knowledge of the art, and can optionally include statistical methods.

The kits and reagents can be used, for example, to diagnose HCV, predict the treatment outcome of a subject with HCV (e.g., if the subject is administered a particular therapy), select a treatment regimen (e.g., a monotherapy or combination therapy), select dosages of a given treatment, and/or select the duration of a treatment regimen.

Additional Uses

In one method, information about the subject's gene expression levels, e.g., the result of evaluating a signature set described herein (e.g., a signature set of HCV infection), is provided (e.g., communicated, e.g., electronically communicated) to a third party, e.g., a hospital, clinic, a government entity, reimbursing party or insurance company (e.g., a life insurance company). For example, choice of medical procedure, payment for a medical procedure, payment by a reimbursing party, or cost for a service or insurance can be function of the information. E.g., the third party receives the information, makes a determination based at least in part on the information, and optionally communicates the information or makes a choice of procedure, payment, level of payment, coverage, etc. based on the information.

In one embodiment, a premium for insurance (e.g., life or medical) is evaluated as a function of information about one or more gene expression levels, e.g., a signature set described herein, e.g., a signature set of HCV infection. For example, premiums can be increased (e.g., by a certain percentage) if the genes of a signature set described herein are differentially expressed between an insured candidate (or a candidate seeking insurance coverage) and a reference value (e.g., a non-HCV infected person). As another example, premiums can be decreased if levels of an ISG(s) are sustained (as described herein) after treatment with a viral protease inhibitor (e.g., VX-950) in the an HCV-infected insured candidate or an HCV-infected candidate seeking insurance coverage. Premiums can also be scaled depending on gene expression levels, e.g., the result of evaluating a signature set described herein (e.g., a signature set of HCV infection). For example, premiums can be assessed to distribute risk, e.g., as a function of gene expression levels, e.g., the result of evaluating a signature set described herein (e.g., a signature set of HCV infection). In another example, premiums are assessed as a function of actuarial data that is obtained from subjects that are enhanced or non-enhanced responders.

Information about gene expression levels, e.g., the result of evaluating a signature set described herein (e.g., a signature set of HCV infection), can be used, e.g., in an underwriting process for life insurance. The information can be incorporated into a profile about a subject. Other information in the profile can include, for example, date of birth, gender, marital status, banking information, credit information, children, and so forth. An insurance policy can be recommended as a function of the information on gene expression levels, e.g., the result of evaluating a signature set described herein (e.g., a signature set of HCV infection), along with one or more other items of information in the profile. An insurance premium or risk assessment can also be evaluated as function of the signature set information. In one implementation, points are assigned on the basis of being an enhanced or non-enhanced responder.

In one embodiment, information about gene expression levels, e.g., the result of evaluating a signature set described herein (e.g., a signature set of HCV infection), is analyzed by a function that determines whether to authorize the transfer of funds to pay for a service or treatment provided to a subject (or make another decision referred to herein). For example, the results of analyzing a signature set described herein may indicate that a subject is a non-enhanced responder, suggesting that a longer treatment course is needed, thereby triggering an outcome that indicates or causes authorization to pay for a service or treatment (e.g., a longer duration of anti-HCV therapy, e.g., VX-950 therapy) provided to a subject. For example, an entity, e.g., a hospital, care giver, government entity, or an insurance company or other entity which pays for, or reimburses medical expenses, can use the outcome of a method described herein to determine whether a party, e.g., a party other than the subject patient, will pay for services (e.g., a particular monotherapy or combination therapy, and/or a certain duration of therapy) or treatment provided to the patient. For example, a first entity, e.g., an insurance company, can use the outcome of a method described herein to determine whether to provide financial payment to, or on behalf of, a patient, e.g., whether to reimburse a third party, e.g., a vendor of goods or services, a hospital, physician, or other care-giver, for a service or treatment provided to a patient. For example, a first entity, e.g., an insurance company, can use the outcome of a method described herein to determine whether to continue, discontinue, enroll an individual in an insurance plan or program, e.g., a health insurance or life insurance plan or program.

EXAMPLES

Experiments were performed, in part, to identify a minimal set of gene transcripts associated with chronic HCV infection in clinical samples, establish a baseline gene expression data set (e.g., signature set) in the peripheral blood that may include genes to monitor and correlate with treatment outcomes, and determine if the anti-viral activity of VX-950 results in changes in gene expression in the peripheral blood cells coincident with viral clearance in plasma.

A comparison of baseline peripheral blood samples from healthy and HCV subjects identified a robust, statistically significant set of 258 genes (a signature set) associated with HCV infection (5% false discovery rate). A subset of expression changes in HCV infected patients were of fairly large magnitude (2-fold to 5-fold) and reflected the regulation of genes that have previously been shown to be associated with host antiviral response. Following dosing with VX-950 for 14 days, the expression of these genes tended to normalize towards levels seen in healthy subjects, indicating that VX-950 normalized the signature set, and led to a median 4.4-log drop in HCV plasma viral load (e.g., in subjects dosed with 750 mg VX-950). Sustained levels of interferon-sensitive genes (ISGs) in peripheral blood during VX-950 dosing were associated with an enhanced antiviral response.

Without being bound by theory, it appears that inhibition of NS3/4A by VX-950 may restore IFN signaling, block viral replication in hepatocytes, and block cleavage of TRIF/CARDIF, thereby restoring IRF3 & RIG-1 signaling and transcription of ISGs which activate intrinsic anti-viral defenses, including production of IFNβ, in hepatocytes. Further, it is believed, with respect to plasma clearance of HCV RNA, that B-cells, monocytes, and dendritic cells may take up and degrade HCV particles, and degradation releases viral proteins and dsRNA that activate gene expression in peripheral blood cells. Clearance of plasma HCV RNA and elimination of virus particles can result in normalization of the gene expression signature. In contrast, gene expression persists (e.g., and no normalization occurs) in the presence of 2-3 logs of plasma HCV RNA. Finally, it appears that sustained expression of ISGs in subjects who clear plasma HCV RNA may reflect restored intrinsic antiviral defenses and secretion of interferons. The sustained expression of ISGs may be a sign of the re-emergence of an effective immune response that is essential to eliminate residual HCV infected hepatocytes. Thus, expression of ISGs and other genes associated with acquired immunity may be monitored to establish potential correlations with treatment outcomes.

Example 1
Materials and Methods

The studies presented herein included four panels, each of six healthy subjects, administered placebo, 450 q8h, or 750 q8h, or 1250 mg q12h VX-950 for 5 days and four panels of subjects with HCV administered placebo (six subjects), 450 (ten subjects) q8h, or 750 VX-950 (eight subjects) q8h, or 1250 mg (ten subjects) q12h for 14 days.

RNA Isolation: Peripheral whole blood (2.5 ml) was collected pre-dose and on day-5 from healthy subjects and pre-dose, day-7, -14 and at follow-up from HCV subjects. Total RNA was isolated using standard using PAXGENE BLOOD RNA™ tubes and protocols (Qiagen). Globin transcripts were reduced using the GLOBINCLEA® Human Globin mRNA Removal Kit (Ambion).

Transcriptional Analysis: Transcriptional analyses were performed using Affymetrix U133 v2.0 gene arrays after globin reduction. RNA was prepared using standard protocols and hybridized to Affymetrix Human Genome U133 plus 2.0 arrays.

Data Analysis: Data was processed using Bioconductor, a software, primarily based on R programming language for the analysis and comprehension of genomic data (Bioconductor.org). The data was preprocessed using GCRMA package in Bioconductor, which normalizes at the probe level using the GC content of probes in normalization with RMA (robust multi-array).

Statistically significant differentially expressed genes were identified using SAM algorithm (Significance Analysis of Microarrays) with a false discovery rate of 5%.

Clustering: The statistically significant differentially expressed genes were then subjected to hierarchical (agglomerative) clustering of both genes and subjects using Bioconductor “heatmap” function to identify the minimal set that will distinguish between the two groups.

Example 2
Demographics of HCV Infected Subjects

The study of subjects with chronic HCV infections included six subjects who received a placebo, ten subjects who were dosed with VX-950 at 450 mg q8h, eight subjects who were dosed with VX-950 at 750 mg q8h, and ten subjects who were dosed with VX-950 at 1250 mg q12h. Subject demographics were comparable among groups, except that there were more females in the 750 mg dose group. Only 5 of 28 subjects who received VX-950 had not received prior therapy for HCV. The subject demographics are shown in Table 1.

TABLE 1

Subject Demographics:

450 mg
750 mg
1250 mg

placebo
q8 h
q8 h
q12 h

(n = 6)
(n = 10)
(n = 8)
(n = 10)

Male/female
3/3
8/2
3/5
8/2

Median age (yr)
53
47
52
44

Median wt (kg)
77.2
78.5
75.0
70.0

Treatment-naïve
2
1
1
3

Median HCV RNA (log₁₀)*
6.38
6.45
6.13
6.48

Mean HCV RNA (log₁₀)*
6.28
6.54
6.18
6.46

*HCV RNA levels were determined by the COBAS AmpliPrep/COBAS TAQMAN ™ HCV Test (Roche Molecular Diagnostics).

Example 3
VX-950 Treatment Reduces HCV Viral Loads

The HCV viral loads in HCV infected subjects were examined in each of the groups described in Example 2. As shown in FIG. 1, subjects on placebo had no significant change in viral load (open circles), while all VX-950 dosed subjects had a >2-log initial drop in viral load. All dose groups showed a steep decline of RNA levels in the first 2-3 days. After the initial steep decline over the 3 days, a slower rate of RNA decline was observed in the 750 mg dose group (diamonds), but the median HCV RNA was still decreasing at the end of 14 days. In this assay, for the 450 mg (squares) and 1250 mg (triangles) dose groups, the RNA levels remain more or less stable and even had a tendency to increase again.

Example 4
Signature Set of HCV Infection

Hierarchical clustering analysis revealed a signature set associated with chronic HCV infection. A comparison of genes that are differentially expressed between healthy and HCV-infected subjects at the pre-dose time point revealed a signature set of HCV infection. This signature set consists of 258 genes associated with chronic HCV infection (FDR<5%). The signature set of 258 was identified at baseline, i.e., before the onset of VX-950 dosing. Further, on dosing with VX-950, the expression levels in the HCV-infected patients resolved towards healthy levels, as described in Example 5.

The full list of 258 genes, including the Affymetrix probeset ID, gene symbol, gene description, GO (gene ontology) biological process, GL molecular function, and GL cellular component, is provided in Table 2.

TABLE 2

Genes of an HCV Signature Set

Affymetrix
Gene
Gene
GO Biological

GO Cellular

probeset ID
Symbol
Description
Process
GO Molecular Function
Component

1557961_s_at
—
—
—
—
—

227353_at
—
—
—
—
—

228412_at
—
Full-Length
—
—
—

Cdna Clone

Cs0Df004Yg03

Of Fetal Brain

Of Homo

Sapiens

(Human)

228549_at
—
—
—
—
—

228758_at
—
Hypothetical
—
—
—

Loc389185

232253_at
—
Hypothetical
—
—
—

Gene

Supported By

Ak128882

238768_at
—
Hypothetical
—
—
—

Loc388969

204567_s_at
ABCG1
Atp-Binding
Lipid Transport
Nucleotide Binding ///
Membrane

Cassette, Sub-
/// Cholesterol
Atp Binding /// L-
Fraction ///

Family G
Metabolism ///
Tryptophan
Endoplasmic

(White),
Detection Of
Transporter Activity ///
Reticulum ///

Member 1
Hormone
Purine Nucleotide
Golgi Stack ///

Stimulus ///
Transporter Activity ///
Membrane ///

Response To
Permease Activity ///
Integral To

Organic
Atpase Activity ///
Membrane ///

Substance ///
Atpase Activity,
Integral To

Cholesterol
Coupled To
Plasma

Homeostasis ///
Transmembrane
Membrane

Transport ///
Movement Of

Lipid Transport
Substances /// Protein

/// Transport
Dimerization Activity ///

Atp Binding ///

Nucleoside-

Triphosphatase Activity

/// Atpase Activity,

Coupled To

Transmembrane

Movement Of

Substances /// Atpase

Activity, Coupled To

Transmembrane

Movement Of

Substances

213017_at
ABHD3
Abhydrolase
—
Catalytic Activity ///
—

Domain

Hydrolase Activity

Containing 3

202323_s_at
ACBD3
Acyl-Coenzyme
Steroid
Acyl-Coa Binding ///
Mitochondrion

A Binding
Biosynthesis ///
Protein Carrier Activity
/// Golgi Stack

Domain
Intracellular

/// Membrane

Containing 3
Protein

Transport ///

Lipid

Biosynthesis

201786_s_at
ADAR
Adenosine
Mrna
Dna Binding /// Double-
Nucleus ///

Deaminase,
Processing
Stranded Rna Binding ///
Cytoplasm

Rna-Specific
/// Rna
Double-Stranded Rna
///

Editing ///
Adenosine Deaminase
Intracellular

Antimicrobial
Activity /// Hydrolase Activity
/// Nucleus

Humoral
/// Metal Ion Binding ///

Response
Double-Stranded Rna

(Sensu
Adenosine Deaminase

Vertebrata)
Activity /// Rna Binding ///

/// Base
Double-Stranded Rna

Conversion
Adenosine Deaminase

Or
Activity /// Adenosine

Substitution
Deaminase Activity /// Zinc

Editing ///
Ion Binding /// Double-

Rna
Stranded Rna Adenosine

Processing
Deaminase Activity

239171_at
ADD3
Adducin 3
—
Structural Constituent Of
Cytoskeleton

(Gamma)

Cytoskeleton /// Calmodulin
///

Binding
Membrane

///

Membrane

202912_at
ADM
Adrenomedullin
Camp
Hormone Activity /// Receptor
Extracellular

Biosynthesis
Binding
Space ///

///

Soluble

Progesterone

Fraction ///

Biosynthesis

Extracellular

/// Signal

Region

Transduction

/// Cell-

Cell

Signaling ///

Pregnancy

/// Excretion

///

Circulation

///

Response

To

Wounding

200849_s_at
AHCYL1
S-
One-Carbon
Adenosylhomocysteinase
—

Adenosylhomo
Compound
Activity /// Hydrolase Activity

cysteine
Metabolism

Hydrolase-Like 1

225555_x_at
AKIP
Aurora Kinase
Negative
Protein Binding
Nucleus ///

A Interacting
Regulation

Nucleus

Protein 1
Of Mitosis ///

Positive

Regulation

Of

Proteolysis

222715_s_at
AP1GBP1
Ap1 Gamma
Intracellular
Calcium Ion Binding
Golgi Stack

Subunit Binding
Protein

///

Protein 1
Transport

Membrane

///

/// Ap-1

Endocytosis

Adaptor

///

Complex ///

Transport

Cytoplasm

/// Protein

/// Golgi

Transport

Apparatus

209870_s_at
APBA2
Amyloid Beta
Nervous
Protein Binding /// Protein
—

(A4) Precursor
System
Binding /// Protein Binding

Protein-
Development

Binding, Family
///

A, Member 2
Protein

(X11-Like)
Transport

///

Transport

228520_s_at
APLP2
Amyloid Beta
G-Protein
Dna Binding /// Serine-Type
Nucleus ///

(A4) Precursor-
Coupled
Endopeptidase Inhibitor
Integral To

Like Protein 2
Receptor
Activity /// Protein Binding ///
Membrane

Protein
Dna Binding ///
/// Nucleus

Signaling
Endopeptidase Inhibitor
/// Integral

Pathway
Activity /// Binding
To

Membrane

221653_x_at
APOL2
Apolipoprotein
Lipid
Receptor Binding /// High-
Extracellular

L, 2
Metabolism
Density Lipoprotein Binding
Region ///

/// Lipid
/// Lipid Binding /// Lipid
Intracellular

Transport
Binding

/// Acute-

Phase

Response

///

Development

///

Cholesterol

Metabolism

///

Lipoprotein

Metabolism

///

Transport

225707_at
ARL6IP6
Adp-
—
—
—

Ribosylation-

Like Factor 6

Interacting

Protein 6

209824_s_at
ARNTL
Aryl
Regulation
Transcription Factor Activity ///
Nucleus

Hydrocarbon
Of
Signal Transducer Activity ///

Receptor
Transcription,
Dna Binding /// Transcription

Nuclear
Dna-
Regulator Activity /// Receptor

Translocator-
Dependent
Activity

Like
/// Signal

Transduction

///

Circadian

Rhythm ///

Transcription

///

Regulation

Of

Transcription

208836_at
ATP1B3
Atpase,
Transport
Sodium:Potassium-
Sodium:Potassium

Na+/K+
///
Exchanging Atpase Activity ///
Exchanging

Transporting,
Potassium
Potassium Ion Binding ///
Atpase

Beta 3
Ion
Sodium Ion Binding ///
Complex ///

Polypeptide
Transport
Sodium:Potassium-
Membrane

/// Sodium
Exchanging Atpase Activity
/// Integral

Ion

To

Transport

Membrane

214149_s_at
ATP6V0E
Atpase, H+
Ion
Transporter Activity ///
Membrane

Transporting,
Transport
Hydrolase Activity ///
Fraction ///

Lysosomal
/// Atp
Hydrogen-Transporting Atp
Proton-

9 Kda, V0
Synthesis
Synthase Activity, Rotational
Transporting

Subunit E
Coupled
Mechanism /// Hydrogen-
Two-Sector

Proton
Transporting Atpase Activity,
Atpase

Transport
Rotational Mechanism ///
Complex ///

/// Proton
Hydrogen Ion Transporter
Integral To

Transport
Activity /// Hydrogen-
Membrane

///
Transporting Atpase Activity,

Transport
Rotational Mechanism

/// Proton

Transport

236307_at
BACH2
Btb And Cnc
Transcription
Dna Binding /// Protein
Nucleus

Homology 1,
///
Binding

Basic Leucine
Regulation

Zipper
Of

Transcription
Transcription,

Factor 2
Dna-

Dependent

203140_at
BCL6
B-Cell
Negative Regulation Of
Transcription
Mediator

CII/Lymphoma 6
Transcription From Rna
Factor Activity
Complex ///

(Zinc Finger
Polymerase li Promoter
/// Protein
Nucleus ///

Protein 51) /// B-
/// Transcription ///
Binding /// Zinc
Nucleus

Cell
Regulation Of
Ion Binding ///

CII/Lymphoma 6
Transcription, Dna-
Metal Ion

(Zinc Finger
Dependent ///
Binding ///

Protein 51)
Inflammatory
Nucleic Acid

Response /// Positive
Binding /// Dna

Regulation Of Cell
Binding ///

Proliferation ///
Protein Binding

Regulation Of

Transcription, Dna-

Dependent

228617_at
BIRC4BP
Xiap Associated
—
Zinc Ion Binding
—

Factor-1

243509_at
BTG1
B-Cell
Spermatid
Transcription
Nucleus ///

Translocation
Development ///
Cofactor Activity
Nucleus ///

Gene 1, Anti-
Negative Regulation Of
/// Kinase
Cytoplasm

Proliferative
Cell Proliferation /// Cell
Binding ///

Migration /// Negative
Protein Binding

Regulation Of Cell
/// Enzyme

Growth /// Regulation
Binding

Of Apoptosis ///

Positive Regulation Of

Enzyme Activity ///

Regulation Of

Transcription ///

Positive Regulation Of

Endothelial Cell

Differentiation ///

Positive Regulation Of

Myoblast Differentiation

/// Positive Regulation

Of Angiogenesis

203944_x_at
BTN2A1
Butyrophilin,
Lipid Metabolism
—
Integral To

Subfamily 2,

Membrane

Member A1

/// Integral

To Plasma

Membrane

205298_s_at
BTN2A2
Butyrophilin,
—
—
Integral To

Subfamily 2,

Membrane

Member A2

201457_x_at
BUB3
Bub3 Budding
Mitosis /// Mitotic
—
Kinetochore

Uninhibited By
Spindle Checkpoint ///

/// Nucleus

Benzimidazoles 3
Cell Proliferation ///

Homolog (Yeast)
Mitotic Checkpoint

222464_s_at
C10orf119
Chromosome
—
—
—

10 Open

Reading Frame

119

219471_at
C13orf18
Chromosome
—
Protein Phosphatase
—

13 Open

Inhibitor Activity

Reading Frame

18

222458_s_at
C1orf108
Chromosome 1
—
—
—

Open Reading

Frame 108

212003_at
C1orf144
Chromosome 1
—
—
—

Open Reading

Frame 144

217835_x_at
C20orf24
Chromosome
—
—
—

20 Open

Reading Frame

24

216032_s_at
C20orf47
Chromosome
—
—
Integral To

20 Open

Membrane

Reading Frame

47

223145_s_at
C6orf166
Chromosome 6
—
—
—

Open Reading

Frame 166

243271_at
C7orf6
Sterile Alpha
—
—
—

Motif Domain

Containing

9Like

207181_s_at
CASP7
Caspase 7,
Proteolysis ///
Protein Binding ///
Cytoplasm

Apoptosis-
Apoptotic
Peptidase Activity ///

Related
Program ///
Cysteine-Type

Cysteine
Apoptosis ///
Peptidase Activity ///

Peptidase
Apoptosis
Caspase Activity ///

Cysteine-Type

Peptidase Activity ///

Hydrolase Activity

Rhodopsin-Like
Plasma

Receptor Activity ///
Membrane ///

Receptor Activity ///
Integral To

Protein Binding /// C-C
Plasma

Chemokine Receptor
Membrane ///

Activity /// Signal
Integral To

Transducer Activity ///
Membrane ///

G-Protein Coupled
Plasma

Receptor Activity ///
Membrane

Chemokine Receptor

Activity

205098_at
CCR1
Chemokine (C-C
Chemotaxis ///

Motif) Receptor 1
Inflammatory

Response /// Cell

Adhesion /// G-

Protein Signaling,

Coupled To Cyclic

Nucleotide Second

Messenger ///

Elevation Of

Cytosolic Calcium

Ion Concentration ///

Cell-Cell Signaling

/// Cytokine And

Chemokine

Mediated Signaling

Pathway /// Signal

Transduction ///

GProtein Coupled

Receptor Protein

Signaling Pathway

/// Chemotaxis ///

Immune Response

/// Cell Surface

Receptor Linked

Signal Transduction

/// Response To

Wounding

203547_at
CD4
Cd4 Antigen
Immune Response
Transmembrane
Plasma

(P55) /// Cd4
/// Cell Adhesion ///
Receptor Activity
Membrane ///

Antigen (P55)
Transmembrane
/// Coreceptor
Integral To

Receptor Protein
Activity /// Mhc
Membrane ///

Tyrosine Kinase
Class Ii Protein
T Cell

Signaling Pathway
Binding /// Protein
Receptor

/// T Cell
Binding /// Zinc
Complex ///

Differentiation /// T
Ion Binding ///
Plasma

Cell Selection ///
Receptor Activity
Membrane ///

Positive Regulation
/// Coreceptor
Membrane

Of Interleukin-2
Activity ///

Biosynthesis ///
Receptor Activity

Immune Response

/// Signal

Transduction /// Cell

Surface Receptor

Linked Signal

Transduction ///

Enzyme Linked

Receptor Protein

Signaling Pathway

209287_s_at
CDC42EP3
Cdc42 Effector
Regulation Of
—
Cytoskeleton

Protein (Rho
Cell Shape

Gtpase Binding) 3

212501_at
CEBPB
Ccaat/Enhancer
Transcription
Transcription Factor
Nucleus ///

Binding Protein
/// Regulation
Activity /// Dna
Nucleus

(C/Ebp), Beta
Of
Binding /// Dna

Transcription,
Binding

Dna-

Dependent ///

Transcription

From Rna

Polymerase Ii

Promoter ///

Acute-Phase

Response ///

Inflammatory

Response ///

Immune

Response

205212_s_at
CENTB1
Centaurin, Beta 1
Intracellular
Phospholipase C
—

Signaling
Activity /// Gtpase

Cascade ///
Activator Activity ///

Regulation Of
Metal Ion Binding ///

Gtpase
Zinc Ion Binding

Activity ///

Signal

Transduction

205212_s_at
CENTB1
Centaurin, Beta 1
Intracellular
Phospholipase C
—

Signaling
Activity /// Gtpase

Cascade ///
Activator Activity ///

Regulation
Metal Ion Binding ///

Of Gtpase
Zinc Ion Binding

Activity ///

Signal

Transduction

234562_x_at
CKLFSF8
Chemokine-Like
Chemotaxis
Cytokine Activity
Extracellular

Factor
/// Sensory

Space ///

Superfamily 8
Perception

Membrane ///

Integral To

Membrane

206207_at
CLC
Charcot-Leyden
Phospholipid
Lysophospholipase
—

Crystal Protein
Metabolism
Activity /// Serine

/// Charcot-
///
Esterase Activity ///

Leyden Crystal
Development
Sugar Binding

Protein
/// Lipid
//Hydrolase Activity

Catabolism

///

Antimicrobial

Humoral

Response

(Sensu

Vertebrata)

202160_at
CREBBP
Creb Binding
Response To Hypoxia
Transcription Factor
Nucleus ///

Protein
/// Regulation Of
Activity /// Transcription
Cytoplasm

(Rubinstein-
Transcription, Dna-
Coactivator Activity ///
/// Nucleus

Taybi
Dependent /// Protein
Histone

Syndrome)
Complex Assembly ///
Acetyltransferase

Signal Transduction
Activity /// Signal

/// Homeostasis ///
Transducer Activity ///

Transcription ///
Protein Binding /// Zinc

Regulation Of
Ion Binding ///

Transcription, Dna-
Transferase Activity ///

Dependent ///
Metal Ion Binding ///

Regulation Of
Protein Binding ///

Transcription ///
Transcription Cofactor

Signal Transduction
Activity /// Transcription

/// Regulation Of
Coactivator Activity ///

Transcription
Protein Binding ///

Transcription Coactivator

Activity

212180_at
CRKL
V-Crk
Protein Amino Acid
Protein-Tyrosine Kinase
—

Sarcoma
Phosphorylation ///
Activity /// Sh3/Sh2

Virus Ct10
Cell Motility ///
Adaptor Activity ///

Oncogene
Intracellular Signaling
Protein Binding /// Signal

Homolog
Cascade /// Jnk
Transducer Activity

(Avian)-Like
Cascade /// Ras

Protein Signal

Transduction ///

Intracellular Signaling

Cascade

214743_at
CUTL1
Cut-Like 1,
Negative Regulation
Transcription Factor
Nucleus

Ccaat
Of Transcription From
Activity /// Rna

Displacement
Rna Polymerase Ii
Polymerase Ii

Protein
Promoter ///
Transcription Factor

(Drosophila)
Transcription ///
Activity /// Dna Binding

Development ///

Regulation Of

Transcription, Dna-

Dependent ///

Development ///

Regulation Of

Transcription From

Rna Polymerase Ii

Promoter

214743_at
CUTL1
Cut-Like 1, Ccaat
Negative Regulation
Transcription Factor
Nucleus

Displacement
Of Transcription
Activity /// Rna

Protein
From Rna
Polymerase Ii

(Drosophila)
Polymerase Ii
Transcription Factor

Promoter ///
Activity /// Dna

Transcription ///
Binding

Development ///

Regulation Of

Transcription, Dna-

Dependent ///

Development ///

Regulation Of

Transcription From

Rna Polymerase Ii

Promoter

209164_s_at
CYB561
Cytochrome B-
Electron Transport
Cytochrome-B5
Integral To

561
/// Transport ///
Reductase Activity
Plasma

Generation Of
/// Iron Ion Binding
Membrane ///

Precursor
/// Metal Ion Binding
Integral To

Metabolites And

Membrane

Energy

221903_s_at
CYLD
Cylindromatosis
Ubiquitin-
Cysteine-Type
Cytoskeleton

(Turban Tumor
Dependent Protein
Endopeptidase

Syndrome)
Catabolism ///
Activity /// Ubiquitin

Ubiquitin Cycle ///
Thiolesterase

Cell Cycle ///
Activity /// Ubiquitin

Negative Regulation
Thiolesterase

Of Progression
Activity ///

Through Cell Cycle
Peptidase Activity

/// Ubiquitin-
/// Cysteine-Type

Dependent Protein
Peptidase Activity

Catabolism
/// Hydrolase

Activity

200794_x_at
DAZAP2
Daz Associated
—
—
—

Protein 2

209782_s_at
DBP
D Site Of
Transcription ///
Dna Binding /// Rna
Nucleus

Albumin
Regulation Of
Polymerase Ii

Promoter
Transcription From
Transcription Factor

(Albumin D-Box)
Rna Polymerase Ii
Activity

Binding Protein
Promoter ///

Rhythmic Process

/// Regulation Of

Transcription, Dna-

Dependent

224009_x_at
DHRS9
Dehydrogenase/Reductase
Androgen
Alcohol Dehydrogenase
Microsome

(Sdr Family) Member 9
Metabolism ///
Activity /// Retinol
/// Integral

Progesterone
Dehydrogenase Activity ///
To

Metabolism /// 9-
3-Alpha(17-Beta)-
Endoplasmic

Cis-Retinoic Acid
Hydroxysteroid
Reticulum

Biosynthesis ///
Dehydrogenase (Nad+)
Membrane

Metabolism ///
Activity /// Oxidoreductase
///

Epithelial Cell
Activity /// Racemase And
Membrane

Differentiation ///
Epimerase Activity ///
///

Retinol
Alcohol Dehydrogenase
Microsome

Metabolism ///
Activity /// Retinol
/// Integral

Androgen
Dehydrogenase Activity ///
To

Metabolism ///
3-Alpha(17-Beta)-
Endoplasmic

Epithelial Cell
Hydroxysteroid
Reticulum

Differentiation ///
Dehydrogenase (Nad+)
Membrane

Retinol
Activity

Metabolism /// 9-

Cis-Retinoic Acid

Biosynthesis

208810_at
DNAJB6
Dnaj (Hsp40) Homolog,
Protein Folding ///
Heat Shock Protein
—

Subfamily B, Member 6
Response To
Binding /// Unfolded

Unfolded Protein
Protein Binding

209188_x_at
DR1
Down-Regulator Of
Negative
Dna Binding ///
Nucleus

Transcription 1,
Regulation Of
Transcription Corepressor

TbpBinding (Negative
Transcription
Activity /// Transcription

Cofactor 2)
From Rna
Factor Binding /// Dna

Polymerase Ii
Binding

Promoter ///

Transcription ///

Regulation Of

Transcription,

Dna-Dependent

225415_at
DTX3L
Deltex 3-Like (Drosophila)
Protein
Ubiquitin-Protein Ligase
Ubiquitin

Ubiquitination
Activity /// Zinc Ion Binding
Ligase

/// Metal Ion Binding
Complex

208891_at
DUSP6
Dual Specificity
Regulation Of
Protein Serine/Threonine
Soluble

Phosphatase 6
Progression
Phosphatase Activity ///
Fraction ///

Through Cell
Protein Tyrosine
Cytoplasm

Cycle ///
Phosphatase Activity ///

Inactivation Of
Hydrolase Activity /// Map

Mapk Activity ///
Kinase Phosphatase

Protein Amino
Activity /// Phosphoprotein

Acid
Phosphatase Activity ///

Dephosphorylation
Protein

/// Protein Amino
Tyrosine/Serine/Threonine

Acid
Phosphatase Activity

Dephosphorylation

212830_at
EGFL5
Egf-Like-Domain, Multiple 5
—
Structural Molecule
Integral To

Activity /// Calcium Ion
Membrane

Binding

221497_x_at
EGLN1
Egl Nine
Protein Metabolism
Iron Ion Binding ///
Cytosol

Homolog 1

Oxidoreductase Activity ///

(C. Elegans)

Oxidoreductase Activity,

Acting On Single Donors

With Incorporation Of

Molecular Oxygen,

Incorporation Of Two Atoms

Of Oxygen ///

Oxidoreductase Activity,

Acting On Paired Donors,

With Incorporation Or

Reduction Of Molecular

Oxygen, 2-Oxoglutarate As

One Donor, And

Incorporation Of One

AtomEach Of Oxygen Into

Both Donors /// L-Ascorbic

Acid Binding /// Metal Ion

Binding /// Zinc Ion Binding

214805_at
EIF4A1
Eukaryotic
Protein
Nucleotide Binding /// Dna
—

Translation
Biosynthesis
Binding /// Rna Binding ///

Initiation

Translation Initiation Factor

Factor 4A,

Activity /// Protein Binding ///

Isoform 1

Atp Binding /// Atp-

Dependent Helicase Activity

/// Hydrolase Activity ///

Nucleic Acid Binding ///

Helicase Activity

213579_s_at
EP300
E1A Binding
Response To
Transcription Factor Activity
Nucleus ///

Protein
Hypoxia ///
/// Transcription Coactivator
Nucleus

P300
Regulation Of
Activity /// Histone

Transcription, Dna-
Acetyltransferase Activity ///

Dependent ///
Protein C-Terminus Binding

Apoptosis /// Cell
/// Zinc Ion Binding ///

Cycle /// Signal
Transferase Activity ///

Transduction ///
Metal Ion Binding /// Protein

Nervous System
Binding /// Transcription

Development ///
Factor Binding /// Dna

Homeostasis ///
Binding /// Transcription

Regulation Of
Cofactor Activity ///

Transcription ///
Transcription Coactivator

Transcription ///
Activity /// Protein Binding ///

Regulation Of
Transcription Coactivator

Transcription
Activity

229966_at
EWSR1
Ewing
Transcription ///
Nucleotide Binding /// Rna
Nucleus

Sarcoma
Regulation Of
Binding /// Calmodulin

Breakpoint
Transcription, Dna-
Binding /// Zinc Ion Binding

Region 1
Dependent
/// Metal Ion Binding ///

Nucleic Acid Binding /// Rna

Binding /// Dna Binding ///

Transcription Factor Activity

215206_at
EXT1
Exostoses
Skeletal
Transferase Activity,
Endoplasmic

(Multiple) 1
Development ///
Transferring Glycosyl Groups
Reticulum

Glycosaminoglycan
/// Glucuronosyl-N-
Membrane

Biosynthesis ///
Acetylglucosaminyl-
/// Golgi

Cell Cycle /// Signal
Proteoglycan 4-Alpha-N-
Stack ///

Transduction ///
Acetylglucosaminyltransferase
Membrane

Heparan Sulfate
Activity /// N-
/// Integral

Proteoglycan
Acetylglucosaminyl-
To

Biosynthesis ///
Proteoglycan 4-Beta-
Membrane

Negative
Glucuronosyltransferase
/// Integral

Regulation Of
Activity /// Transferase Activity
To

Progression
/// N-Acetylglucosaminyl-
Endoplasmic

Through Cell Cycle
Proteoglycan 4-Beta-
Reticulum

Glucuronosyltransferase
Membrane

Activity
///

Endoplasmic

Reticulum ///

Integral To

Membrane

///

Endoplasmic

Reticulum ///

Golgi

Apparatus

224840_at
FKBP5
Fk506
Protein Folding ///
Peptidyl-Prolyl Cis-Trans
Nucleus

Binding
ProteinFolding
Isomerase Activity /// Fk506

Protein 5

Binding /// Isomerase Activity

/// Unfolded Protein Binding ///

Protein Binding /// Binding

218999_at
FLJ11000
Hypothetical
—
—
—

Protein

Flj11000

218035_s_at
FLJ20273
Rna-Binding
—
Nucleotide Binding /// Nucleic
—

Protein

Acid Binding /// Rna Binding

219717_at
FLJ20280
Hypothetical
—
—
—

Protein

Flj20280

222751_at
FLJ22313
Hypothetical
Protein
—
—

Protein
Modification

Flj22313

219359_at
FLJ22635
Hypothetical
—
—
—

Protein

Flj22635

230012_at
FLJ34790
Hypothetical
—
—
—

Protein

Flj34790

211074_at
FOLR1
Folate
Receptor Mediated
Receptor Activity /// Folic Acid
Membrane

Receptor 1
Endocytosis ///
Binding /// Receptor Activity ///
Fraction ///

(Adult) ///
Folic Acid
Folic Acid Binding
Integral To

Folate
Transport

Plasma

Receptor 1

Membrane

(Adult)

///

Membrane

209189_at
FOS
V-Fos Fbj
Dna Methylation ///
Dna Binding ///
Nucleus ///

Murine
Regulation Of
Specific Rna
Nucleus

Osteosarcoma
Transcription From
Polymerase Ii

Viral Oncogene
Rna Polymerase Ii
Transcription Factor

Homolog
Promoter ///
Activity

Inflammatory

Response ///

Regulation Of

Transcription, Dna-

Dependent

228188_at
FOSL2
Fos-Like
Regulation Of
Transcription Factor
Nucleus ///

Antigen 2
Transcription From
Activity /// Dna
Nucleus

Rna Polymerase Ii
Binding

Promoter /// Cell

Death /// Regulation

Of Transcription, Dna-

Dependent

200959_at
FUS
Fusion
Immune Response
Nucleotide Binding
Nucleus ///

(Involved In

/// Dna Binding ///
Nucleus ///

T(12; 16) In

Rna Binding ///
Membrane

Malignant

Protein Binding ///

Liposarcoma)

Zinc Ion Binding ///

Metal Ion Binding ///

Nucleic Acid

Binding /// Rna

Binding /// Tumor

Necrosis Factor

Receptor Binding

205483_s_at
G1P2
Interferon,
Protein Modification
Protein Binding
Extracellular

Alpha-Inducible
/// Immune Response

Space ///

Protein (Clone
/// Cell-Cell Signaling

Cytoplasm

Ifi-15K)

204415_at
G1P3
Interferon,
Immune Response ///
—
Integral To

Alpha-Inducible
Response To Pest,

Membrane

Protein (Clone
Pathogen Or Parasite

Ifi-6-16)
/// Immune Response

212804_s_at
GAPVD1
Gtpase
—
—
—

Activating

Protein And

Vps9 Domains 1

209604_s_at
GATA3
Gata Binding
Transcription ///
Transcription Factor
Nucleus

Protein 3
Regulation Of
Activity /// Metal Ion

Transcription, Dna-
Binding /// Dna

Dependent ///
Binding ///

Transcription From
Transcription Factor

Rna Polymerase Ii
Activity /// Zinc Ion

Promoter /// Defense
Binding /// Dna

Response /// Sensory
Binding

Perception Of Sound

/// Morphogenesis

235574_at
GBP4
Guanylate
Immune Response
Gtpase Activity ///
—

Binding Protein 4

Gtp Binding ///

Nucleotide Binding

203925_at
GCLM
Glutamate-
Cysteine Metabolism
Glutamate-Cysteine
—

Cysteine
/// Glutathione
Ligase Activity ///

Ligase,
Biosynthesis
Oxidoreductase

Modifier

Activity /// Ligase

Subunit

Activity

202615_at
GNAQ
Guanine
Protein Amino Acid
Nucleotide Binding
Cytoplasm ///

Nucleotide
Adp-Ribosylation ///
/// Gtpase Activity ///
Heterotrimeric

Binding
Signal Transduction
Signal Transducer
G-Protein

Protein (G
/// G-Protein Coupled
Activity /// Gtp
Complex ///

Protein), Q
Receptor Protein
Binding /// Guanyl
Plasma

Polypeptide
Signaling Pathway ///
Nucleotide Binding
Membrane

Phospholipase C

Activation /// Blood

Coagulation

220404_at
GPR97
G Protein-
Signal Transduction
Receptor Activity ///
Membrane ///

Coupled
/// Neuropeptide
G-Protein Coupled
Integral To

Receptor 97
Signaling Pathway ///
Receptor Activity ///
Membrane ///

G-Protein Coupled
Signal Transducer
Integral To

Receptor Protein
Activity
Membrane

Signaling Pathway

211630_s_at
GSS
Glutathione
Amino Acid
Nucleotide Binding
—

Synthetase ///
Metabolism ///
/// Glutathione

Glutathione
Glutathione
Synthase Activity ///

Synthetase
Biosynthesis ///
Atp Binding ///

Response To
Ligase Activity ///

Oxidative Stress ///
Glutathione

Nervous System
Synthase Activity

Development

204805_s_at
H1FX
H1 Histone
Nucleosome
Dna Binding /// Dna
Nucleosome ///

Family,
Assembly ///
Binding
Nucleus ///

Member X
Chromosome

Chromosome ///

Organization And

Nucleosome

Biogenesis (Sensu

Eukaryota) ///

Nucleosome

Assembly

214500_at
H2AFY
H2A Histone
Nucleosome
Dna Binding /// Dna
Nucleosome ///

Family,
Assembly ///
Binding
Nucleus ///

Member Y
Chromosome

Chromosome ///

Organization And

Barr Body ///

Biogenesis (Sensu

Nucleosome

Eukaryota) /// Dosage

Compensation ///

Nucleosome

Assembly

201007_at
HADHB
Hydroxyacyl-
Lipid Metabolism ///
3-Hydroxyacyl-
Mitochondrial

Coenzyme A
Fatty Acid
Coa
Membrane ///

Dehydrogenase/3-
Metabolism /// Fatty
Dehydrogenase
Mitochondrion

Ketoacyl-
Acid Beta-Oxidation
Activity /// Acetyl-

Coenzyme A
/// Fatty Acid
Coa C-

Thiolase/Enoyl-
Biosynthesis
Acyltransferase

Coenzyme A

Activity /// Enoyl-

Hydratase

Coa Hydratase

(Trifunctional

Activity ///

Protein), Beta

Acyltransferase

Subunit

Activity ///

Transferase

Activity /// Acetyl-

Coa C-

Acyltransferase

Activity ///

Catalytic Activity

217937_s_at
HDAC7A
Histone
Regulation Of
Histone
Histone

Deacetylase 7A
Progression Through
Deacetylase
Deacetylase

Cell Cycle ///
Activity ///
Complex ///

Transcription ///
Transcription
Nucleus ///

Regulation Of
Factor Binding ///
Cytoplasm ///

Transcription, Dna-
Specific
Nucleus

Dependent ///
Transcriptional

Inflammatory
Repressor

Response ///
Activity ///

Nervous System
Hydrolase

Development ///
Activity ///

Chromatin
Protein Binding

Modification /// B Cell

Differentiation ///

Negative Regulation

Of Striated Muscle

Development ///

Chromatin

Modification /// B Cell

Activation

219863_at
HERC5
Hect Domain And
Regulation Of Cyclin
Ubiquitin-Protein
Intracellular

Rld 5
Dependent Protein
Ligase Activity ///

Kinase Activity ///
Ligase Activity

Ubiquitin Cycle ///

ProteinModification

202814_s_at
HEXIM1
Hexamethylene
Negative
Protein Binding ///
Nucleus ///

Bis-Acetamide
Regulation Of
Cyclin-Dependent
Cytoplasm

Inducibl1
Transcription
Protein Kinase

From Rna
Inhibitor Activity

Polymerase Ii
/// Transcriptional

Promoter ///
Repressor

Negative
Activity /// Snrna

Regulation Of
Binding

Cyclin

Dependent

Protein Kinase

Activity

204689_at
HHEX
Hematopoietically
Regulation Of
Transcription
Nucleus /// Nucleus

Expressed
Transcription,
Factor Activity ///

Homeobox
Dna-
Dna Binding ///

Dependent ///
Transcription

Development ///
Factor Activity ///

Antimicrobial
Dna Binding

Humoral

Response

(Sensu

Vertebrata) ///

Development ///

Regulation Of

Transcription

1558561_at
HM13
Histocompatibility
—
Protein Binding ///
Endoplasmic

(Minor) 13

Peptidase Activity
Reticulum ///

/// D-Alanyl-D-
Integral To

Alanine
Membrane

Endopeptidase

Activity ///

Hydrolase Activity

200014_s_at
HNRPC
Heterogeneous
Rna Splicing
Nucleotide
Heterogeneous

Nuclear

Binding /// Rna
Nuclear

Ribonucleoprotein

Binding /// Nucleic
Ribonucleoprotein

C (C1/C2) ///

Acid Binding ///
Complex ///

Heterogeneous

Rna Binding
Nucleus ///

Nuclear

Ribonucleoprotein

Ribonucleoprotein

Complex ///

C (C1/C2)

Nucleus

214918_at
HNRPM
Heterogeneous
—
Nucleotide
Membrane Fraction

Nuclear

Binding /// Rna
/// Nucleus ///

Ribonucleoprotein M

Binding ///
Plasma Membrane

Transmembrane
/// Integral To

Receptor Activity
Plasma Membrane

/// Nucleic Acid
///

Binding ///
Ribonucleoprotein

Receptor Activity
Complex

231271_x_at
HSCARG
Hscarg Protein
Regulation Of
Transcriptional
—

Nitrogen Utilization
Repressor

Activity

202581_at
HSPA1B
Heat Shock 70 Kda
Mrna Catabolism ///
Nucleotide
Nucleus ///

Protein 1B
Protein Folding ///
Binding /// Atp
Cytoplasm

Response To
Binding ///
///

Unfolded Protein ///
Unfolded Protein
Cytoplasm

Protein Biosynthesis
Binding ///

/// Translational
Protein Binding

Elongation ///
/// Translation

Response To
Elongation

Unfolded Protein
Factor Activity ///

Gtp Binding

212493_s_at
HYPB
Huntingtin
—
—
—

Interacting Protein B

202439_s_at
IDS
Iduronate 2-
Metabolism ///
Iduronate-2-
Lysosome

Sulfatase (Hunter
Glycosaminoglycan
Sulfatase Activity
///

Syndrome)
Metabolism
/// Sulfuric Ester
Lysosome

Hydrolase

Activity ///

Hydrolase

Activity ///

Iduronate-2-

Sulfatase Activity

218611_at
IER5
Immediate Early
—
—
—

Response 5

202411_at
IFI27
Interferon, Alpha-
Immune Response ///
—
Integral To

Inducible Protein
Response To Pest,

Membrane

27
Pathogen Or Parasite

/// Integral

To

Membrane

204439_at
IFI44L
Interferon-Induced
—
—
—

Protein 44-Like

203153_at
IFIT1
Interferon-Induced
Immune Response
Binding
Cytoplasm

Protein With

Tetratricopeptide

Repeats 1 ///

InterferoInduced

Protein With

Tetratricopeptide

Repeats 1

217502_at
IFIT2
Interferon-Induced
Immune Response
Binding
—

Protein With

Tetratricopeptide

Repeats 2

229450_at
IFIT3
Interferon-Induced
Immune Response
Binding
—

Protein With

Tetratricopeptide

Repeats 3

203595_s_at
IFIT5
Interferon-Induced
Immune
Binding
—

Protein With
Response

Tetratricopeptide

Repeats 5

201642_at
IFNGR2
Interferon Gamma
Cell Surface
Receptor Activity ///
Integral To

Receptor 2
Receptor Linked
Hematopoietin/Interferon-
Plasma

(Interferon
Signal
Class (D200-Domain)
Membrane

Gamma
Transduction ///
Cytokine Receptor
///

Transducer 1)
Response To
Activity /// Interferon-
Membrane

Virus /// Response
Gamma Receptor Activity
/// Integral

To

To

PathogeniBacteria

Membrane

203126_at
IMPA2
Inositol(Myo)-1(Or
Phosphate
Magnesium Ion Binding
—

4)-
Metabolism ///
/// Inositol-1(Or 4)-

Monophosphatase 2
Signal
Monophosphatase

Transduction
Activity /// Hydrolase

Activity /// Inositol Or

Phosphatidylinositol

Phosphatase Activity ///

Inositol-1(Or 4)-

Monophosphatase

Activity /// Metal Ion

Binding

203275_at
IRF2
Interferon
Negative
Transcription Factor
Nucleus

Regulatory Factor 2
Regulation Of
Activity /// Rna

Transcription
Polymerase Ii

From Rna
Transcription Factor

Polymerase Ii
Activity /// Dna Binding

Promoter ///

Transcription ///

Regulation Of

Transcription,

Dna-Dependent ///

Immune

Response /// Cell

Proliferation

208436_s_at
IRF7
Interferon
Negative Regulation
Transcription Factor
Nucleus ///

Regulatory
Of Transcription
Activity /// Specific
Cytoplasm

Factor 7
From Rna
Rna Polymerase Ii
/// Nucleus

Polymerase Ii
Transcription Factor
/// Nucleus

Promoter ///
Activity /// Dna Binding

Transcription ///
/// Rna Polymerase Ii

Regulation Of
Transcription Factor

Transcription, Dna-
Activity /// Dna Binding

Dependent ///
/// Transcriptional

Transcription
Repressor Activity

Initiation From Rna

Polymerase Ii

Promoter ///

Inflammatory

Response ///

Response To Dna

Damage Stimulus ///

Response To Virus

/// Passive Viral

Induction Of Host

Immune Response ///

Viral Induction Of

Host Immune

Response ///

Response To Virus

/// Negative

Regulation Of

Transcription

203882_at
ISGF3G
Interferon-
Transcription ///
Transcription Factor
Ubiquitin

Stimulated
Regulation Of
Activity /// Ubiquitin-
Ligase

Transcription
Transcription, Dna-
ProteinLigase Activity
Complex ///

Factor 3,
Dependent ///
/// Zinc Ion Binding ///
Nucleus ///

Gamma
Transcription From
Metal Ion Binding ///
Cytoplasm

48 Kda
Rna Polymerase Ii
Dna Binding ///
/// Nucleus

Promoter /// Immune
Transcription Factor

Response /// Cell
Activity

Surface Receptor

Linked Signal

Transduction ///

Response To Virus

/// Protein

Ubiquitination

1553530_a_at
ITGB1
Integrin, Beta
Cellular Defense
Receptor Activity ///
Integrin

1 (Fibronectin
Response /// Cell
Protein Binding ///
Complex ///

Receptor,
Adhesion ///
Protein Binding ///
Integrin

Beta
Homophilic Cell
Protein
Complex ///

Polypeptide,
Adhesion /// Cell-
Heterodimerization
Integral To

Antigen Cd29
Matrix Adhesion ///
Activity /// Protein Self
Membrane

Includes Mdf2,
Integrin-Mediated
Binding

Msk12)
Signaling Pathway ///

Development

209907_s_at
ITSN2
Intersectin 2
Endocytosis
Sh3/Sh2 Adaptor
—

Activity /// Calcium Ion

Binding /// Protein

Binding

223412_at
KBTBD7
Kelch Repeat And
—
Protein Binding
—

Btb (Poz) Domain

Containing 7

227647_at
KCNE3
Potassium
Ion Transport ///
Voltage-Gated
Voltage-

Voltage-Gated
Potassium Ion
Potassium Channel
Gated

Channel, Isk-
Transport ///
Activity /// Potassium
Potassium

Related Family,
Transport
Ion Binding /// Ion
Channel

Member 3

Channel Activity ///
Complex ///

Voltage-Gated Ion
Membrane ///

Channel Activity
Integral To

Membrane

200617_at
KIAA0152
Kiaa0152
—
—
Integral To

Membrane

226808_at
KIAA0543
Likely Ortholog Of
Regulation Of
Nucleic Acid Binding
Intracellular

Mouse Sco-
Transcription,
/// Protein

Spondin
Dna-Dependent
Dimerization Activity

/// Cell Adhesion

229001_at
KIAA1443
Kiaa1443
Regulation Of
Transcription Factor
Nucleus

Transcription,
Activity

Dna-Dependent

233893_s_at
KIAA1530
Kiaa1530 Protein
—
—
—

231956_at
KIAA1618
Kiaa1618
—
Catalytic Activity
—

226720_at
KIAA1935
Kiaa1935 Protein
—
Methyltransferase
—

Activity ///

Transferase Activity

219371_s_at
KLF2
Kruppel-Like
Transcription ///
Transcription Factor
Nucleus ///

Factor 2 (Lung)
Regulation Of
Activity /// Zinc Ion
Nucleus

Transcription,
Binding ///

Dna-Dependent
Transcriptional

Activator Activity ///

Metal Ion Binding ///

Nucleic Acid Binding

/// Dna Binding

1555832_s_at
KLF6
Kruppel-Like
Transcription ///
Dna Binding /// Zinc
Nucleus ///

Factor 6
Regulation Of
Ion Binding ///
Nucleus

Transcription,
Transcriptional

Dna-Dependent
Activator Activity ///

/// B Cell
Metal Ion Binding ///

Differentiation ///
Nucleic Acid Binding

Regulation Of

Transcription,

Dna-Dependent

/// Cell Growth

210313_at
LILRA4
Leukocyte
Immune
Receptor Activity
Integral To

Immunoglobulin-
Response

Membrane

Like Receptor,

Subfamily A (With

Tm Domain),

Member 4

215838_at
LILRA5
Leukocyte
—
—
—

Immunoglobulin-

Like Receptor,

Subfamily A (With

Tm Domain),

Member 5

200704_at
LITAF
Lipopolysaccharide-
Transcription ///
Rna Polymerase
Nucleus

Induced Tnf Factor
Regulation Of
Ii Transcription

Transcription From
Factor Activity ///

Rna Polymerase Ii
Signal

Promoter /// Positive
Transducer

Regulation Of I-
Activity

Kappab Kinase/Nf-

Kappab Cascade ///

Regulation Of

Transcription, Dna-

Dependent

220036_s_at
LMBR1L
Limb Region 1
—
Receptor Activity
—

Homolog (Mouse)-

Like

226375_at
LMTK2
Lemur Tyrosine
Protein Amino Acid
Protein
Integral To

Kinase 2
Phosphorylation ///
Serine/Threonine
Membrane

Protein Amino Acid
Kinase Activity ///
/// Integral

Autophosphorylation
Protein
To

/// Protein Amino
Phosphatase
Membrane

Acid
Inhibitor Activity

Phosphorylation ///
/// Protein

Protein Amino Acid
Binding /// Atp

Phosphorylation ///
Binding ///

Protein Amino Acid
Nucleotide

Autophosphorylation
Binding ///

Protein Kinase

Activity ///

Protein-Tyrosine

Kinase Activity ///

Atp Binding ///

Kinase Activity ///

Transferase

Activity ///

Protein Binding

/// Protein

Serine/Threonine

Kinase Activity ///

Protein

Phosphatase

Inhibitor Activity

/// Atp Binding

226702_at
LOC129607
Hypothetical
Dtdp Biosynthesis ///
Thymidylate
—

Protein Loc129607
DttpBiosynthesis
Kinase Activity ///

Atp Binding ///

Kinase Activity

224990_at
LOC201895
Hypothetical
—
Protein Binding
—

Protein Loc201895

226640_at
LOC221955
Kccr13L
Lipid Metabolism
Triacylglycerol
—

Lipase Activity

225794_s_at
LOC91689
Hypothetical Gene
—
—
—

Supported By

AI449243

228320_x_at
LOC92558
Hypothetical
—
—
—

Protein Loc92558

204692_at
LRCH4
Leucine-Rich
Nervous System
—
—

Repeats And
Development

Calponin Homology

(Ch) Domain

Containing 4

223552_at
LRRC4
Leucine Rich Repeat
—
—
Integral To

Containing 4

Membrane

205859_at
LY86
Lymphocyte Antigen
Apoptosis ///
Signal Transducer
Plasma

86
Inflammatory
Activity
Membrane

Response ///

Humoral Immune

Response ///

Signal

Transduction ///

Cell Proliferation

/// Immune

Response

226748_at
LYSMD2
Lysm, Putative
Cell Wall
—
—

Peptidoglycan-
Catabolism

Binding, Domain

Containing 2

207922_s_at
MAEA
Macrophage
Apoptosis /// Cell
—
Membrane

Erythroblast Attacher
Adhesion ///

Fraction ///

Development

Integral To

Plasma

Membrane

204970_s_at
MAFG
V-Maf
Transcription ///
Transcription
Chromatin

Musculoaponeurotic
Regulation Of
Factor Activity ///
/// Nucleus

Fibrosarcoma
Transcription,
Dna Binding

Oncogene Homolog
Dna-Dependent ///

G (Avian)
Transcription

From Rna

Polymerase Ii

Promoter

228582_x_at
MALAT1
Metastasis
—
—
—

Associated Lung

Adenocarcinoma

Transcript 1 (Non-

Coding Rna)

232333_at
MAML2
Mastermind-Like 2
Transcription ///
Transcription
Nucleus ///

(Drosophila)
Regulation Of
Coactivator
Nucleus

Transcription,
Activity /// Catalytic

Dna-Dependent ///
Activity /// Protein

Notch Signaling
Binding ///

Pathway ///
CampResponse

Positive
Element Binding

Regulation Of
Protein Binding

Transcription

From Rna

Polymerase Ii

Promoter /// Notch

Signaling Pathway

232726_at
MAML3
Mastermind-
Transcription ///
Transcription
Nucleus

Like 3
Regulation Of
Coactivator Activity

(Drosophila)
Transcription,

Dna-Dependent ///

Notch Signaling

Pathway ///

Positive

Regulation Of

Transcription From

Rna Polymerase Ii

Promoter

208785_s_at
MAP1LC3B
Microtubule-
Ubiquitin Cycle ///
Protein Binding
Microtubule

Associated
Autophagy

///

Protein 1

Membrane

Light Chain 3

///

Beta

Autophagic

Vacuole ///

Organelle

Membrane

/// Vacuole

203837_at
MAP3K5
Mitogen-
Mapkkk Cascade
Nucleotide Binding ///
—

Activated
/// Protein Amino
Magnesium Ion

Protein
Acid
Binding /// Protein

Kinase
Phosphorylation ///
Serine/Threonine

Kinase
Apoptosis ///
Kinase Activity /// Map

Kinase 5
Response To
Kinase Kinase Kinase

Stress ///
Activity /// Protein-

Activation Of Jnk
Tyrosine Kinase

Activity ///
Activity /// Atp Binding

Induction Of
/// Transferase Activity

Apoptosis By
/// Protein Self Binding

Extracellular
/// Protein Binding ///

Signals
Protein Kinase Activity

/// Kinase Activity ///

Metal Ion Binding

1552264_a_at
MAPK1
Mitogen-
Protein Amino
Nucleotide Binding ///
—

Activated
Acid
Protein

Protein
Phosphorylation ///
Serine/Threonine

Kinase 1
Induction Of
Kinase Activity /// Map

Apoptosis ///
Kinase Activity ///

Chemotaxis ///
Protein-Tyrosine

Response To
Kinase Activity /// Atp

Stress /// Cell
Binding ///

Cycle /// Signal
Transferase Activity ///

Transduction ///
Protein Kinase Activity

Synaptic
/// Map Kinase Activity

Transmission
/// Kinase Activity

211574_s_at
MCP
Membrane
Immune Response
Receptor Activity
Plasma

Cofactor
/// Complement

Membrane

Protein
Activation,

/// Integral

(Cd46,
Classical Pathway

To Plasma

Trophoblast-
/// Innate Immune

Membrane

Lymphocyte
Response ///

/// Integral

Cross-
Complement

To

Reactive
Activation

Membrane

Antigen)

225742_at
MDM4
Mdm4,
Negative Regulation
Ubiquitin-Protein
Ubiquitin

Transformed
Of Transcription From
Ligase Activity ///
Ligase

3T3 Cell
Rna Polymerase Ii
Protein Binding ///
Complex ///

Double Minute
Promoter /// Protein
Zinc Ion Binding ///
Nucleus ///

4, P53 Binding
Complex Assembly ///
Metal Ion Binding
Nucleus

Protein (Mouse)
Apoptosis /// Cell
/// Zinc Ion Binding

Proliferation ///

Negative Regulation

Of Cell Proliferation

/// Protein

Ubiquitination ///

Negative Regulation

Of Protein

Catabolism /// G0 To

G1 Transition ///

Protein Stabilization

223264_at
MESDC1
Mesoderm
—
—
—

Development

Candidate 1

206522_at
MGAM
Maltase-
Carbohydrate
Glucan 1,4-Alpha-
Integral To

Glucoamylase
Metabolism /// Starch
Glucosidase
Membrane

(Alpha-
Catabolism
Activity ///

Glucosidase)

Hydrolase Activity,

Hydrolyzing O-

Glycosyl

Compounds ///

Alpha-Glucosidase

Activity /// Catalytic

Activity ///

Hydrolase Activity

/// Hydrolase

Activity, Acting On

Glycosyl Bonds ///

Catalytic Activity

225568_at
MGC14141
Hypothetical
—
—
—

Protein

Mgc14141

221756_at
MGC17330
Hgfl Gene ///
—
—
—

Hgfl Gene

244716_x_at
MGC23244
Hypothetical
—
—
—

Protein

Mgc23244

225995_x_at
MGC52000
Cxyorf1-Related
—
—
—

Protein

201298_s_at
MOBK1B
Mob1, Mps One
---
Metal Ion Binding
—

Binder Kinase

/// Zinc Ion

ActivatorLike 1B

Binding

(Yeast)

222555_s_at
MRPL44
Mitochondrial
Rna
Double-Stranded
Mitochondrion ///

Ribosomal
Processing
Rna Binding ///
Ribonucleoprotein

Protein L44

Structural
Complex ///

Constituent Of
Intracellular

Ribosome ///

Endonuclease

Activity ///

Ribonuclease Iii

Activity ///

Hydrolase Activity

/// Rna Binding ///

Nuclease Activity

232724_at
MS4A6A
Membrane-
Signal
Receptor Activity
Integral To

Spanning 4-
Transduction

Membrane

Domains,

Subfamily A,

Member 6A

218773_s_at
MSRB2
Methionine
Protein Repair
Protein-
Mitochondrion

Sulfoxide

Methionine-R-

Reductase B2

Oxide Reductase

Activity ///

Transcription

Factor Activity ///

Zinc Ion Binding

/// Oxidoreductase

Activity

216336_x_at
MT1K
Metallothionein
—
Copper Ion
—

1M

Binding ///

Cadmium Ion

Binding /// Metal

Ion Binding

202086_at
MX1
Myxovirus
Induction Of
Nucleotide
Cytoplasm

(Influenza Virus)
Apoptosis ///
Binding /// Gtpase

Resistance 1,
Immune
Activity /// Gtp

Interferon-
Response ///
Binding /// Gtp

Inducible Protein
Signal
Binding /// Gtpase

P78 (Mouse) ///
Transduction
Activity

Myxovirus
/// Response

(Influenza Virus)
To Virus ///

Resistance 1,
Defense

Interferon-
Response

Inducible Protein

P78 (Mouse)

204994_at
MX2
Myxovirus
Immune Response ///
Nucleotide Binding
Nucleus ///

(Influenza
Response To Virus ///
/// Gtpase Activity ///
Cytoplasm

Virus)
Defense Response
Gtp Binding ///

Resistance

Gtpase Activity

2 (Mouse)

203360_s_at
MYCBP
C-Myc
Transcription ///
Transcription
Nucleus ///

Binding
Regulation Of
Coactivator Activity
Mitochondrion

Protein
Transcription, Dna-
/// Protein Binding
/// Cytoplasm ///

Dependent

Nucleus ///

Cytoplasm

220319_s_at
MYLIP
Myosin
Cell Motility /// Nervous
Ubiquitin-Protein
Ubiquitin

Regulatory
System Development
Ligase Activity ///
Ligase

Light Chain
/// Protein
Cytoskeletal Protein
Complex ///

Interacting
Ubiquitination ///
Binding /// Zinc Ion
Cytoplasm ///

Protein
Ubiquitin Cycle ///
Binding /// Ligase
Cytoskeleton ///

ProteinUbiquitination
Activity /// Metal Ion
Membrane ///

Binding /// Protein
Intracellular

Binding /// Ubiquitin-

Protein Ligase

Activity /// Binding

/// Cytoskeletal

Protein Binding

1567013_at
NFE2L2
Nuclear
Transcription ///
Transcription Factor
Nucleus

Factor
Regulation Of
Activity /// Dna

(Erythroid-
Transcription, Dna-
Binding /// Serine-

Derived 2)-
Dependent ///
Type

Like 2
Transcription From
Endopeptidase

Rna Polymerase Ii
Inhibitor Activity

Promoter

203574_at
NFIL3
Nuclear
Regulation Of
Dna Binding /// Dna
Nucleus ///

Factor,
Transcription, Dna-
Binding ///
Nucleus

Interleukin 3
Dependent ///
Transcription Factor

Regulated
Transcription From
Activity ///

Rna Polymerase Ii
Transcription

Promoter /// Immune
Corepressor Activity

Response

217830_s_at
NSFL1C
Nsfl1 (P97)
—
Lipid Binding
Nucleus ///

Cofactor

Golgi Stack

(P47)

222424_s_at
NUCKS1
Nuclear
—
Kinase Activity
Nucleus

Casein

Kinase And

Cyclin-

Dependent

Kinase

Substrate 1

211973_at
NUDT3
Nudix
Intracellular
Magnesium Ion Binding
Intracellular

(Nucleoside
Signaling
/// Diphosphoinositol-

Diphosphate
Cascade /// Cell-
Polyphosphate

Linked Moiety X)-
Cell Signaling ///
Diphosphatase Activity ///

Type Motif 3
Diadenosine
Hydrolase Activity ///

Polyphosphate
Diphosphoinositol-

Catabolism ///
Polyphosphate

Calcium-
Diphosphatase Activity ///

Mediated
Metal Ion Binding ///

Signaling ///
Diphosphoinositol-

Cyclic-
Polyphosphate

Nucleotide-
Diphosphatase Activity

Mediated

Signaling ///

Regulation Of

Rna Export From

Nucleus ///

Intracellular

Transport

204972_at
OAS2
2′-5′-
Nucleobase,
Rna Binding /// Atp
Microsome ///

Oligoadenylate
Nucleoside,
Binding/// Transferase
Membrane

Synthetase 2,
Nucleotide And
Activity ///

69/71 Kda
Nucleic Acid
Nucleotidyltransferase

Metabolism ///
Activity /// Nucleic Acid

Immune
Binding

Response

218400_at
OAS3
2′-5′-
Nucleobase,
Rna Binding /// Atp
Microsome

Oligoadenylate
Nucleoside,
Binding/// Transferase

Synthetase 3,
Nucleotide And
Activity ///

100 Kda
Nucleic Acid
Nucleotidyltransferase

Metabolism ///
Activity /// Nucleic Acid

Immune
Binding

Response

205660_at
OASL
2′-5′-
Protein
Dna Binding /// Double-
Nucleolus ///

Oligoadenylate
Modification ///
Stranded Rna Binding ///
Cytoplasm

Synthetase-Like
Immune
Atp Binding ///

Response
Transferase Activity ///

Thyroid Hormone

Receptor Binding ///

Nucleic Acid Binding ///

Rna Binding

201599_at
OAT
Ornithine
Amino Acid
Ornithine-Oxo-Acid
Mitochondrial

Aminotransferase
Metabolism ///
Transaminase Activity ///
Matrix ///

(Gyrate Atrophy)
Ornithine
Transferase Activity ///
Mitochondrion

Metabolism ///
Pyridoxal Phosphate
///

Visual Perception
Binding /// Ornithine-
Mitochondrion

Oxo-Acid Transaminase

Activity /// Transaminase

Activity

205760_s_at
OGG1
8-Oxoguanine
Carbohydrate
Damaged Dna Binding ///
Nucleoplasm

Dna
Metabolism ///
Endonuclease Activity ///
///

Glycosylase
Base-Excision
Purine-Specific Oxidized
Mitochondrion

Repair /// Dna
Base Lesion Dna N-
/// Nucleus

Repair /// Base-
Glycosylase Activity ///

Excision Repair
Hydrolase Activity, Acting

/// Response To
On Glycosyl Bonds ///

Dna Damage
Lyase Activity /// Dna

Stimulus /// Dna
Binding /// Catalytic

Repair
Activity /// Dna-(Apurinic

Or Apyrimidinic Site)

Lyase Activity /// Purine-

Specific Oxidized Base

Lesion Dna N-Glycosylase

Activity /// Hydrolase

Activity /// Purine-Specific

Oxidized Base Lesion Dna

N-Glycosylase Activity

207091_at
P2RX7
Purinergic
Ion Transport ///
Receptor Activity /// Atp-
Integral To

Receptor P2X,
Signal
Gated Cation Channel
Plasma

Ligand-Gated
Transduction ///
Activity /// Ion Channel
Membrane ///

Ion Channel, 7
Transport ///
Activity /// Atp Binding ///
Membrane ///

Transport
Receptor Activity
Integral To

Membrane

218809_at
PANK2
Pantothenate
Coenzyme A
Nucleotide Binding ///
—

Kinase 2
Biosynthesis
Pantothenate Kinase

(Hallervorden-

Activity /// Atp Binding ///

Spatz

Transferase Activity ///

Syndrome)

Kinase Activity

223220_s_at
PARP9
Poly (Adp-
Protein Amino
Nad+ Adp-
Nucleus ///

Ribose)
Acid Adp-
Ribosyltransferase Activity
Nucleus

Polymerase
Ribosylation ///

Family,
Cell Migration

Member 9

203708_at
PDE4B
Phosphodiesterase
Signal
Camp-Specific
Soluble

4B, Camp-Specific
Transduction
Phosphodiesterase
Fraction ///

(Phosphodiesterase

Activity /// Hydrolase
Insoluble

E4 Dunce Homolog,

Activity /// Catalytic
Fraction

Drosophila)

Activity /// 3′,5′-Cyclic-

Nucleotide

Phosphodiesterase

Activity

207668_x_at
PDIA6
Protein Disulfide
Electron
Protein Disulfide
Endoplasmic

Isomerase Family A,
Transport ///
Isomerase Activity ///
Reticulum

Member 6
Protein Folding
Electron Transporter

Activity /// Isomerase

Activity /// Protein

Disulfide Isomerase

Activity

202464_s_at
PFKFB3
6-Phosphofructo-2-
Fructose 2,6-
Nucleotide Binding ///
—

Kinase/Fructose-2,6-
Bisphosphate
Catalytic Activity /// 6-

Biphosphatase 3
Metabolism ///
Phosphofructo-2-

Fructose 2,6-
Kinase Activity ///

Bisphosphate
Fructose-2,6-

Metabolism ///
Bisphosphate 2-

Metabolism
Phosphatase Activity

/// Atp Binding ///

Kinase Activity ///

Transferase Activity ///

Hydrolase Activity ///

6-Phosphofructo2-

Kinase Activity

218517_at
PHF17
Phd Finger Protein
Regulation Of
Protein Binding ///
Nucleus ///

17
Transcription,
Zinc Ion Binding ///
Cytoplasm ///

Dna-Dependent
Protein Binding ///
Nucleus ///

/// Apoptosis ///
Protein Binding
Cytoplasm

Response To

Stress ///

Negative

Regulation Of

Cell Growth ///

Apoptosis ///

Response To

Stress ///

Negative

Regulation Of

Cell Growth

203278_s_at
PHF21A
Phd Finger Protein
Regulation Of
Protein Binding ///
—

21A
Transcription,
Zinc Ion Binding ///

Dna-Dependent
Dna Binding ///

/// Transcription
Helicase Activity ///

Metal Ion Binding

203691_at
PI3
Peptidase
Copulation
Serine-Type
Extracellular

Inhibitor 3,

Endopeptidase
Matrix (Sensu

Skin-Derived

Inhibitor Activity ///
Metazoa) ///

(Skalp) ///

Protein Binding ///
Extracellular

Peptidase

Endopeptidase
Region

Inhibitor 3,

Inhibitor Activity ///

Skin-Derived

Serine-Type

(Skalp)

Endopeptidase

Inhibitor Activity ///

Endopeptidase

Inhibitor Activity

210845_s_at
PLAUR
Plasminogen
Cell Motility ///
Protein Binding /// U-
Plasma

Activator,
Chemotaxis /// Cell
Plasminogen
Membrane ///

Urokinase
Surface Receptor
Activator Receptor
Cell Surface ///

Receptor
Linked Signal
Activity /// Receptor
Integral To

Transduction ///
Activity /// U-
Membrane ///

Blood Coagulation
Plasminogen
Extrinsic To

/// Regulation Of
Activator Receptor
Membrane ///

Proteolysis ///
Activity /// Receptor
Membrane

Signal
Activity /// Receptor

Transduction ///
Activity /// Kinase

Blood Coagulation
Activity

202430_s_at
PLSCR1
Phospholipid
Response To Virus
Calcium Ion Binding
Plasma

Scramblase 1
/// Phospholipid
/// Phospholipid
Membrane ///

Scrambling ///
Scramblase Activity
Integral To

Platelet Activation
/// Calcium Ion
Membrane

Binding

200695_at
PPP2R1A
Protein
Regulation Of
Antigen Binding ///
Protein

Phosphatase
Progression Through
Phosphoprotein
Phosphatase

2 (Formerly
Cell Cycle ///
Phosphatase
Type 2A

2A),
Inactivation Of Mapk
Activity /// Protein
Complex ///

Regulatory
Activity /// Regulation
Binding /// Protein
Soluble

Subunit A (Pr
Of Dna Replication ///
Phosphatase Type
Fraction ///

65), Alpha
Regulation Of
2A Regulator
Nucleus ///

Isoform
Translation /// Protein
Activity /// Hydrolase
Mitochondrion

Complex Assembly ///
Activity /// Protein
/// Cytosol ///

Protein Amino Acid
Heterodimerization
Microtubule

Dephosphorylation ///
Activity /// Binding
Cytoskeleton

Ceramide Metabolism

/// Membrane

/// Induction Of

Apoptosis /// Rna

Splicing /// Response

To Organic

Substance ///

Second-Messenger-

Mediated Signaling ///

Regulation Of Wnt

Receptor Signaling

Pathway ///

Regulation Of Cell

Adhesion /// Negative

Regulation Of Cell

Growth /// Regulation

Of Growth ///

Negative Regulation

Of Tyrosine

Phosphorylation Of

Stat3 Protein ///

Regulation Of

Transcription ///

Regulation Of Cell

Differentiation

201859_at
PRG1
Proteoglycan
—
—
—

1, Secretory

Granule

201762_s_at
PSME2
Proteasome
Immune Response
Proteasome
Proteasome

(Prosome,

Activator Activity
Complex

Macropain)

(Sensu

Activator

Eukaryota) ///

Subunit 2

Proteasome

(Pa28 Beta)

Activator

Complex ///

Cytosol ///

Protein

Complex

201433_s_at
PTDSS1
Phosphatidylserine
Phosphatidylserine
Transferase
Integral To

Synthase 1
Biosynthesis ///
Activity
Membrane

Phospholipid

Biosynthesis

200730_s_at
PTP4A1
Protein Tyrosine
Protein Amino Acid
Protein Tyrosine
Endoplasmic

Phosphatase Type
Dephosphorylation
Phosphatase
Reticulum ///

Iva, Member 1
/// Cell Cycle ///
Activity ///
Membrane

Development
Hydrolase Activity

/// Phosphoprotein

Phosphatase

Activity

208616_s_at
PTP4A2
Protein Tyrosine
Protein Amino Acid
Prenylated Protein
Membrane

Phosphatase Type
Dephosphorylation
Tyrosine

Iva, Member 2

Phosphatase

Activity ///

Hydrolase Activity

/// Phosphoprotein

Phosphatase

Activity /// Protein

Tyrosine

Phosphatase

Activity

205174_s_at
QPCT
Glutaminyl-Peptide
Protein Modification
Peptidase Activity
—

Cyclotransferase
/// Proteolysis
/// Acyltransferase

(Glutaminyl

Activity ///

Cyclase)

Glutaminyl-

Peptide

Cyclotransferase

Activity ///

Transferase

Activity

209514_s_at
RAB27A
Rab27A, Member
Intracellular Protein
Nucleotide Binding
—

Ras Oncogene
Transport /// Small
/// Gtpase Activity

Family
Gtpase Mediated
/// Gtp Binding

Signal Transduction

/// Protein Transport

221808_at
RAB9A
Rab9A, Member
Intracellular Protein
Nucleotide Binding
Golgi Stack

Ras Oncogene
Transport /// Small
/// Gtpase Activity
/// Lysosome

Family
Gtpase Mediated
/// Gtp Binding
/// Late

Signal Transduction

Endosome

/// Transport ///

Protein Transport

202100_at
RALB
V-Ral Simian
Intracellular Protein
Nucleotide Binding
—

Leukemia Viral
Transport /// Signal
/// Gtp Binding ///

Oncogene
Transduction ///
Gtp Binding

Homolog B (Ras
Small Gtpase

Related; Gtp
Mediated Signal

Binding Protein)
Transduction

244674_at
RBM6
Rna Binding Motif
Rna Processing
Nucleotide Binding
Nucleus ///

Protein 6

/// Dna Binding ///
Intracellular ///

Rna Binding ///
Nucleus

Nucleic Acid Binding

/// Rna Binding

217775_s_at
RDH11
Retinol
Metabolism ///
Retinol
Intracellular ///

Dehydrogenase
Retinol
Dehydrogenase
Endoplasmic

11 (All-Trans And
Metabolism ///
Activity ///
Reticulum ///

9-Cis)
Photoreceptor
Oxidoreductase
Integral To

Maintenance ///
Activity
Membrane

Visual

Perception

229285_at
RNASEL
Ribonuclease L
Mrna
Rna Binding ///
—

(2′,5′-
Processing ///
Protein

Oligoisoadenylate
Protein Amino
Serine/Threonine

Synthetase-
Acid
Kinase Activity ///

Dependent)
Phosphorylation
Atp Binding ///

/// Protein
Hydrolase Activity ///

Amino Acid
Endoribonuclease

Phosphorylation
Activity, Producing

5′-

Phosphomonoesters

/// Metal Ion Binding

/// Nucleotide

Binding /// Protein

Kinase Activity ///

Kinase Activity ///

Transferase Activity

225414_at
RNF149
Ring Finger
Proteolysis ///
Ubiquitin-Protein
Ubiquitin Ligase

Protein 149
Protein
Ligase Activity ///
Complex

Ubiquitination
Peptidase Activity ///

Zinc Ion Binding

224947_at
RNF26
Ring Finger
Protein
Ubiquitin-Protein
Ubiquitin Ligase

Protein 26
Ubiquitination
Ligase Activity ///
Complex ///

Zinc Ion Binding ///
Nucleus

Metal Ion Binding ///

Zinc Ion Binding

219035_s_at
RNF34
Ring Finger
Apoptosis ///
Ubiquitin-Protein
Ubiquitin Ligase

Protein 34
Protein
Ligase Activity ///
Complex ///

Ubiquitination ///
Zinc Ion Binding ///
Nucleus ///

Ubiquitin Cycle
Metal Ion Binding
Membrane

211976_at
RPL35
Ribosomal
Protein
Mrna Binding ///
Nucleolus ///

Protein L35
Biosynthesis ///
Structural
Ribosome ///

Protein
Constituent Of
Cytosolic Large

Biosynthesis
Ribosome ///
Ribosomal

Structural
Subunit (Sensu

Constituent Of
Eukaryota) ///

Ribosome
Intracellular ///

Ribonucleoprotein

Complex

213797_at
RSAD2
Radical S-
—
Catalytic
—

Adenosyl

Activity ///

Methionine

Iron Ion

Domain

Binding

Containing 2

210968_s_at
RTN4
Reticulon 4
Negative
Protein
Nuclear Membrane ///

Regulation Of Anti-
Binding
Endoplasmic

Apoptosis ///

Reticulum /// Integral

Negative

To Membrane ///

Regulation Of Axon

Integral To

Extension ///

Endoplasmic

Regulation Of

Reticulum Membrane

Apoptosis ///

/// Endoplasmic

Apoptosis

Reticulum

222986_s_at
SCOTIN
Scotin
Positive Regulation
Signal
Nucleus

Of I-Kappab
Transducer

Kinase/Nf-Kappab
Activity

Cascade

202228_s_at
SDFR1
Stromal Cell
—
Receptor
Membrane

Derived

Activity

Factor

Receptor 1

209206_at
SEC22L1
Sec22 Vesicle
Er To Golgi
—
Endoplasmic

Trafficking
Transport /// Protein

Reticulum Membrane

Protein-Like 1
Transport ///

/// Golgi Stack ///

(S. Cerevisiae)
Vesicle-Mediated

Integral To Membrane

Transport ///

/// Endoplasmic

Transport /// Er To

Reticulum

Golgi Transport

201582_at
SEC23B
Sec23
Intracellular Protein
Protein
Endoplasmic

Homolog B (S. Cerevisiae)
Transport /// Er To
Binding
Reticulum /// Golgi

Golgi Transport ///

Stack /// Membrane ///

Vesicle-Mediated

Copii Vesicle Coat

Transport ///

Transport /// Protein

Transport

212268_at
SERPINB1
Serpin Peptidase Inhibitor,
—
Serine-Type
Cytoplasm

Clade B (Ovalbumin),

Endopeptidase

Member 1

Inhibitor

Activity ///

Endopeptidase

Inhibitor

Activity ///

Serine-Type

Endopeptidase

Inhibitor

Activity

208313_s_at
SF1
Splicing Factor 1
Spliceosome
Rna
Spliceosome

Assembly ///
Polymerase Ii
Complex ///

Transcription
Transcription
Ribosome ///

/// Regulation
Factor Activity
Nucleus ///

Of
///
Nucleus

Transcription,
Transcription

Dna-
Corepressor

Dependent ///
Activity /// Rna

Nuclear Mrna
Binding ///

Splicing, Via
Metal Ion

Spliceosome
Binding ///

/// Mrna
Nucleic Acid

Processing
Binding /// Rna

Binding /// Zinc

Ion Binding ///

Nucleic Acid

Binding ///

Metal Ion

Binding

225056_at
SIPA1L2
Signal-Induced
—
Gtpase
—

Proliferation-Associated 1

Activator

Like 2

Activity ///

Protein

Binding

203761_at
SLA
Src-Like-Adaptor /// Src-
Intracellular
Sh3/Sh2
—

Like-Adaptor
Signaling
Adaptor

Cascade
Activity

205896_at
SLC22A4
Solute Carrier Family 22
Ion Transport
Nucleotide
Plasma

(Organic Cation
/// Sodium
Binding /// Atp
Membrane

Transporter), Member 4
Ion Transport
Binding ///
/// Integral

/// Fluid
Organic Cation
To Plasma

Secretion ///
Porter Activity
Membrane

Organic
/// Ion
///

Cation
Transporter
Membrane

Transport ///
Activity ///
/// Integral

Transport
Symporter
To

Activity ///
Membrane

Sodium Ion

Binding ///

Nucleotide

Binding ///

Transporter

Activity

218749_s_at
SLC24A6
Solute Carrier Family 24
—
—
Integral To

(Sodium/Potassium/Calcium

Membrane

Exchanger), Member 6

202497_x_at
SLC2A3
Solute Carrier
Carbohydrate
Transporter Activity ///
Membrane

Family 2
Metabolism ///
Sugar Porter Activity ///
Fraction ///

(Facilitated
Carbohydrate
Glucose Transporter
Membrane ///

Glucose
Transport ///
Activity /// Glucose
Integral To

Transporter),
Glucose
Transporter Activity
Membrane ///

Member 3
Transport ///

Integral To

Transport ///

Membrane

Development ///

Spermatogenesis

/// Cell

Differentiation

235013_at
SLC31A1
Solute Carrier
Ion Transport ///
Copper Ion Transporter
Integral To

Family 31
Copper Ion
Activity /// Copper Ion
Plasma

(Copper
Transport ///
Transporter Activity ///
Membrane ///

Transporters),
Copper Ion
Copper Ion Binding
Integral To

Member 1
Transport ///

Membrane

Transport

225175_s_at
SLC44A2
Solute Carrier
Transport ///
Signal Transducer
Integral To

Family 44,
Positive
Activity
Membrane

Member 2
Regulation Of I-

Kappab

Kinase/Nf-

Kappab Cascade

209131_s_at
SNAP23
Synaptosomal-
Transport ///
T-Snare Activity
Membrane ///

Associated
Protein Transport

Synaptosome ///

Protein, 23 Kda
/// Post-Golgi

Plasma

Transport ///

Membrane

Vesicle Targeting

/// Membrane

Fusion

208821_at
SNRPB
Small Nuclear
Mrna Processing
Rna Binding /// Protein
Spliceosome

Ribonucleoprotein
/// Rna Splicing
Binding
Complex /// Small

Polypeptides B
/// Nuclear Mrna

Nucleolar

And B1
Splicing, Via

Ribonucleoprotein

Spliceosome

Complex /// Small

Nuclear

Ribonucleoprotein

Complex ///

Nucleus ///

Ribonucleoprotein

Complex /// Small

Nucleolar

Ribonucleoprotein

Complex

221561_at
SOAT1
Sterol O-
Lipid Metabolism
Sterol O-Acyltransferase
Endoplasmic

Acyltransferase
/// Circulation ///
Activity ///
Reticulum ///

(Acyl-Coenzyme
Steroid
Acyltransferase Activity
Membrane ///

A: Cholesterol
Metabolism ///
/// Acyltransferase
Integral To

Acyltransferase) 1
Cholesterol
Activity /// Transferase
Membrane ///

Metabolism ///
Activity
Endoplasmic

Cholesterol

Reticulum

Metabolism

208012_x_at
SP110
Sp110 Nuclear
Transcription ///
Dna Binding ///
Nucleus ///

Body Protein
Regulation Of
Hematopoietin/Interferon-
Nucleus

Transcription,
Class (D200-Domain)

Dna-Dependent
Cytokine Receptor Signal

/// Electron
Transducer Activity ///

Transport
Protein Binding /// Zinc

Ion Binding /// Metal Ion

Binding /// Dna Binding ///

Electron Transporter

Activity

221769_at
SPSB3
Spla/Ryanodine
Intracellular
—
—

Receptor
Signaling Cascade

Domain And

Socs Box

Containing 3

217995_at
SQRDL
Sulfide Quinone
—
Oxidoreductase
Mitochondrion

Reductase-Like

Activity

(Yeast)

201247_at
SREBF2
Sterol
Regulation Of
Dna Binding ///
Nucleus ///

Regulatory
Transcription From
Rna Polymerase Ii
Endoplasmic

Element Binding
Rna Polymerase Ii
Transcription
Reticulum ///

Transcription
Promoter /// Lipid
Factor Activity ///
Golgi Stack ///

Factor 2
Metabolism ///
Protein Binding ///
Integral To

Steroid Metabolism
Transcription
Membrane

/// Cholesterol
Regulator Activity

Metabolism ///

Transcription ///

Regulation Of

Transcription, Dna-

Dependent /// Lipid

Metabolism ///

Regulation Of

Transcription

208921_s_at
SRI
Sorcin
Regulation Of
Receptor Binding
Cytoplasm

Action Potential ///
/// Calcium

Transport ///
Channel

Intracellular
Regulator Activity

Sequestering Of
/// Calcium Ion

Iron Ion ///
Binding

Regulation Of

Striated Muscle

Contraction /// Heart

Development ///

Muscle

Development ///

Regulation Of Heart

Contraction Rate

210190_at
STX11
Syntaxin 11
Intracellular Protein
Snap Receptor
Golgi Stack ///

Transport ///
Activity /// Protein
Membrane

Membrane Fusion
Transporter

/// Transport ///
Activity

Protein Transport

208831_x_at
SUPT6H
Suppressor Of Ty
Nucleobase,
Transcription Factor
Nucleus ///

6 Homolog (S. Cerevisiae)
Nucleoside,
Activity /// Rna
Nucleus

Nucleotide And
Binding /// Hydrolase

Nucleic Acid
Activity, Acting On

Metabolism ///
Ester Bonds

Chromatin

Remodeling ///

Regulation Of

Transcription,

Dna-

Dependent ///

Intracellular

Signaling

Cascade ///

Transcription

/// Regulation

Of

Transcription,

Dna-

Dependent

229723_at
TAGAP
T-Cell Activation
—
Guanyl-Nucleotide
—

Gtpase Activating

Exchange Factor

Protein

Activity

202307_s_at
TAP1
Transporter 1,
Transport ///
Nucleotide Binding
Endoplasmic

Atp-Binding
Oligopeptide
/// Transporter
Reticulum ///

Cassette, Sub-
Transport ///
Activity /// Atp
Integral To

Family B
Immune
Binding ///
Membrane ///

(Mdr/Tap)
Response ///
Oligopeptide
Integral To

Protein
Transporter Activity
Membrane

Transport ///
/// Atpase Activity ///

Peptide
Atpase Activity,

Transport
Coupled To

Transmembrane

Movement Of

Substances ///

Protein

Heterodimerization

Activity ///

Nucleoside-

Triphosphatase

Activity

201174_s_at
TERF2IP
Telomeric
Telomerase-
Telomeric Dna
Nuclear

Repeat Binding
Dependent
Binding /// Dna
Chromosome

Factor 2,
Telomere
Binding /// Receptor
///

Interacting
Maintenance ///
Activity
Chromosome,

Protein
Regulation Of

Telomeric

Transcription ///

Region ///

Telomere

Nucleus ///

Maintenance ///

Chromosome

Transcription ///

Regulation Of

Transcription,

Dna-Dependent

205016_at
TGFA
Transforming
Regulation Of
Protein-Tyrosine
Extracellular

Growth Factor,
Progression
Kinase Activity ///
Space ///

Alpha
Through Cell
Signal Transducer
Soluble

Cycle /// Cell-Cell
Activity /// Epidermal
Fraction ///

Signaling /// Cell
Growth Factor
Plasma

Proliferation ///
Receptor Activating
Membrane ///

Cell Proliferation
Ligand Activity ///
Integral To

Protein Binding ///
Plasma

Growth Factor
Membrane ///

Activity
Integral To

Membrane

230651_at
THOC2
Tho Complex 2
Nuclear Mrna
Rna Binding
Nucleus

Splicing, Via

Spliceosome ///

Mrna Export From

Nucleus ///

Transport /// Mrna

Processing

242617_at
TMED8
Transmembrane
Intracellular
Protein Carrier
Membrane

Emp24 Protein
Protein Transport
Activity

Transport

Domain

Containing 8

217795_s_at
TMEM43
Transmembrane
—
—
Integral To

Protein 43

Membrane

200620_at
TMEM59
Transmembrane
—
—
Integral To

Protein 59

Membrane

203839_s_at
TNK2
Tyrosine
Protein Amino
Nucleotide Binding ///
Cytoplasm

Kinase, Non-
Acid
Protein

Receptor, 2
Phosphorylation
Serine/Threonine

/// Cytoskeleton
Kinase Activity ///

Organization And
Non-Membrane

Biogenesis ///
Spanning Protein

Small Gtpase
Tyrosine Kinase

Mediated Signal
Activity /// Gtpase

Transduction
Inhibitor Activity ///

Protein Binding ///

Atp Binding ///

Transferase Activity

/// Protein Kinase

Activity /// Protein-

Tyrosine Kinase

Activity /// Kinase

Activity

221507_at
TNPO2
Transportin 2
Protein Import Into
Binding /// Nuclear
Nucleus ///

(Importin 3,
Nucleus, Docking ///
Localization
Nuclear Pore

Karyopherin
Protein Transport ///
Sequence Binding ///
/// Cytoplasm

Beta 2B)
Transport
Protein Transporter
/// Nucleus ///

Activity
Cytoplasm

237895_at
TNRC6B
Trinucleotide
Intracellular Protein
Nucleotide Binding ///
—

Repeat
Transport /// Small
Gtp Binding

Containing 6B
Gtpase Mediated

Signal Transduction

/// Protein Transport

217914_at
TPCN1
Two Pore
Transport /// Ion
Ion Channel Activity ///
Membrane ///

Segment
Transport /// Cation
Cation Channel
Integral To

Channel 1
Transport
Activity /// Calcium Ion
Membrane

Binding

221571_at
TRAF3
Tnf Receptor-
Induction Of
Ubiquitin-Protein
Ubiquitin

Associated
Apoptosis /// Signal
Ligase Activity ///
Ligase

Factor 3
Transduction ///
Signal Transducer
Complex

Protein
Activity /// Protein

Ubiquitination ///
Binding /// Zinc Ion

Regulation Of
Binding /// Metal Ion

Apoptosis ///
Binding /// Receptor

Apoptosis /// Signal
Activity

Transduction

216749_at
TRERF1
Transcriptional
Steroid
Transcription Factor
Nucleus ///

Regulating
Biosynthesis ///
Activity ///
Nucleus

Factor 1
Cholesterol
Transcription Factor

Catabolism ///
Binding /// Zinc Ion

Development ///
Binding /// Dna

Homeostasis ///
Bending Activity ///

Regulation Of
Rna Polymerase Ii

Transcription ///
Transcription Mediator

Positive Regulation
Activity /// Ligand-

Of Transcription,
Dependent Nuclear

Dna-Dependent ///
Receptor

Regulation Of
Transcription

Hormone
Coactivator Activity ///

Biosynthesis
Metal Ion Binding ///

Nucleic Acid Binding

/// Dna Binding

203148_s_at
TRIM14
Tripartite Motif-
Compartment
Protein Binding ///
Cytoplasm ///

Containing 14
Specification
Zinc Ion Binding ///
Intracellular

Metal Ion Binding

210705_s_at
TRIM5
Tripartite Motif-
Protein
Ubiquitin-Protein
Ubiquitin

Containing 5
Ubiquitination ///
Ligase Activity /// Zinc
Ligase

Ubiquitin Cycle
Ion Binding /// Ligase
Complex ///

Activity /// Metal Ion
Intracellular

Binding

220558_x_at
TSPAN32
Tetraspanin 32
Cell-Cell Signaling
—
Integral To

Membrane ///

Integral To

Membrane

1557073_s_at
TTBK2
Tau Tubulin
Protein Amino Acid
Nucleotide
Intermediate

Kinase 2
Phosphorylation
Binding ///
Filament

Protein Kinase

Activity /// Atp

Binding ///

Kinase Activity

/// Transferase

Activity ///

Structural

Molecule

Activity

202335_s_at
UBE2B
Ubiquitin-
Dna Repair ///
Ubiquitin-Protein
Nucleus ///

Conjugating
Ubiquitin Cycle ///
Ligase Activity
Membrane

Enzyme E2B
Protein Modification
/// Ubiquitin-Like

(Rad6 Homolog)
/// ResponseTo Dna
Activating

Damage Stimulus
Enzyme Activity

/// Ligase

Activity

200668_s_at
UBE2D3
Ubiquitin-
Ubiquitin Cycle ///
Ubiquitin-Protein
—

Conjugating
ProteinModification
Ligase Activity

Enzyme E2D 3

/// Protein

(Ubc4/5

Binding ///

Homolog,

Ubiquitin-Like

Yeast)

Activating

Enzyme Activity

/// Ligase

Activity

215737_x_at
USF2
Upstream
Regulation Of
Transcription
Nucleus

Transcription
Transcription, Dna-
Factor Activity ///

Factor 2, C-Fos
Dependent ///
Rna Polymerase

Interacting
Transcription ///
Ii Transcription

Regulation Of
Factor Activity ///

Transcription
Dna Binding ///

Transcription

Regulator

Activity

201557_at
VAMP2
Vesicle-
Vesicle-Mediated
—
Integral To

Associated
Transport

Membrane ///

Membrane

Synaptosome

Protein 2

/// Synapse

(Synaptobrevin

2)

204254_s_at
VDR
Vitamin D (1,25-
Transcription
Transcription
Nucleus

Dihydroxyvitamin D3)
/// Regulation
Factor Activity ///

Receptor
Of
Steroid Hormone

Transcription,
Receptor Activity ///

Dna-
Protein Binding ///

Dependent ///
Vitamin D3

Signal
Receptor Activity ///

Transduction
Metal Ion Binding ///

/// Negative
Dna Binding ///

Regulation
Protein Binding ///

Of
Dna Binding ///

Transcription
Receptor Activity ///

Ligand-Dependent

Nuclear Receptor

Activity /// Zinc Ion

Binding /// Dna

Binding

217234_s_at
VIL2
Villin 2 (Ezrin)
Cytoskeletal
Structural Molecule
Cytoplasm

Anchoring ///
Activity ///
///

Regulation
Cytoskeletal
Cytoskeleton

Of Cell
Protein Binding ///
/// Microvillus

Shape
Protein Binding ///
///

Binding
Membrane

/// Actin

Filament ///

Cortical

Cytoskeleton

1562955_at
WDFY1
Wd Repeat And Fyve
—
Phosphatidylinositol
Nucleus ///

Domain Containing 1

Binding /// Zinc Ion
Early

Binding /// Metal Ion
Endosome

Binding /// Zinc Ion
/// Cytosol

Binding

208743_s_at
YWHAB
Tyrosine 3-
—
Monooxygenase
—

Monooxygenase/Tryptophan

Activity /// Protein

5-Monooxygenase

Domain Specific

Activation Protein, Beta

Binding /// Protein

Polypeptide

Binding /// Protein

Binding

217741_s_at
ZA20D2
Zinc Finger, A20 Domain
—
Dna Binding /// Zinc
—

Containing 2

Ion Binding /// Metal

Ion Binding

222357_at
ZBTB20
Zinc Finger And Btb Domain
Transcription
Dna Binding ///
Nucleus

Containing 20
/// Regulation
Protein Binding ///

Of
Zinc Ion Binding ///

Transcription,
Metal Ion Binding

Dna-

Dependent

219062_s_at
ZCCHC2
Zinc Finger, Cchc Domain
—
Nucleic Acid
—

Containing 2

Binding /// Metal Ion

Binding /// Zinc Ion

Binding

A number of genes associated with viral response, cellular defense, and immune response genes were identified. A representative list of genes in the signature set is given in Table 3.

TABLE 3

Representative genes in the signature set of chronic HCV infection:

Gene

GO Biological

Probe Set ID
Symbol
Gene Title
Process

201642_at
IFNGR2
Interferon Gamma Receptor 2
Response to virus

202086_at
MX1
Myxovirus (Influenza Virus) Resistance 1
Response to virus

202430_s_at
PLSCR1
Phospholipid Scramblase 1
Response to virus

203882_at
ISGF3G
Interferon-Stimulated Transcription Factor 3,
Response to virus

Gamma 48 Kda

204994_at
MX2
Myxovirus (Influenza Virus) Resistance 2
Response to virus

208436_s_at
IRF7
Interferon Regulatory Factor 7
Viral induction of host

immune response,

Response to virus

1553530_a_at
ITGB1
Integrin, Beta 1 (Fibronectin Receptor, Beta
Cellular defense

Polypeptide, Antigen Cd29 Includes Mdf2, Msk12)
response

1553530_a_at
ITGB1
Integrin, Beta 1 (Fibronectin Receptor, Beta
Cellular defense

Polypeptide, Antigen Cd29 Includes Mdf2, Msk12)
response

1555832_s_at
KLF6
Kruppel-Like Factor 6
B cell differentiation,

regulation of

transcription, DNA-

dependent

200959_at
FUS
Fusion (Involved In T(12; 16) In Malignant
Immune response

Liposarcoma)

201762_s_at
PSME2
Proteasome (Prosome, Macropain) Activator
Immune response

Subunit 2 (Pa28 Beta)

201786_s_at
ADAR
Adenosine Deaminase, Rna-Specific
Antimicrobial humoral

response (sensu

Vertebrata)

202086_at
MX1
Myxovirus (Influenza Virus) Resistance 1,
Immune response,

Interferon-lnducible Protein P78 (Mouse) ///
response to virus

Myxovirus (Influenza Virus) Resistance 1,

Interferon-Inducible Protein P78 (Mouse)

GO = Gene Ontology

Example 5
VX-950 Normalizes the Signature Set Over the 14-Day Treatment Period

There was an observable trend in the gene expression levels normalizing towards healthy subject levels on dosing with VX-950. Delta expression levels were calculated as the mean ratio of interferon (IFN)-sensitive gene (ISG) expression levels for each patient (day 14 vs. day 0) shown on a log₁₀scale. Delta viral load was calculated as the ratio of viral load for each patient (day 0 vs. day 14) shown on a log₁₀scale. The correlation with healthy subject levels was determined for healthy subjects after 5 days of dosing with VX-950, and for HCV infected patients at pre-dosing, and after 7, 14, and 28 days of dosing with VX-950. The results are shown in FIG. 2.

Example 6
HCV Infection Enriches for Genes of Host Anti-Viral Gene Categories

In HCV infected subjects, the gene expression analysis revealed a significant over-representation of gene ontology (GO) categories related to host response to viral infection (Table 4). Also observed was a significant enrichment for known interferon-sensitive genes (ISG) (p<10⁻⁶) (where the p-value represents the probability that the enrichment of the genes in that functional category is random.)

TABLE 4

Signature set enriched for host anti-viral GO categories:

# Genes
# Genes on

Gene Ontology category
p-value
altered
genechip

Immune response
3.4 × 10⁻⁷
30
566

Response to biotic stimulus
4.1 × 10⁻⁸
36
705

Response to stimulus
7.7 × 10⁻⁸
50
1230

Defense response
7.5 × 10⁻⁷
31
620

Response to pest, pathogen
1.3 × 10⁻⁴
19
378

or parasite

Response to stress
1.0 × 10⁻⁵
31
701

Response to virus
4.5 × 10⁻⁴
6
51

Other genes in the signature set mapped to host immune response functions and other key biological functions related to a host of anti-viral defense mechanisms. For example, the genes mapped to functions related to organismal physiological processes; immune response; defense response; response to biotic stimulus; response to external stimulus; response to stimulus; response to external biotic stimulus; response to stress; response to pest, pathogen, or parasite; response to virus.

Example 7
Pre-Dose Expression Levels of IFN-Sensitive Genes Correlates with a Reduction in Plasma HCV RNA Levels

Table 5 shows the ratios of IFN-sensitive gene (ISG) expression levels between the enhanced responders and non-enhanced responders (the ratio is the level of expression of the enhanced responders over the levels of expression of the non-enhanced responders) prior to dosing with VX-950. The pre-dose expression levels of these genes correlates with plasma HCV RNA reduction.

TABLE 5

Ratios of ISG levels Between Enhanced Responders and Others

Affymetrix

Gene
GO Biological

Probeset ID
Gene Title
Symbol
Process Description
Ratio

203153_at
Interferon-Induced Protein With Tetratricopeptide Repeats 1
IFIT1
Immune Response
8.57

204439_at
Interferon-Induced Protein 44-Like
IFI44L
—
4.17

213797_at
Radical S-Adenosyl Methionine Domain Containing 2
RSAD2
—
4.11

226757_at
Interferon-Induced Protein With Tetratricopeptide Repeats 2
IFIT2
Immune Response
3.48

204747_at
Interferon-Induced Protein With Tetratricopeptide Repeats 3
IFIT3
Immune Response
2.91

206332_s_at
Interferon, Gamma-Inducible Protein 16
IFI16
Immune Response,
2.79

DNA-dependent Regulation Of

Transcription

208966_x_at
Interferon, Gamma-Inducible Protein 16
IFI16
Immune Response,
2.75

DNA-dependent Regulation Of

Transcription

214453_s_at
Interferon-Induced Protein 44
IFI44
Immune Response
2.73

217502_at
Interferon-Induced Protein With Tetratricopeptide Repeats 2
IFIT2
Immune Response
2.73

203595_s_at
Interferon-Induced Protein With Tetratricopeptide Repeats 5
IFIT5
Immune Response
2.68

229450_at
Interferon-Induced Protein With Tetratricopeptide Repeats 3
IFIT3
Immune Response
2.46

208965_s_at
Interferon, Gamma-Inducible Protein 16
IFI16
Immune Response,
2.45

DNA-dependent Regulation Of

Transcription

203596_s_at
Interferon-Induced Protein With Tetratricopeptide Repeats 5
IFIT5
Immune Response
1.69

202446_s_at
Phospholipid Scramblase 1
PLSCR1
Response To Virus,
1.42

Phospholipid Scrambling

202086_at
Myxovirus (Influenza Virus) Resistance 1
MX1
Immune Response, Signal
1.39

Transduction

202411_at
Interferon, Alpha-Inducible Protein 27
IFI27
Immune Response
1.16

209417_s_at
Interferon-Induced Protein 35
IFI35
Immune Response
1.11

201601_x_at
Interferon Induced Transmembrane Protein 1 (9-27)
IFITM1
Immune Response, Negative
1.01

Regulation Of Cell Proliferation

212203_x_at
Interferon Induced Transmembrane Protein 3 (1-8U)
IFITM3
Immune Response
1.01

201422_at
Interferon, Gamma-Inducible Protein 30
IFI30
Immune Response
0.93

214022_s_at
Interferon Induced Transmembrane Protein 1 (9-27)
IFITM1
Immune Response, Negative
0.93

Regulation Of Cell Proliferation

201315_x_at
Interferon Induced Transmembrane Protein 2 (1-8D)
IFITM2
Immune Response
0.82

Example 8
Sustained Levels of Interferon-Sensitive Genes Correlate with a Reduction in Plasma HCV RNA Levels

The expression levels of selected interferon-sensitive genes (ISGs) were examined pre-dosing and at day 14 after dosing with VX-950 in HCV-infected enhanced responders and non-enhanced responders. The mean ratios of ISG expression levels (day 14 (d14) vs. pre-dose (d0)) are shown in FIG. 3A. There was a statistically significant difference in the sustained expression levels of the ISG between the two groups, wherein the enhanced responders had sustained levels of ISG expression. Genes that were outliers within each group are listed. Thus, in as little as 14 days, a comparison of baseline to day 14 expression levels of ISGs can potentially predict VX-950 dosing outcomes.

FIG. 3B shows the change in expression levels and change in HCV viral load by day 14 as compared to day 0 in five enhanced responders (left-most bars) and 16 non-enhanced responders. The five enhanced responders, who had undetectable HCV RNA at day 14, had sustained levels of the IFN-sensitive genes (ISGs), as indicated by the minimal change in their expression levels.

FIG. 3C shows quantitative real-time PCR confirmation of the Affymetrix genechip results. Gene expression modulation of specific ISGs for each of the groups in 3B are shown (top left panel shows the results for the enhanced responders while the top right and bottom panels show the results for the non-enhanced responders). The overall trend confirms the genechip profiling data. There are also individual gene-level expression differences (e.g., GIP2, PLSCR) between the enhanced and non-enhanced responders.

From these results, it appears that sustained levels of interferon-induced genes in peripheral blood during VX-950 dosing were associated with best antiviral response.

Example 9
Signature Sets of Specific HCV Subgroups

The signature set shown in Table 2 was obtained from a population of chronically infected HCV subjects without a priori bias using a unsupervised clustering method. A signature set for a selected group can be prepared based on the teachings provided herein. For example, a signature set can be generated for certain subgroups of HCV-infected subjects, for example: males, females, HCV genotype 1, 2, or 3, particular age groups, races, subjects that have responded well or poorly to previous treatments, subjects who have previously undergone a particular treatment, subjects who have not yet undergone treatment for HCV infection, subjects who have been diagnosed as being co-infected with another virus (e.g., hepatitis B and/or HIV), etc.

The information obtained from such analyses can be utilized as described herein.

A number of embodiments of the invention have been described. Nevertheless, it will be understood that various modifications may be made without departing from the spirit and scope of the invention. Accordingly, other embodiments are within the scope of the following claims.

HEPATITIS C VIRUS INFECTION BIOMARKERS

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Date Filed

Date Published

Inventors

CPC

US Classifications

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Abstract

Description

Claims

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