JP2011155866A - Method for predicting therapeutic effect of chronic type c hepatitis - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a method for predicting the effect of combined therapy by interferon and ribavirin without requiring complicated operation, especially the method for predicting the effect of the combined therapy by the interferon and the ribavirin without actually administering them. <P>SOLUTION: The method for predicting the effect of the combined therapy of the interferon and the ribavirin against chronic type C hepatitis includes a step for measuring the expression level of a biological material in a cell of a carrier of a type C hepatitis virus before administering the interferon and the ribavirin to the carrier of the type C hepatitis virus, wherein the biological material is at least one selected from the group consisting of MX1, IFIT1, ISG15 and IFI27. <P>COPYRIGHT: (C)2011,JPO&INPIT

Description

  The present invention relates to a method for predicting the effect of combined treatment with interferon and ribavirin for chronic hepatitis C caused by hepatitis C virus, a method for predicting recurrence after combined treatment, and a kit therefor.

Chronic hepatitis C (hereinafter sometimes referred to as CH) is a disease caused by infection with the hepatitis C virus (HCV) belonging to the genus Pacivirus in the Flaviviridae family, and currently more than 3% of the world population Is infected with this virus.
The hepatitis C virus has a spherical single-stranded RNA of about 9.5 kb in the form of a sphere having a diameter of 55 to 57 nm as a genome, a viral gene and a nucleocapsid (core particle) enclosing it, and It consists of a double structure of an envelope.
Chronic hepatitis C is one of the diseases formerly called non-A non-B hepatitis. In 1989, Choo et al. Inoculated patients' plasma into chimpanzees and cloned viral genes from the plasma. Hepatitis C virus was identified for the first time (Non-patent Document 1).

  Hepatitis C virus has a plurality of genotypes and is currently classified as type 1a, 1b, 2a, 2b, 3a, 3b, 4, 5a, 6a. Among these genotypes, in Japan, type 1b, which is considered difficult to treat with interferon, accounts for 70-85%. Thereafter, 2a type is 10-15%, 2b type is about 5%, and other genotypes are rare.

  Hepatitis C virus rarely becomes fulminant due to primary infection, but after acute hepatitis onsets, the case where the alanine aminotransferase (ALT) level is maintained at a high level and the virus also persists to transition to chronic hepatitis And may be an asymptomatic carrier. It is reported that about 70% of the first infected persons become carriers and become chronic. Also, chronic hepatitis C rarely heals spontaneously, and 30-50% of patients develop cirrhosis between 20-30 years after infection. In addition, 60-80% of patients with cirrhosis develop liver cancer.

  Interferon has been used as a therapeutic method for chronic hepatitis C, but the efficiency was only about 30%. However, in recent years, combined treatment with interferon and ribavirin, which is one of antiviral drugs, has been performed.

At present, combined treatment of polyethyleneglycolized interferon and ribavirin, which has a long half-life in blood by binding polyethylene glycol to interferon, is the standard treatment method. However, in the hepatitis C virus infected by Japanese, in the case of high viral load of type 1b infection, which is the most common genotype, the ratio of the most effective cases is about 50-60% even with the above combination treatment It is not effective for about half of patients.
Furthermore, for example, the drug administration period of the combined treatment of polyethylene glycolated interferon alpha-2b and ribavirin ranges from 48 to 72 weeks. During this period, fever, headache, malaise, loss of appetite, itching of the body, hair loss, thrombocytopenia The frequency of such side effects is also high. In addition, because of the high drug price, combined treatment with interferon and ribavirin cannot ignore the physical, mental and economic burdens of patients.

Therefore, if it can be determined before treatment whether combined therapy of interferon and ribavirin is effective, treatment with side effects can be avoided for patients who are not effective for combined therapy. It is possible to administer other treatment methods.
Hepatitis C virus core contained in a sample collected from a patient infected with genotype 1b hepatitis C virus as a method for determining the effectiveness of combined treatment of interferon and ribavirin for the treatment of chronic hepatitis C It has been proposed to check the amino acid sequence in the region, whether the 70th arginine is mutated to another amino acid and whether the 91st amino acid is methionine (Patent Document 1). However, in this method, it is necessary to carry out the identification of mutated amino acids in the virus, which is very complicated. In addition, this method cannot predict recurrence after treatment.

In addition, in patients infected with hepatitis C virus, ANXA3, PCNA, LMNB1, PAK1, 4EBP1 before and after the combined treatment with interferon and ribavirin in order to examine the effect of the combined treatment with interferon and ribavirin. , KARS, PSME2, ENO1, STAT1, PHB, SP-100, and PPP1CB are measured for the expression level of specific biological substances, and that the difference in expression level is found that the effectiveness of combined treatment with interferon and ribavirin A technique for indicating the index has been developed (Patent Document 2). However, in this method, in order to predict the therapeutic effect, it is necessary to measure the expression level twice before and after administration of interferon and ribavirin, and the work is complicated. In addition, when interferon and ribavirin are administered directly to a subject, the risk of serious side effects cannot be excluded.
Further, the above-described methods are methods for predicting effective cases of combined treatment of interferon and ribavirin, and are not methods for actively predicting ineffective cases in order to avoid the economic burden and side effects of treatment.

JP 2007-43985 A JP 2009-178057 A

Choo QL, et al. , Science. 244, 359-362 (1989)

  As described above, there has been no report yet on a method for predicting the effect of combined treatment with interferon and ribavirin without requiring complicated work.

  Therefore, the present invention predicts the effect of combined treatment of interferon and ribavirin without requiring complicated work, and in particular, predicts the effect of combined treatment without actually administering interferon and ribavirin. It aims to provide a method. Another object of the present invention is to provide a method for predicting the presence or absence of recurrence after combined treatment with interferon and ribavirin.

As a result of intensive investigations to solve the above problems, the present inventors discriminate treatment effective cases, relapse cases, and treatment invalid cases with high probability by measuring the expression levels of some innate immunity related genes. Developed a method.
That is, the present invention includes a step of measuring an expression level of a biological substance in a cell of the hepatitis C virus carrier before administering interferon and ribavirin to the hepatitis C virus carrier, Is a method for predicting the effect of a combined treatment of interferon and ribavirin on chronic hepatitis C, which is at least one selected from the group consisting of MX1, IFIT1, ISG15, and IFI27.

In addition, the present invention includes a step of measuring an expression level of a biological material in a cell of the hepatitis C virus carrier before administering interferon and ribavirin to the hepatitis C virus carrier, Is a combination therapy of interferon and ribavirin for chronic hepatitis C, wherein is at least one selected from the group consisting of PSMB10, TRAF2, RXRA, OASI, IKK / NEMO, MyD88, IFITM3, IL10RB, RNF125, BCL3, NFKB2 It is a method for predicting later recurrence.
The present invention also includes SEQ ID NO: 14, SEQ ID NO: 21, SEQ ID NO: 37, SEQ ID NO: 59, SEQ ID NO: 74, SEQ ID NO: 100, SEQ ID NO: 101, SEQ ID NO: 107, SEQ ID NO: 115, SEQ ID NO: 120, SEQ ID NO: 135, Interferon and ribavirin for chronic hepatitis C, comprising at least one of the nucleotide sequence shown in SEQ ID NO: 150, SEQ ID NO: 184, SEQ ID NO: 193 or SEQ ID NO: 234, or an oligonucleotide comprising a nucleotide sequence complementary thereto It is a kit for predicting the effect of the combination therapy or recurrence after the combination therapy of interferon and ribavirin.

According to the prediction method of the present invention, it is possible to efficiently determine the effect of the combination treatment, which has not been known until now by actually administering interferon and ribavirin, using only gene expression information before administration. In addition, the presence or absence of recurrence after combination treatment can be efficiently determined using only gene expression information before administration. For this reason, the effect of combined treatment and the recurrence after combined treatment can be predicted without requiring complicated work.
With the prediction method of the present invention, hepatitis treatment with a good prospect of avoiding side effects can be performed. Since treatment invalid cases and relapse cases can be predicted with high probability, it is possible to select a treatment method according to the genetic information of the individual patient. For example, patients who are expected to be ineffective can avoid the side effects of administration and can be judged to extend the treatment period in cases where recurrence is expected. .
Furthermore, since the presence or absence of treatment effects can be predicted before the start of combination treatment, it is possible to avoid costly burdens on patients and society by administering expensive drugs such as interferon. Useful for cost reduction.

FIG. 1 is a diagram showing an outline of the whole experiment. FIG. 2 shows the correspondence between the prediction of the combined treatment effect of interferon and ribavirin and the actual diagnosis result. The lower part of FIG. 2 is a diagram showing the correspondence between the prediction of the presence or absence of recurrence and the actual case of recurrence among cases having a combined treatment effect.

Hepatitis C virus carrier In the present invention, when referring to a hepatitis C virus carrier , a genotype of any virus of type 1a, 1b, 2a, 2b, 3a, 3b, 4, 5a, or 6a The subject infected with is not specifically limited.

Interferon and ribavirin combination therapy Interferon and ribavirin combination therapy is a treatment method for chronic hepatitis C in which ribavirin is administered together with interferon. The type of interferon used for the combination therapy is not particularly limited, and any interferon used for the treatment of chronic hepatitis C may be used. Usually, interferon α, particularly interferon α2a and interferon α2b are used, but not limited thereto. In order to increase the blood half-life of interferon, it is desirable to use interferon (polyethylene glycolated interferon) combined with polyethylene glycol.

  Regarding the above combination therapy, according to the guidelines for the treatment of chronic hepatitis C in 2009, in the case of genotype 1b hepatitis C virus, the serum hepatitis C virus RNA concentration is 5.0 Log IU / mL, or 300 fmol / In the case of a high viral load of L or more, a combined treatment of polyethylene glycolated interferon α2a and ribavirin, or polyethylene glycolated interferon α2b and ribavirin is supposed to be performed for 48 to 72 weeks. When the RNA concentration of hepatitis C virus in serum is 5.0 Log IU / mL or a low viral load of less than 300 fmol / L, polyethylene glycolated interferon α2a is to be administered for 24 to 48 weeks. In addition, the combination treatment which can predict the effect and the presence or absence of recurrence by the method of the present invention is not necessarily limited to the combination treatment according to these guidelines, and may be a combination treatment of other interferon and ribavirin.

The effect of treatment The actual treatment effect can be judged by whether or not the amount of RNA of hepatitis C virus in the blood is below the detection sensitivity after the combined treatment of interferon and ribavirin. Hepatitis C virus can be detected and quantified by performing PCR, real-time PCR, bDNA method, etc. on RNA derived from the virus. Real-time PCR is the best in both detection sensitivity and measurement range, but the viral load evaluation method as a criterion for judging the therapeutic effect is not particularly limited.
In addition, "therapeutic effect" said here also includes a case of recurrence in the future, and transiently, after combined treatment with interferon and ribavirin, RNA of hepatitis C virus in the blood The amount is below the detection sensitivity.

  Therefore, the method for predicting the effect of the combination treatment in the present invention is a person who temporarily loses the amount of hepatitis C virus RNA during detection of the combination treatment, or after the combination treatment, or temporarily, This method predicts whether hepatitis C virus RNA will continue to be detected throughout.

Recurrence and recurrence means that the combined treatment of interferon and ribavirin reduces the amount of viral RNA below the detection sensitivity, but after the combined treatment is completed, viral RNA is detected again.

  Therefore, the method for predicting recurrence in the present invention is a person whose viral RNA amount is below the detection sensitivity due to the combination therapy, or a person whose viral RNA is detected again after the combination therapy is completed or is not detected again. It is a method of predicting whether it is.

Any tissue may be used as long as it is collected from a carrier of cellular hepatitis C virus. For example, peripheral blood-derived cells and liver-derived cells (liver cells) are preferable, but liver-derived cells with good sensitivity are particularly preferable.

Biological material A biological material is a chemical substance that exists in the body of a living organism, and here refers to a biopolymer that is a basic material that constitutes a living body, or a biopolymer that is artificially synthesized based on these. When measuring multiple biological materials, it is preferable to measure the amount of mRNA or nucleic acid synthesized and amplified based on it as the biological material because of the economics and simplicity of the detection method. Can also measure proteins.

When measuring the expression level of a specific mRNA, such as PCR, Real-Time PCR, Northern blotting, DNA microarray, etc. When measuring the expression level of a specific protein, Western blotting or ELISA Various methods known to those skilled in the art can be used as long as the method is specific to a biological substance to be detected, such as an antibody array.
The measurement of the expression level of these biological substances may be performed by measuring the expression level of the biological material modified by reverse transcription or amplification by enzymatic treatment.
The expression level of the biological material can be compared by detecting and digitizing a signal correlated with the amount of the biological material. As the signal, a fluorescent substance, a radioactive substance, an enzyme, or any other signal known to those skilled in the art can be used.

MX1
MX1 may be anything including a part of the mRNA sequence shown in GeneBank accession number NM_002462 and its complementary sequence, and a part of the amino acid sequence encoded by it. Is preferably a sequence comprising all or part of the nucleic acid sequence set forth in SEQ ID NO: 100 or its complementary sequence.

IFIT1
IFIT1 may be anything including part of the mRNA sequence shown in GeneBank accession number NM_001548, its complementary sequence, and part of the amino acid sequence encoded by it. Is preferably a sequence comprising all or part of the nucleic acid sequence set forth in SEQ ID NO: 135 or its complementary sequence.

ISG15
ISG15 may be anything including part of the mRNA sequence shown in GeneBank accession number NM_005101, its complementary sequence, and part of the amino acid sequence encoded by it. Is preferably a sequence containing all or part of the nucleic acid sequence set forth in SEQ ID NO: 107 or its complementary sequence.

IFI27
IFI27 may be anything including a part of the mRNA sequence shown in GeneBank accession number NM_005532 and its complementary sequence, and a part of the amino acid sequence encoded by it. Is preferably a sequence containing all or part of the nucleic acid sequence set forth in SEQ ID NO: 120 or its complementary sequence.

PSMB10
PSMB10 may be anything including part of the mRNA sequence indicated by GeneBank accession number NM_002801, its complementary sequence, and part of the amino acid sequence encoded by it. Is preferably a sequence containing all or part of the nucleic acid sequence set forth in SEQ ID NO: 150 or its complementary sequence.

TRAF2
TRAF2 may be anything including a part of mRNA sequence shown in GeneBank accession number NM_021138, its complementary sequence, and part of the amino acid sequence encoded by it. Is preferably a sequence containing all or part of the nucleic acid sequence set forth in SEQ ID NO: 21 or its complementary sequence.

RXRA
RXRA may be anything including a part of the mRNA sequence shown in GeneBank accession number NM_002957, its complementary sequence, and a part of the amino acid sequence encoded by it. Is preferably a sequence comprising all or part of the nucleic acid sequence set forth in SEQ ID NO: 234 or its complementary sequence.

OASI
OASI may be anything including part of the mRNA sequence shown in GeneBank Accession No. NM — 016816, its complementary sequence, and part of the amino acid sequence encoded by it. Is preferably a sequence comprising all or part of the nucleic acid sequence set forth in SEQ ID NO: 101 or its complementary sequence.

IKKG / NEMO
IKKG / NEMO may be anything including a part of the mRNA sequence shown in GeneBank accession number NM_001099857, its complementary sequence, and a part of the amino acid sequence encoded by it. Specifically, it is desirable that the sequence includes all or part of the nucleic acid sequence shown in SEQ ID NO: 59 or its complementary sequence.

MyD88
MyD88 may be anything including a part of the mRNA sequence shown in GeneBank accession number NM_002468, its complementary sequence, and a part of the amino acid sequence encoded by it. Is preferably a sequence containing all or part of the nucleic acid sequence shown in SEQ ID NO: 14 or its complementary sequence.

IFITM3
IFITM3 may be anything including a part of the mRNA sequence shown in GeneBank accession number NM — 021034, its complementary sequence, and a part of the amino acid sequence encoded by it. Is preferably a sequence containing all or part of the nucleic acid sequence set forth in SEQ ID NO: 115 or its complementary sequence.

IL10RB
IL10RB may be anything including part of the mRNA sequence shown in GeneBank accession number NM_000628 and its complementary sequence, and part of the amino acid sequence encoded by it. Is preferably a sequence containing all or part of the nucleic acid sequence set forth in SEQ ID NO: 37 or its complementary sequence.

RNF125
RNF125 may be anything including part of the mRNA sequence shown in GeneBank accession number NM_017831, its complementary sequence, and part of the amino acid sequence encoded by it. Is preferably a sequence containing all or part of the nucleic acid sequence set forth in SEQ ID NO: 193 or its complementary sequence.

BCL3
BCL3 may be anything including a part of the mRNA sequence shown in GeneBank accession number NM_005178, its complementary sequence, and a part of the amino acid sequence encoded by it. Is preferably a sequence comprising all or part of the nucleic acid sequence set forth in SEQ ID NO: 74 or its complementary sequence.

NFKB2
NFKB2 may be anything including part of the mRNA sequence shown in GeneBank accession number NM_002502, its complementary sequence, and part of the amino acid sequence encoded by it. Is preferably a sequence containing all or part of the nucleic acid sequence set forth in SEQ ID NO: 184 or its complementary sequence.

Predictive method After measuring the expression level of the biological material, the expression level of each biological material is compared with the representative value of each group accumulated in advance, and by examining which of them is close, The effect of combination treatment with interferon and ribavirin can be predicted, and the possibility of recurrence after treatment can be evaluated. In this case, any statistical method may be used as long as the expression level can be compared for each group. In order to investigate which of the representative values of each group the expression level of the biological material is included, any clustering method known to those skilled in the art can be used.

Kit In order to carry out the above prediction easily, it is also possible to create the following kit with a simpler detection method.
That is, as a kit for predicting the effect of combined treatment with interferon and ribavirin in a hepatitis C virus carrier, at least 10 consecutive nucleotides contained in the base sequences derived from MX1, IFIT1, ISG15, and IFI27, Or a kit for detecting the effect of a combined treatment of interferon and ribavirin, comprising at least one, preferably two or more oligonucleotides comprising at least 10 nucleotides of a base sequence complementary thereto. .
These are typically oligonucleotides comprising at least 10 consecutive nucleotides contained in the nucleotide sequences shown in SEQ ID Nos. 100, 135, 107, and 120 of the sequence listing, or at least 10 nucleotides complementary to them. A kit for detecting the effect of a combined treatment of interferon and ribavirin in a hepatitis C virus carrier comprising at least one.

In addition, as a kit for predicting recurrence after combined treatment with interferon and ribavirin in hepatitis C virus carriers, PSMB10, TRAF2, RXRA, OASI, IKKG / NEMO, MyD88, IFITM3, IL10RB, RNF125, BCL3 An interferon comprising at least one, preferably two or more of oligonucleotides comprising at least 10 consecutive nucleotides contained in the nucleotide sequence derived from NFKB2 or at least 10 nucleotides complementary to the nucleotide sequence, respectively, A kit is provided for predicting recurrence after combination therapy with ribavirin.
These are typically at least 10 consecutive nucleotides contained in the base sequence shown in SEQ ID NO: 150, 21, 234, 101, 59, 14, 115, 37, 193, 74, 184 of the sequence listing, or A kit for predicting recurrence after combined treatment of interferon and ribavirin in a hepatitis C virus carrier, comprising at least one oligonucleotide comprising at least 10 nucleotides of complementary base sequence.
By using the oligonucleotide contained in such a kit as a probe, for example, Northern blotting technique is used to analyze mRNA in the cell, thereby making it possible to easily examine the amount of target RNA contained in the cell.
Furthermore, efficient detection can be performed by preparing a DNA microarray in which a plurality of these oligonucleotides are immobilized on a solid support. In addition to the nucleotides described above, it is desirable that such a DNA microarray is loaded with a plurality of types of genes related to innate immunity as shown in Tables 1 to 7 below and other control genes. .

Preparation of sample Hepatitis C virus among patients confirmed to be not applicable to autoimmune hepatitis, alcoholic liver injury, hepatitis B virus related antigen / antibody or anti-HIV antibody detected Targeted 87 patients with chronic hepatitis C who had been infected with genotype 1b and had not been treated with interferon or immunomodulation in the past, and 10 patients with fatty liver who were not infected with hepatitis C virus .
Prior to combined treatment with interferon and ribavirin, hepatitis C virus RNA in serum was quantified using Amplicor-HCV Monitor Assay (Roche). In addition, aspartate aminotransferase (AST), alanine aminotransferase (ALT), total bilirubin, alkaline phosphatase (ALP), gamma glutamyl transpeptidase (γ-GTP), white blood cells, platelets, hemoglobin, and albumin were measured.
The histological staging of liver biopsy specimens collected from patients with chronic hepatitis C was performed according to the Metaville classification system.

Protocol for combined treatment of interferon and ribavirin All patients with chronic hepatitis C were treated with a combination treatment of polyethyleneglycolized interferon alfa 2b (Schering-Plough) and ribavirin (Schering-Plough) for 48 weeks. . Polyethylene glycolated interferon alpha 2b was administered at 1.5 μg per week per kilogram of body weight at the start of drug administration, and ribavirin was administered according to the recommended amount disclosed by the manufacturer.

Definition of Drug Response First, patients who did not detect hepatitis C virus RNA in serum at 4 weeks after drug administration were classified into two groups as Rapid Virtual Responder (RVR) and the others as nonRVR.
Next, patients who had no hepatitis C virus RNA detected in the serum at 12 weeks after drug administration were treated as Complete Early Virtual Responder (cEVR), and hepatitis C virus RNA in the serum at 12 weeks after drug administration. Patients who were detected but whose amount was reduced by 2 orders of magnitude or more compared to before administration of the drug were classified into 3 groups as Partial EVR (pEVR), and the other patients as nonEVR.

  Sustained virological responder (SVR) for patients whose hepatitis C virus RNA was not detected for 6 months after the end of combination therapy, and patients who were detected again after hepatitis C virus RNA was not detected at the end of combination therapy Patients with continued detection of hepatitis C virus throughout the treatment with Relax (R) were classified into 3 groups as Non-Virological Responder (NR).

RNA extraction RNA was extracted from cells derived from a liver biopsy specimen, which is a kind of liver-derived cells (liver cells). Liver biopsy specimens were placed in RNA Later (Ambion) until RNA was extracted and stored at -80 ° C. Total RNA was extracted using mirVana (TM) miRNA Isolation kit according to the instructions disclosed by the manufacturer.

Acquisition of expression level data by DNA microarray Using DNA chip Genopearl manufactured by Mitsubishi Rayon Co., Ltd., 237 types of innate immunity-related genes listed in Tables 1 to 7 (200 types for Chip1, 37 types for Chip2) were mounted. A DNA microarray was prepared.

  Total RNA obtained from the liver biopsy specimen was subjected to biotin labeling and amplification reaction using MessageAmp-Biotin Enhanced Kit (Ambion) according to the instructions disclosed by the manufacturer. The obtained biotin-labeled RNA (5 μg) was fragmented by heating at 94 ° C. for 7 minutes and 30 seconds using the Fragmentation Buffer attached to the kit.

Various reagents were added to the fragmented biotin-labeled RNA solution so that the final composition was 0.12M Tris-HCl / 0.12M NaCl / 0.05% Tween 20, and 150 μl of a hybridization sample was obtained.
The DNA microarray and the hybridization sample were reacted at 65 ° C. for 16 hours, and then washed twice with a washing solution (0.12M Tris-HCl / 0.12M NaCl / 0.05% Tween 20) at 65 ° C. for 20 minutes. . Further, the DNA microarray was subjected to liquid replacement for 10 minutes at 65 ° C. with a storage solution (0.12 M Tris-HCl / 0.12 M NaCl).

  The washed DNA microarray was immersed in Streptavidin-Cy5 (GE Healthcare BioScience KK) dissolved in 2 μg / ml with a stock solution for 30 minutes at room temperature, and then washed with the washing solution for 5 minutes at room temperature for 4 times. The solution was replaced with the preservation solution for 5 minutes.

  The DNA microarray after Cy5 labeling uses a dedicated DNA microarray detector to detect fluorescence at a plurality of exposure times from 0 seconds to 40 seconds. The fluorescence signal intensity was quantified using the image with the longest exposure time.

Statistical analysis method The numerical data analysis between the groups divided according to the definition of the drug response was statistical analysis using Student's t-test. The Kruskal-Willis test or Fisher's exact test was used for comparison of each component in serum among the three groups of SVR, R, and NR. Moreover, in any test, when P value was less than 0.05, it was considered that there was a statistically significant difference.

Results First, average values were shown for clinical characteristics of patients with SVR, R, NR and fatty liver (Table 8).

The characteristic gene expression patterns between chronic hepatitis C patients and fatty liver patients were examined as follows. In addition, for chronic hepatitis C patients, the effect of the combined treatment was confirmed at the 4th and 12th weeks after the start of the treatment and after the end of the treatment, and a characteristic gene expression was determined for each hepatitis C virus reaction state at each time point. The pattern was examined (Figure 1).
First, gene expression patterns that were characteristically different between samples of patients with chronic hepatitis C and fatty liver were examined, and a significant difference was observed in the average signal intensity of 53 types of genes (Table 9).

  Among chronic hepatitis C patients, there was a significant difference in the average signal intensity in 14 genes between the SVR and NR samples (Table 10).

  Among chronic hepatitis C patients, there was a significant difference in the signal intensity average value among the RVR and nonRVR samples in the probes corresponding to the four genes (Table 11).

  Among chronic hepatitis C patients, a significant difference was observed in the average signal intensity of the probes corresponding to 14 types of genes between the cEVR and nonEVR samples (Table 12).

The final therapeutic effect of the combination treatment was evaluated using LOOCV (Leave-One-Out Cross Validation) (see Alain Dupuy, et al., JNCI. 99, 147-157 (2007)). First, the method for distinguishing NR from all patients with chronic hepatitis C was confirmed using the effect prediction LOOCV using two-group discrimination. When evaluated using MX1, IFIT1, ISG15, and IFI27, it was accurate. The sex, sensitivity, and specificity could be discriminated at 82.8%, 73.9%, and 85.9%, respectively (Table 13 and FIG. 2). As mentioned above, it became possible to predict the effect of combined treatment with interferon and ribavirin with high accuracy by using these genes.
Further, among those that were not NR, the method for distinguishing SVR and R was confirmed in the same manner as described above. When used and evaluated, accuracy, sensitivity, and specificity could be distinguished by 68.8%, 65.4%, and 71.1%, respectively (Table 13 bottom and FIG. 2 bottom). Based on the above, it became possible to predict recurrence after combined treatment with interferon and ribavirin with high accuracy by using these genes.

  The effect of combined treatment with interferon and ribavirin for chronic hepatitis C and recurrence after combined treatment can be predicted.

  SEQ ID NOs: 1 to 237: synthetic DNA

Claims (5)

Measuring the expression level of biological material in cells of the hepatitis C virus carrier before administering interferon and ribavirin to the hepatitis C virus carrier,
The biological material is at least one selected from the group consisting of MX1, IFIT1, ISG15, and IFI27;
A method for predicting the effect of combined treatment of interferon and ribavirin for chronic hepatitis C. Measuring the expression level of biological material in cells of the hepatitis C virus carrier before administering interferon and ribavirin to the hepatitis C virus carrier,
The biological material is at least one selected from the group consisting of PSMB10, TRAF2, RXRA, OASI, IKKG / NEMO, MyD88, IFITM3, IL10RB, RNF125, BCL3, and NFKB2.
A method for predicting recurrence after combined treatment of interferon and ribavirin for chronic hepatitis C.   The prediction method according to claim 1, wherein the cell is a liver cell.   The prediction method according to claim 1, wherein the interferon is polyethylene glycolated interferon.   Sequence number 14, Sequence number 21, Sequence number 37, Sequence number 59, Sequence number 74, Sequence number 100, Sequence number 101, Sequence number 107, Sequence number 115, Sequence number 120, Sequence number 135, Sequence number 150, Sequence number 184, the effect of the combined treatment of interferon and ribavirin for chronic hepatitis C, comprising at least one of the oligonucleotides comprising the nucleotide sequence shown in SEQ ID NO: 193 or SEQ ID NO: 234 or a nucleotide sequence complementary thereto, or Kit for predicting recurrence after combined treatment with interferon and ribavirin.

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