Exploring The Predictable Markers Of IFN Therapy In Patients With Chronic Hepatitis B

?Objective? 1To find the predictive value of single-nucleotide polymorphisms(SNPs) of host genome and the concentration of 1,25(OH)2D3 in IFN treatment for CHB patients, and provided the basis for individualized diagnosis and treatment. 2 Toinvestigate the potential value in predicting PEG-IFN therapy efficacy by monitoring the HBV markers(HBs Ag, HBe Ag) and HBV DNA levels in HBe Ag-positive CHB patients during peg-interferon(PEG-IFN) therapy and confirming the statistical threshold of each indicator.?Method?1 Exploring the predictable markers of IFN therapy in patients with chronic hepatitis BThough the Hapmap database, we got a total of 34 tagger SNPs of CYP27B1, IL-28 B and OAS gene family. Import the data into Haploview4.2 software, and linkage disequilibrium was calculated for each tagger SNPs, and ultimately selected CYP27B1(rs4646536, rs10877012), IL-28B(rs8099917) and OAS(rs10774671, rs2072138, rs2072136 and rs10849829) for experiments Compared with the polymorphisms each SNPs of 90 CHB patients, who were received IFN therapy from the the Center of Liver diseases of the First Affiliated Hospital of Fujian Medical University from May 2012 to September 2015.Patients were included 79 HBe Ag positive and 11 HBe Ag negative, and divided into three groups, complete responses(CR), partial responses(PR) and non-responses(NR). Whole blood and Serum samples were collected at baseline and week 12. The concentration of 25(OH)VD3-1?OHase and 1,25(OH)2VD3 was detected by ELISA kits. The concentration of HBV markers was tested by Chemiluminescence Microparticle Immuno Assay(CMIA), HBV DNA level was determined by real-time PCR, and ALT,AST level was measured by velocity method. The statistical analysis were performed using normality test, t test and ?2test, etc.2 Prediction of interferon treatment response by dynamically monitoring the concentration of HBV markers in HBe Ag-positive CHB patientsThis was a retrospective study,a total of 61 HBe Ag- positive CHB patients who aged 16 or older and received PEG-IFN for 48 weeks were enrolled in between October 2008 and September 2015. All of them were recruited in the Center of Liver diseases of the First Affiliated Hospital of Fujian Medical University. Based on the response to PEG-IFN therapy at the end of the study, they were divided into two groupsaccording to the treatment response at the end of the treatment, 20 complete responses(CR) and 41 partial responses(PR).CR group: HBV DNA<500 IU/m L, HBe Ag clearance with or without HBe Ag seroconversion, and HBs Ag clearance or not; PR group: the HBV DNA level decreased more than 1log10 IU/m L but still more than 500 IU/m L with HBe Ag clearance or not.Laboratory assessments were performed every 12 weeks(week12, week24, week36, and week48, respectively) from baseline to the end of the study.The concentration of HBV markers was tested by Chemiluminescence Microparticle Immuno Assay(CMIA), HBV DNA level was determined by real-time PCR, and ALT level were measured by velocity method. The statistical analysis was performed using normality test, t test and ?2test, ROC analysis, etc.?Results?1 Exploring the predictable markers of IFN therapy in patients with chronic hepatitis B1.1 The relationship between host SNPs and the efficacy of IFN treatment.At the end of treatment, for HBe Ag positive patients, 16 achieved CR, 42 achieved PR and 29 were NR; for HBe Ag negative patients, including 9 patients with CR and 2 patients with PR. Through the experiments, we got the following results: 1. CYP27B1 rs4646536(minor allele C vs major allele T, P<0.05) and rs10877012(minor allele T vs major allele G, P<0.05) were associated with non-response and lower HBe Ag clearance rates in terms of allelic frequency; so as the genotypic frequency of rs4646536(CC vs. TT/CT, P<0.05) and rs10877012(TT vs. GG/GT, P<0.05). OAS2rs2072138(major allele C vs. minor allele G, P<0.05)was independently predicted PR and the continuous positive of HBe Ag. Haplotype analysis of CYP27B1 polymorphisms showed haplotype block CT(rs4646536/rs10877012) to be associated with lower CR rates, and haplotype block TG to be associated with complete response. Haplotype analysis of OAS polymorphisms showed haplotype block AGGG(rs10774671/rs2072138/rs2072136/rs10849829) was associated with complete response and HBe Ag clearance. None of the 4 polymorphisms(rs8099917, rs10774671, rs2072136 and rs10849829) influenced the IFN efficacy. We also found that a high level of 1,25(OH)2VD3(?39.39 pg/ml) independently predict complete response.1.2The preliminary Study inthe concentration of HBs Ag, HBe Ag and HBV DNA at baseline and week 12, which were associated with IFN efficacy in CHB patients.The lower concentration of HBeAg and HBV DNA was showed in CR group than PR group(P<0.05). There were significant differences about the concentration of HBV DNAbetween CR and PR group in HBe Ag negative patients(P=0.001). The concentration of HBs Ag and HBe Ag at baseline in HBe Ag non-responder group was significantly higher than that in HBe Ag responder group(P<0.05).2Prediction of interferon treatment response by dynamically monitoring the concentration of HBV markers in HBe Ag-positive CHB patientsAfter 48 weeks treatment, 20 patients achieved CR and 41 patients achieved PR, including 2 patients with HBs Ag clearance. There were no significant differences at baseline in HBV DNA, Anti-HBc and ALT between the two groups. Baseline concentration of HBs Ag inthe CR group was significantly lower than that in the PR group(3.81±0.45 log10IU/m L vs. 4.12±0.57 log10IU/m L, respectively, P=0.037]; the concentration of HBs Ag at week 12 in the CR group was significantly lower than that in the PR group(2.94±1.37 log10IU/m L vs.3.60±0.59 log10IU/m L, respectively, P=0.001]; by ROC analysis, patients with baselineconcentration of HBs Ag<3.20log10IU/m L were most likely to obtain CR at the end of treatment(P=0.002). HBs Ag level declined rapidly during the first 24-week of treatment, which dropped significant slowly in the CR group than that in the PR group(3.7±0.47 log10IU/m L vs.4.08±0.62 log10IU/m L, respectively, P=0.017). Baseline concentration of HBe Ag in the CR group was significantly lower than that in PR group(431.34±577.37S/COvs.914.72±586.50S/CO, respectively, P=0.004]; through ROC analysis, patients with baseline concentration of HBe Ag<520 S/CO were most likely to obtain CR at the end of treatment(P=0.006). There was no difference of the concentration of Anti-HBc between the CR group and the PR group at each time points(P>0.05). HBV DNA level at week 24 in the CR group was significantly lower than that in the PR group(2.93±0.27 log10IU/m L vs.3.47±0.94 log10IU/m L, respectively, P=0.020]; paitents with HBV DNA less than 3350 IU/m Lat week 24 wouled achieve CR at the end(P=0.043). Multivariate analysis showed that combining baseline HBs Ag, baseline HBe Ag with HBV DNA at week 24 would effectively predict the outcome of IFN treatment.?Conclusion?1 Exploring the predictable markers of IFN therapy in patients with chronic hepatitis BSpecific SNPs in CYP27B1 and OAS2 locus, through individual and haplotype analysis, were associated with IFN therapy in CHB patients. The concentration of serum 1,25(OH)2VD3, HBs Ag, HBe Ag and HBV DNAwill effectively predict the efficacy of IFN.2Prediction of interferon treatment response by dynamically monitoring the concentration of HBV markers in HBe Ag-positive CHB patientsHBs Ag, HBe Ag and HBV DNA combined suggest important implications in predicting the 48-week PEG-IFN therapy efficacy in HBe Ag-positive CHB patients. Generally, the baseline and the 12 thweek HBs Ag, the baseline HBe Agand HBV DNA levels exhibited the greatest significance.