Association Between Host Factors And Antiviral Efficacy In Patiehts With Chronic Hepatitis B

BackgroundsChronic hepatitis B(CHB)，cirrhosis and hepatocellular carcinoma are the classic trilogyin patients with chronic hepatitis B virus (HBV) infection. Antiviral therapy was themain way of treatment in CHB patients, which including of interferon-a (IFN-a) andnucleoside/nucleotide analog. Virological clearance, delayed progression to cirrhosis orliver cancer, and increased survival are the long-term goals of antiviral therapy in CHBpatients. For a long time, liver function, HBV-5M and HBVDNA are the classis way ofevaluating state of CHB patients. More new indicators emerge with the progress ofscientific and etchnological.Our previous study has found that ESR1SNP wassigniifcantly associated with initial IFN-a virological response in CHB patients by usingPCR-RFLP analysis. No study has reported the association of ESR1SNP and IFN-atherapy. Thus, we investigated the possible association among gene SNPs，level of genemRNA expression and IFN-a therapy after enlarging sample size，prolonging follow-uptime. IFN-a and entecavir(ETV) have the highest efifcacy and lowest risk for resistanceand are recommended as ifrst-line treatment in nucleoside-naive CHB patients. We alsoinvestigated the association between ESR1SNPs and ETV therapy, with the respect ofdiscovering new predictor for ETV and IFN-a therapy. AimsIn this study, nucleoside-naive CHB patients with IFN-a or ETV therapy were enrolled，7SNPs (rs2077647，rs9340799，rs2234693，rs322354，rs2071430，rs3759755andrs3759756) and3gene mRNA expression (ESR1, MxA and eIF-2a gene) wereexamined, the association between antiviral eiffcacy and host baseline characters, genepolymorphism and gene mRNA expression were investigated.MethodsA total of248CHB patients were enrolled in this study, involving172HBeAg positivepatients with IFN-a therapy and76HBeAg positive/negative patients with ETV therapy.The eiffcacy of antiviral therapy was examined in all patients at4，12，48，72and96weeks. Genome DNA was abstracted by using QIAGEN DNA kit. Seven SNPs(ESRlgme rs2077647，rs9340799，rs2234693and rs322354; MxA gene rs2071430;eIF-2a gene rs3759755and rs3759756) were determined by using both polymerasechain reaction reaction-restriction fragment length polymorphism(PCR-RFLP) assayand time of lfight mass spectrometry method. SYBR Green RT-PCR was used todetermine PBMC gem(ESRl’ MxA and eIF2d) mRNA level before,during and afterIFN-a therapy in103HBeAg positive patients. Electrophoresis mobility shift (EMSA)was used to examined the ability of nuclear binding protein between rs3759756A and Galleles. All statistical analyses were performed by using SPSS software version13.0.Results1. By multiple analysis, our results suggest that baseline HBVDNA levels wereassociated with IFN-a rapidly and early virological response. Patients achievedrapidly virological response were prone to achieve early virological response.Baseline HBsAg level, HBeAg level^200S/C0, HBeAg level <lOS/CO at week12and patients with early virological response were also associated with IFN-a virological response at72weeks of therapy. Baseline HBVDNA levels may play animportant role in determining ETV eiffcacy after24and48weeks of treatment.2. We examined7SNPs(￡^7gene rs2077647，rs9340799，rs2234693and rs322354，MxA gene rs2071430，eIF-2a gene rs3759755and rs3759756) in172HBeAgpositive CHB patients with IFN-a therapy. Univariate analysis showed that earlyvirological response among patients carrying rs2077647TT，TC and CC genotypegroups were signiifcantly different(x~2=6.240，P=0.044); and patients carrying atleast one rs2077647T allele might possess higher non-virological response thanthose carrying CC genotype(TT+TC vs. CC, x~2=6.189，P=0.013). Though thesedifference disappeared after adjusting for baseline ALT level, baseline HBVDNAand RVR(OR:0.727，95%CI:0.279-1.889，P=0.512and OR:1.585，95%CI:0.277-9.060，P=0.604，respectively). By univariate analysis, virological responseat72weeks were signiifcant different among patients carrying rs2234693TT, TCand CC genotype(x~2=6.478，P=0.039)，and patients carrying at least oners2234693C allele were more likely possess virological response than those whocarrying CC genotype at72weeks of therapy(66.4%vs.46.4%，%2=6.258，P=0.012). Virological response at72weeks of therapy were signiifcant differentamong patients carrying rs2071430GG，GT and TT genotypes(x~2=8.079，P=0.018)，and patients carrying at least one rs2071430C allele were more likely toachieve virological response at72weeks of therapy(66.6%vs.37.0%，%2=5.899，P=0.015). Compared with AA genotype, patients carrying rs3759756AG genotypewere prone to achieve non-virological response at72weeks of therapy(77.3%vs.34.7%，x~2=14.498，尸<0.001). However, after adjusting for baseline HBsAg level,baseline HBsAg level, baseline HBeAg level, HBeAg at12weeks, RVR and EVR,only rs3759756SNP was associated with virological response at72weeks(OR=37.988，95%CI:1.665-86.6737，P=0.023)，and non-virological response was more likely happened in patients with rs3759756AG genotype rather than AA genotype.3.Besides，we also examined ESR12SNPs(rs2234693and rs9340799) in76CHBpatients with ETV therapy. Virological response at both48and96weeks of therapywere signiifcantly different among patients carrying rs2234693TT，TC and CCgenotype(57.1%vs.87.8%vs.58.3%, P=0.012and64.3%vs.96.7%vs.87.5%, P=0.018，respectively). By multiple analysis, patients carrying at least one rs2234693C allele were more likely to achieve virological response compared with TTgenotype at both48and96weeks of therapy (81.1%vs.57.1%，95%CI:1.026-14.785，P=0.046and94.7%vs.64.3%，95%CI:1.456-57.509, P=0.018，respectively).4.The mRNA level of gene ESR1, MxA and eIF-2a were evaluated in103HBeAgpositive CHB patients with IFN-a therapy. In female, the baseline￡57?7mRNAlevel in72weeks virological response group were signiifcantly higher thannon-virological response(9.2722土6.448vs.6.041土6.738，Z=-0.207，P=0.038)，and baseline PBMC￡57?7mRNA level was negatively correlated with serumbaseline level of HBVDNA(r=-0.750，P <0.001). In male, level of￡57?7mRNAincreased signiifcantly in virological response group than non-virological responseat72weeks of therapy (21.838±39.775vs.1.929±1.174，Z=-2.162，尸=0.031).5.Patients carrying rs3759756AG genotype shown higher eIF-2a mRNA level thanAA genotype. EMSA analysis shown that the ability of nuclear binding protein inrs3759756A allele was3fold than G allele, these may partly account for differenteIF-2a mRNA level between AA and AG genotype.Conclusions1. ESR1rs2234693and baseline HBVDNA levels may play an important role indetermining ETV virological response. 2.In nucleoside-naive CHB patients with IFN-a therapy, baseline HBVDNA levelswere associated with IFN-a rapidly and early virological response. Baseline HBsAglevel, HBeAg Ievel<200s/C0, EVR，HBeAg level<lOS/CO at week12and eIF-2ars3759756were also associated with virological response at72weeks of therapy. Infemale, baseline PBMC ESRlmRNA level may be a predictor of72weeksvirological response to IFN-a therapy, and was negatively correlation with serumlevel of baseline HBVDNA. In male, IFN-a markedly increased ESRlmRNAexpression in virological responders at72weeks of therapy.3.The level of eIF-2a mRNA was not associated with IFN-a virological response, butshown signiifcant difference between rs3759756AA and AG genotype. Thedifference of nuclear binding protein between A and G allele may be one of thereason.