从IFN信号通路寻找影响CHB患者IFN-α疗效差异的关键基因并证实其功能

Treatment response of interferon alpha (IFN-α) for chronic hepatitis B (CHB) patients has significant individual difference, the mechanism of which remains unknown. Currently, evaluation of treatment response to IFN in CHB is mainly according to the change of serum markers in the course of treatment. However, we cannot individually predict the curative effect of IFN before starting treatment..In present study, we have found 3 genes which are related to treatment difference of IFN. Case-control study is employed in the study. The project intends to screen potential key genes affecting IFN-α treatment from the 17 genes in the IFN signaling pathway through the Hapmap database. Then, SNP typing was performed by iMLDR technology, relationship between SNPs and IFN response in patients with CHB is analyzed. Further, functional study of SNPs from the gene, protein and cell level is performed. Finally, we explore the diagnostic value of single or multiple SNPs to construct artificial neural network model to predict the treatment efficacy of IFN for the Chinese Han population..In theory, the research is expected to find diagnostic markers of SNP associated with treatment differences of IFN in CHB, and to reveal the potential molecular mechanism. In practice, the study is expected to provide experimental basis for the implementation of individualized treatment to avoid the side effects and the economic burden for those without response to IFN. Therefore, the project has important theoretical and practical significance.

慢性乙型肝炎（CHB）患者的α干扰素（IFN-α）疗效存在显著个体差异，其机制尚不明了。目前主要根据治疗中血清学标志物变化判断患者对IFN的应答，但治疗前无法个性化预测对IFN的应答。.前期我们已发现IFN受体基因等3个基因多态性与疗效有关。本研究拟采用病例-对照的方法，从IFN信号通路的17个基因入手，通过Hapmap数据库筛选出可能影响IFN-α应答的SNPs，用iMLDR技术进行SNP分型，分析其与CHB患者IFN应答的关系；从基因、蛋白和细胞水平研究应答相关的SNP的功能；探讨单个/多个SNP联合的诊断价值，构建适合中国汉族人群的人工神经网络模型以预测IFN疗效。.本研究理论上有望找到与CHB患者IFN疗效有关的SNP诊断标志物，揭示其差异的分子遗传学机制；实践中有望为实施个体化诊疗、避免无应答者使用IFN带来的毒副作用和经济负担提供依据，有重要的理论和实际意义。

干扰素α（IFNα）治疗慢性乙型肝炎（CHB）患者的疗效存在个体差异，本课题筛选与IFNα疗效相关的SNP位点，并探讨其影响疗效的机制。我们通过ASA芯片和Massarray技术，找到P<0.05的位点23672个，P<0.01的位点4381个（基因1265个），P<10-5的位点51个（基因20个）。.筛选出差异SNPs后，我们通过GeneCards和文献查阅P<10-5位点对应的基因功能和注释，从中挑选14个基因/蛋白进行定量检测。结果发现，ZNF350基因mRNA在应答不佳（SR）组中的表达量显著高于完全应答（CR）组；载脂蛋白B（APOB）的蛋白表达量在CR组的表达量显著低于SR组中。通过进一步统计分析，我们发现APOB rs1367117、APOB rs10199768、ZNF350 rs2278420、ZNF350 rs6509607、TRIM22 rs10838543、TRIM15 rs2074477的基因型与IFNα疗效有关。在功能学研究方面，我们培养HBV感染者PBMC并用IFNα刺激后发现，rs10199768 AC型感染者的STAT1和CXCL10的mRNA表达水平较CC型高；单倍型分析结果显示，rs1367117A/rs10199768 A感染者的抗病毒分子STAT1、CXCL10和ISG15的表达水平较rs1367117G/rs10199768C高，A等位基因与完全应答有关；rs2278420 AA型感染者的IFNα通路负调控基因SOCS1、SOCS3和PIAS1表达水平较AG型高；rs6509607 AA型感染者的负调控基因PIAS1、PTPN6的表达水平较AG型高，A等位基因与应答不佳有关；TRIM22 rs10838543 CC型感染者抗病毒蛋白IFIT3的表达水平较TT型高。另外在Hep AD38细胞实验中，用IFNα刺激后，含TRIM22 rs10838543 C等位基因的抗病毒蛋白（PKR、IFIT3、OAS1、MxA）的表达量显著高于T等位基因，T等位基因与应答不佳有关。.综上，本研究找到与CHB患者IFNα疗效有关的SNP诊断标志物并探讨了其影响疗效的机制；本研究有望为实施个体化诊疗、避免无应答者使用IFN带来的毒副作用和经济负担提供依据，有重要的理论和实际意义。