hESC-derived hepatocytes 中抗HBV干扰素反应模式及关键ISGs 的功能分析

由于缺乏能真实反映病毒与宿主之间相互作用的HBV研究模型，HBV引发肝细胞产生干扰素反应模式及关键抗病毒干扰素刺激基因（ISGs）迄今仍未明了。申请人在前期工作中，稳定定向分化人胚胎干细胞为肝细胞 (hESC-Heps)，成功建立肝炎性病毒感染研究模型，该模型更加真实的呈现了人原代肝细胞的天然免疫反应。在此基础上，本项目将进一步1）稳定定向分化人胚胎干细胞获得肝细胞hESC-Heps，考察其不同分化阶段对外源性干扰素IFNa/β/λ反应模式特点；2）比较HBV在未成熟的和成熟的hESC-Heps中引发的干扰素反应模式和ISGs特点；3）运用RNA干扰、表达谱分析等技术确定关键ISG基因的抗病毒功能及其抗病毒分子机制。项目预期通过该新型细胞模型，揭示HBV引发宿主细胞的干扰素反应模式特点及其同肝细胞分化成熟的关联性，获得干扰素通路中关键的具有抗病毒功能的ISGs，为乙肝治疗提供重要参考依据。

Up to now, pattern of anti-HBV response induced by the interferons and their anti-HBV mechanisms have not yet been fully clarified due to the lack of HBV model that can really mimic virus-host interaction. In our previous work, we have successfully differentiated human embryonic stem cell into hepatocytes (hESC-Heps) and modeled hepatitis viral infection. Furthermore, this novel cell model can reflect innate immune response of human primary hepatocytes authentically (Zhou XL, et al. Stem Cell Reports, 2014). This project is composed of 3 major parts including 1) stably differentiate hESC into hESC-Heps and study the patterns of interferon response induced by the exogenous IFN a/β/λ, respectively; 2) compare the patterns of interferon response and interferon-stimulated genes (ISGs) induced by HBV in immature and mature hESC-Heps, respectively; 3) apply the technologies of RNA interference, analysis of mRNA expression profile, gene over expression etc to confirm the key ISGs with anti-HBV function as well as the mechanisms of these ISGs. The project is aimed at disclosing the characteristics of the pattern of interferon response induced by HBV in hepatocytes, getting the key ISGs with anti-HBV function, and therefore providing the important data for Hepatitis B therapy as well as the development of novel anti-HBV drugs.