HBV pgRNA病毒样颗粒侵入肝细胞并形成持续感染能力的研究

申请人近年专注HBV RNA相关研究并在本领域权威期刊发表相关论文4篇。近期研究证实血清HBV RNA在预测乙肝抗病毒治疗应答方面有重要价值，血清中HBV RNA以pgRNA为主且部分以病毒样颗粒的形式存在。我们前期研究发现HBV pgRNA病毒样颗粒具有与HBV DNA病毒颗粒相同的结构和分泌机制并建立了分离纯化pgRNA病毒样颗粒的方法。本研究拟通过HepG2-NTCP细胞和人原代肝细胞，证明pgRNA病毒样颗粒能够内化进入肝细胞并形成cccDNA，进而评估细胞内外多种复制指标的动态变化，证明pgRNA病毒样颗粒感染肝细胞后能够合成并分泌新的病毒颗粒，最后在人源化肝脏小鼠模型中证明pgRNA病毒样颗粒的感染能力。研究将阐明HBV pgRNA病毒样颗粒是否具备感染肝细胞并形成持续感染的能力，有助于完善对HBV复制周期的认识并为乙肝抗病毒药物的研发提供新靶点，扩大血清HBV RNA临床应用

Serum HBV RNA plays an important role in predicting responses after antiviral treatment. It has also been verified that HBV pgRNA accounts for the majority of serum HBV RNA, some of which existed as virion-like particles. The applicant has published 4 papers related to HBV RNA in peer-reviewed journals. Our preliminary data showed that we have established a method for isolating and purifying pgRNA virion-like particles. We found that the structure and secretion mechanism of HBV pgRNA virion like particles is identical to HBV DNA virion. Herein, this project intends to determine the capability of HBV pgRNA virion like particles in the infection of hepatocyte and establishment of persistent infection by in vitro and in vivo experiments. With the cell culture model of HepG2-NTCP cells and human primary hepatocytes, we plan to demonstrate the internalization ability of pgRNA virion like particles and the following intrahepatic cccDNA formation, then evaluate the dynamics of different markers reflecting viral replication, to prove the ability of secreting new virions after infection of hepatocyte by pgRNA virion like particles. Furthermore, a mouse model with humanized liver will be used to confirm the infection capability of pgRNA virion like particle. Hopefully, our study could contribute to a better understanding of HBV life cycle, shed some light on new targets for anti-HBV drug development and broaden the area for clinical application of serum HBV RNA.