NQO1调控HBx蛋白稳定性对乙肝病毒转录和复制的重要作用

Hepatitis B virus (HBV) infection remains a major health problem worldwide and no treatment can completely eliminate HBV replication in hepatocytes in all patients. Therefore, characterization of HBV replication, especially cccDNA transcription, will provide valuable information for the development of anti-HBV therapeutics. Based on RNA-Sequencing analysis of liver tissue from CHB patients and inactive carrier, we found that NQO1 was upregualted in liver from CHB patients. This upregulation was further confirmed in another 20 liver tissues from CHB patients. Furthermore, NQO1 overexpression stabilized cellular HBx protein and enhanced total HBV RNAs and 3.5kb RNA, suggesting NQO1 might regulated cccDNA transcription though stabilizing HBx protein. Bases on above findings, this study will further investigate the mechanism by which NQO1 stabilized HBx in HBV infection cell model HepG2-NTCP and PHH. We will also investigate the important role of NQO1/HBx in cccDNA transcription. Finally, we will examined the antiviral activity of NQO1 inhibitor on HBV replication on chronic hepatitis mouse model. This study will provide deeply understanding of HBx function and HBV replication and can provide new strategy for reduction HBV infection rate and identification a novel target to treatment of hepatitis B.

乙型肝炎病毒(HBV)感染呈世界范围内广泛流行，但现有药物治疗乙肝尚存在不少问题；深入研究HBV复制调控机制，将极大促进抗病毒治疗新手段的发展。通过肝组织RNA-Seq分析，发现宿主因子醌氧化还原酶1(NQO1)与HBV复制相关。临床样本分析证实NQO1mRNA在慢乙肝病人肝组织中显著增加；细胞模型研究发现NQO1可稳定HBx蛋白，并显著增加HBV总RNA和3.5kb-RNA的表达，提示NQO1稳定HBx的这个功能可能对HBV复制，尤其cccDNA转录环节发挥重要作用。基于上述发现，本项目将在HBV感染模型HepG2-NTCP和PHH中，清晰解析NQO1稳定HBx蛋白的分子机制，阐明NQO1稳定HBx对HBV转录和复制过程的调控机制，在动物模型中评估NQO1抑制剂降解HBx和抗病毒效能。本项目的开展将加深对HBx蛋白功能的了解，为探索治疗乙型肝炎新的靶点提供理论依据。

乙型肝炎病毒(HBV)感染呈世界范围内广泛流行，但现有药物不能有效清除病毒转录复制模板cccDNA。因此，深入研究HBV复制调控机制，有利于寻找有效抑制cccDNA转录靶点。通过肝组织RNA-Seq分析，发现宿主因子醌氧化还原酶1(NQO1)与HBV复制相关。基于HBV感染模型HepG2-NTCP和PHH中的实验分析证实NQO1mRNA在HBV感染细胞中显著增加。机制研究发现NQO1可稳定HBx蛋白，并显著增加HBV总RNA和3.5kb-RNA的表达，提示NQO1稳定HBx的这个功能可能对HBV复制，尤其cccDNA转录环节发挥重要作用。进一步研究表明，NQO1通过稳定HBx，进而参与cccDNA结合组蛋白的转录修饰调控，最终促进cccDNA转录。最后，我们在动物模型中评估了NQO1抑制剂降解HBx和抗病毒效能。本项目的开展将加深对HBx蛋白功能的了解，为探索治疗乙型肝炎新的靶点提供理论依据。