慢性乙型肝炎难以治愈，研发更有效的抗HBV疗法势在必行，因此需要探索新的抗病毒靶点。激酶参与多种病毒感染过程，但在HBV感染中的作用尚未明确。我们在预实验中应用激酶小干扰RNA库筛选HBV感染关键激酶，发现敲低激酶AAK1表达显著降低HBV感染。进一步发现，激酶AAK1通过磷酸化宿主蛋白AP-2的µ亚单位，激活AP-2与病毒蛋白HBsAg结合。激酶AAK1抑制剂7745在体外表现出抗HBV活性。据此，我们首次提出“激酶AAK1通过磷酸化AP-2调控HBV感染，可作为抗HBV靶点”的假说。本项目将应用HBV细胞和动物模型进一步研究以AAK1为代表的激酶对HBV入胞、复制、组装、释放、传播、cccDNA形成的影响，阐明激酶AAK1通过磷酸化AP-2，激活AP-2与病毒蛋白在细胞内共转运，调控HBV感染的分子机制，并探索以AAK1等激酶为靶点的抗HBV新策略，为研发新型抗HBV疗法提供理论依据。

The low cure rate for chronic hepatitis B achieved with the current treatments makes it necessary to explore new antiviral targets for development of more effective antiviral therapies. Kinases participate in the life cycle of multiple viruses and therefore become promising targets for antiviral therapies. However, the role of kinases in HBV infection remains unknown. In our pilot experiment, we screened for key kinases for hepatitis B virus (HBV) infection using a kinase siRNA library and found that knockdown of AAK1 significantly reduced HBV infection. Furthermore, we found that AAK1 phosphorylated µ subunit of host protein AP-2 to activate binding of AP-2 and viral protein HBsAg. 7745, a small molecule targeting AAK1, exhibited anti-HBV activity in the HBV cell culture model. Based on these results, we hypothesized for the first time that AAK1 kinase regulates HBV infection through AP-2 phosphorylation and therefore could be a promising anti-HBV target. In this study, we will use HBV cell culture and animal models to further investigate the impact of AAK1 and other kinases on HBV entry, replication, assembly, release and cccDNA formation, and elucidate the molecular mechanism of AAK1 kinase in regulating HBV infection through phosphorylation of AP-2 to activate intracellular co-trafficking of AP-2 and viral protein. Finally, we will explore novel anti-HBV strategies of using HBV-associated key kinases as targets to inhibit HBV infection. Our study will provide proof of concept for the development of novel anti-HBV therapies.