HDAC6调控HBV诱导自噬及影响HBV自我复制的作用与机制研究

本课题组的前期研究发现，干扰素(IFN)诱导的TRIM22分子在抗HBV固有免疫中发挥重要作用； 组蛋白去乙酰化酶（HDAC）6在IFN调控TRIM22的表达中起到重要作用。在HepG2.2.15细胞中抑制HDAC6的活性可明显促进HBV复制，但意外发现其作用不依赖于IFN及TRIM22等干扰素刺激基因的表达，而可能与HDAC6对自噬的调控密切相关。本研究拟通过过表达、RNA干扰及特异性化学阻断剂等方法确证HDAC6通过影响细胞自噬进而调控HBV复制的作用；通过免疫共沉淀确定HDAC6的去乙酰化底物及相互作用蛋白，关注HDAC6介导的乙酰化修饰在细胞自噬及HBV复制中的作用；并将探究HDAC6可否通过影响自噬相关基因的表达、自噬相关信号通路以及内质网应急等来调控自噬和HBV复制； 另外将重点探究HDAC6 对HBV诱导的自噬体与溶酶体融合的影响；为基于自噬的抗HBV作用提供新的策略及靶标。

Our previous investigation demonstrated that IFN-inducible TRIM22 played an important role in innate antiviral immunity, and Histone Deacetylase (HDAC) 6 is required for TRIM22 induction by IFN.It was found,surprisingly,that inhibition of HDAC6 significantly enhanced HBV replication in HepG2.2.15 cells, which didn't involve the suppresion of IFN,TRIM22 and other IFN-stimulate genes, but might depend on its effect on autophagy induction.In present study, we will futher demonstrate that HDAC6 regulate HBV replication via its effect on autophagy induction by gain - and loss-of-fuction analysis; immunoprecipitaion analysis will be used to dectect acetylated substrate upon HDAC6 inbition, and to detect the proteins that may interact with HDAC6, with the aim to deciper the HDAC6-mediated acetylated posttranscriptional modification on autophagy and HBV replication; we also will investigate whether HDAC6 exert its effct on autophagy and HBV replication via regulating the expression of autophagy-related genes, autophagy-related signaling pathways or ER stress; particular attention will also be put on the effect of HDAC6 on the fusion of HBV-induced autophagesome with lysosome.This study may help to design new anti-HBV strategy.