SART1通过调节MX1剪接和lncRNA-IFI6表达影响HBV及HCV复制的机制研究

Hepatitis B Virus （HBV）and Hepatitis C virus (HCV）are major causes of chronic liver hepatitis diseases. Nucleoside analogues and direct antiviral agents (DAAs) drugs have good efficacy in the treatment of HBV and HCV infections. However, there are still frequent viral recurrences and high treatment costs. The molecular mechanism by which interferon-α anti–HCV, -HBV effects are not well defined. We previously identified SART1 (squamous cell carcinoma antigen recognized by T-cells 1), a host protein involved in RNA splicing and pre-mRNA processes, as a regulator of IFN antiviral effects. We performed RNA-Seq and siRNA knockdown in Huh7.5.1 cells with or without IFN treatment. We identified that SART1 knockdown reduced classical ISG mRNA expression of MX1 and changed alternative mRNA splicing that contribute to their antiviral effects. In addition, further data mining from our RNA-seq data revealed a cluster of lncRNA (long non-coding RNA) potentially involved in IFN’s antiviral regulation. In this proposal we plan to explore the detailed mechanism by which HCV, HBV replication regulated by antiviral gene MX1 variants and the selected lncRNA IFI6. We will explore these mechanisms in culture cells including Huh7.5.1, JFH1 HCV-infected Huh7.5.1, HepG2, HBV AD38 cells. We plan to verify the novel findings in primary human hepatocytes (PHHs). We will also monitor these MX1 variants and lncRNA IFI6 expression in human PBMC, liver biopsy to assess their associations with sustained virological response (SVR) or non-SVR to interferon-α treatment. The findings in this grant proposal will provide new insight into the RNA splicing mechanisms and novel MX1 gene variants for possible clinical application for HCV, HBV therapy. Our study will help to define the important biological mechanism and to develop new clinical diagnostic test and marker in the translational settings in HCV, HBV therapy to improve patient care.

乙肝病毒HBV和丙肝病毒HCV感染是导致严重肝脏疾病的主要原因。尽管核苷类似物及DAAs药物在治疗病毒感染方面具有很好的疗效，但易复发及价格昂贵。我们前期发现剪接因子SART1调节MX1的可变剪接和长链非编码lncRNA IFI6的表达，影响干扰素的抗病毒作用。本项目拟深入探索MX1剪接变异体及lncRNA IFI6如何调控HBV和HCV的复制。我们将高表达MX1剪接异构体，采用合成MX1剪接异构体短肽，检测其对HBV及HCV复制的影响. 探索其作为小分子抗病毒药物的可能性.另一方面，我们拟将在细胞中高表达或者gRNA敲除lncRNA IFI6，考察其对病毒复制的调节及可能的作用方式。在细胞株实验的基础上,计划在人类原代肝细胞中验证这些新发现。 我们也将从临床水平验证这些基因和其剪接变异体与干扰素治疗效果的相关性。研究结果将为新的RNA剪接理论提供依据并发现可能应用于临床抗病毒潜在靶点。

乙肝病毒(HBV)和丙肝病毒(HCV)感染是导致严重肝脏疾病的主要原因。尽管核苷类似物及DAAs药物在治疗病毒感染方面具有很好的疗效，但HBV易复发及HCV药物价格昂贵。我们前期发现剪接因子SART1调节MX1的可变剪接和长链非编码lncRNA IFI6的表达，影响干扰素的抗病毒作用。本项目研究了MX1剪接变异体及lncRNA IFI6如何调控HBV和HCV的复制。研究结果显示，病毒感染和干扰素显著调节MX1剪接体的表达；HBV感染抑制MX1的表达，MX1 V9-V13显著抑制HBV的复制。IFN诱导的lncRNA-IFI6通过调节IFI6的启动子活性和组蛋白修饰，影响HCV的复制。此外，我们证实了患者肝活检中相关基因表达（IFI6，MX1）的抑制与HCV，HBV感染之间的关联。研究结果将为新的RNA剪接理论提供依据并发现可能应用于临床抗病毒潜在靶点，同时有助于了解重要的生物学机制，为开发新的临床诊断标志物和可能应用于临床抗病毒的潜在靶点，改善HCV、HBV患者的治疗效果提供依据。

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