Regulatory function of interferon‐inducible 44‐like for hepatitis B virus covalently closed circular DNA in primary human hepatocytes
Regulatory function of interferon‐inducible 44‐like for hepatitis B virus covalently closed circular DNA in primary human hepatocytes
复制标题
干扰素诱导型44样蛋白对原代人肝细胞中乙型肝炎病毒共价闭合环状DNA的调节功能
DOI：
10.1111/hepr.13722
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发表时间：
2021
影响因子：
4.2
通讯作者：
Nakamoto Yasunari
中科院分区：
文献类型：
--
作者：
Nosaka Takuto;Naito Tatsushi;Murata Yosuke;Matsuda Hidetaka;Ohtani Masahiro;Hiramatsu Katsushi;Nishizawa Tsutomu;Okamoto Hiroaki;Nakamoto Yasunari
目的治疗B型肝炎病毒（HBV）感染需要消除共价闭合环状DNA（cccDNA）。干扰素（IFN）-γ具有非细胞溶解性抗病毒潜力;然而，无法消除cccDNA。为了增强调节作用，我们使用显示各种转录水平的体外HBV感染系统，全面分析了与cccDNA扩增和IFN-γ和IFN-α效应相关的宿主因素。方法使用携带基本核心启动子突变A1762 T/G1764 A和/或前核心突变G1896 A的基因组质粒，用HBV感染原代人肝细胞，并用IFN-γ和IFN-α处理。涉及微阵列和小干扰RNA分析的全面和功能研究揭示了与cccDNA regulation. ResultsHBV感染系统复制HBV生命周期相关的宿主因素，并表现出不同的繁殖水平。微阵列分析显示53个基因与cccDNA水平相关。在53个基因中，IFN-γ和IFN-α显著上调IFN-诱导蛋白44样（IFI 44 L）的表达。IFI 44 L的抗HBV作用是通过抑制核因子-κB和信号转导子和转录激活子1途径的激活而发挥的，与IFN-γ或IFN-α无关。结论使用体外HBV感染系统，鉴定了与cccDNA扩增相关的IFN-诱导分子IFI 44 L。这些结果表明，通过控制新的宿主因子IFI 44 L来调节HBV cccDNA的创新分子策略。
AimCuring hepatitis B virus (HBV) infection requires elimination of covalently closed circular DNA (cccDNA). Interferon (IFN)‐γ has noncytolytic antiviral potential; however, elimination of cccDNA could not be achieved. To enhance the regulatory effect, we comprehensively analyzed the host factors associated with cccDNA amplification and IFN‐γ and IFN‐α effects using an in vitro HBV infection system showing various transcription levels.MethodsPrimary human hepatocytes were infected with HBV using genomic plasmids carrying the basic core promoter mutation A1762T/G1764A and/or the precore mutation G1896A and treated with IFN‐γ and IFN‐α. Comprehensive and functional studies involving microarray and small interfering RNA analysis revealed the host factors related to cccDNA regulation.ResultsThe HBV infection system reproduced the HBV life cycle and showed various propagation levels. Microarray analysis revealed 53 genes correlated with the cccDNA levels. Of the 53 genes, expression of IFN‐induced protein 44‐like (IFI44L) was significantly upregulated by IFN‐γ and IFN‐α. The anti‐HBV effect of IFI44L is exerted regardless of IFN‐γ or IFN‐α by inhibiting the activation of nuclear factor‐κB and signal transducer and activator of transcription 1 pathways.ConclusionsUsing the in vitro HBV infection system, an IFN‐inducible molecule, IFI44L, associated with cccDNA amplification, was identified. These results suggest an innovative molecular strategy for the regulation of HBV cccDNA by controlling a novel host factor, IFI44L.