共价闭合环状 DNA 转录调控不需要乙型肝炎病毒核心蛋白

StephenW

共价闭合环状 DNA 转录调控不需要乙型肝炎病毒核心蛋白
钟有泉#1 2、吴传建#1、徐再超1、严腾1、李兆1、赵开涛1、王晶晶1、文王3、琼占1、朱成良4、陈新文5、梁凯伟3 , 程晓明 1 6 7 , 夏雨辰 1
隶属关系
隶属关系

    1
    武汉大学太康医学院太康生命医学中心医学病毒学研究所病毒学国家重点实验室、过敏与免疫学湖北省重点实验室，武汉大学grid.49470.3e。
    2
    【作者单位】： 广西壮族自治区人民医院检验科;
    3
    武汉大学太康医学院太康生命与医学中心病理生理学教研室.49470.3e,武汉
    4
    武汉大学人民医院临床检验科，49470.3e，武汉，中国。
    5
    中国科学院武汉病毒研究所病毒学国家重点实验室，武汉，中国。
    6
    武汉大学网格.49470.3e 武汉大学中南医院病理与分子诊断中心，武汉，中国。
    7
    湖北省临床中心和大肠疾病重点实验室，武汉，中国。

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同等贡献。

    PMID：36226986 DOI：10.1128/jvi.01362-22

抽象的

乙型肝炎病毒 (HBV) 感染是全球主要的健康负担，目前尚无治愈方法。 HBV共价闭合环状DNA（cccDNA）的持续存在是抗病毒治疗的主要障碍。 HBV核心蛋白（HBc）已成为一种有前途的抗病毒靶点，因为它在病毒生命周期的关键步骤中发挥着重要作用。然而，HBc 是否可以调节 HBV cccDNA 转录仍存在争议。在这项研究中，不同的方法被用来解决这个问题。在合成的 HBV cccDNA 和 HBVcircle 转染试验中，缺乏 HBc 对肝癌细胞系和原代人肝细胞中 HBV RNA 的转录以及 HBV 表面抗原 (HBsAg) 的产生没有影响。 HBc 的重组不会改变 cccDNA 衍生的 HBV 标志物的表达。从含有 cccDNA 的体内小鼠模型中获得了类似的结果。染色质免疫沉淀 (ChIP) 或 ChIP 测序分析揭示了与野生型 cccDNA 相似的 HBc 缺陷型 cccDNA 染色质的转录调控。此外，用衣壳组装调节剂 (CAM) 治疗可显着减少细胞外 HBV DNA，但不能改变病毒 RNA 和 HBsAg。我们的结果表明，HBc 既不影响 cccDNA 的组蛋白修饰和转录因子结合，也不直接影响 cccDNA 转录。尽管 CAM 可以减少 HBc 与 cccDNA 的结合，但它们不会抑制 cccDNA 转录活性。因此，不应期望靶向衣壳或 HBc 的治疗剂充分减少 cccDNA 转录。重要性 乙型肝炎病毒 (HBV) 核心蛋白 (HBc) 已成为一种有前途的抗病毒靶点。然而，HBc 是否可以调节 HBV 共价闭合环状 DNA (cccDNA) 的转录仍然难以捉摸。本研究表明HBc对cccDNA的表观遗传调控没有影响，不参与cccDNA的转录。鉴于 HBc 对于 cccDNA 转录是可有可无的，因此需要新的 cccDNA 靶向治疗来治愈 HBV。

关键词：衣壳抑制剂； cccDNA；核心蛋白；乙型肝炎病毒;转录调控。

StephenW

Hepatitis B Virus Core Protein Is Not Required for Covalently Closed Circular DNA Transcriptional Regulation
Youquan Zhong #  1   2 , Chuanjian Wu #  1 , Zaichao Xu  1 , Yan Teng  1 , Li Zhao  1 , Kaitao Zhao  1 , Jingjing Wang  1 , Wen Wang  3 , Qiong Zhan  1 , Chengliang Zhu  4 , Xinwen Chen  5 , Kaiwei Liang  3 , Xiaoming Cheng  1   6   7 , Yuchen Xia  1
Affiliations
Affiliations

    1
    State Key Laboratory of Virology and Hubei Province Key Laboratory of Allergy and Immunology, Institute of Medical Virology, TaiKang Center for Life and Medical Sciences, TaiKang Medical School, Wuhan Universitygrid.49470.3e, Wuhan, China.
    2
    Department of Laboratory Medicine, The People's Hospital of Guangxi Zhuang Autonomous Region, Nanning, China.
    3
    Department of Pathophysiology, TaiKang Center for Life and Medical Sciences, TaiKang Medical School, Wuhan Universitygrid.49470.3e, Wuhan, China.
    4
    Department of Clinical Laboratory, Renmin Hospital of Wuhan Universitygrid.49470.3e, Wuhan, China.
    5
    State Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, China.
    6
    Wuhan Universitygrid.49470.3e Center for Pathology and Molecular Diagnostics, Zhongnan Hospital of Wuhan University, Wuhan, China.
    7
    Hubei Clinical Center and Key Laboratory of Intestinal and Colorectal Diseases, Wuhan, China.

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Contributed equally.

    PMID: 36226986 DOI: 10.1128/jvi.01362-22

Abstract

Hepatitis B virus (HBV) infection is a major health burden worldwide, and currently there is no cure. The persistence of HBV covalently closed circular DNA (cccDNA) is the major obstacle for antiviral trement. HBV core protein (HBc) has emerged as a promising antiviral target, as it plays important roles in critical steps of the viral life cycle. However, whether HBc could regulate HBV cccDNA transcription remains under debate. In this study, different approaches were used to address this question. In synthesized HBV cccDNA and HBVcircle transfection assays, lack of HBc showed no effect on transcription of HBV RNA as well as HBV surface antigen (HBsAg) production in a hepatoma cell line and primary human hepatocytes. Reconstitution of HBc did not alter the expression of cccDNA-derived HBV markers. Similar results were obtained from an in vivo mouse model harboring cccDNA. Chromatin immunoprecipitation (ChIP) or ChIP sequencing assays revealed transcription regulation of HBc-deficient cccDNA chromatin similar to that of wild-type cccDNA. Furthermore, treatment with capsid assembly modulators (CAMs) dramatically reduced extracellular HBV DNA but could not alter viral RNA and HBsAg. Our results demonstrate that HBc neither affects histone modifications and transcription factor binding of cccDNA nor directly influences cccDNA transcription. Although CAMs could reduce HBc binding to cccDNA, they do not suppress cccDNA transcriptional activity. Thus, therapeutics targeting capsid or HBc should not be expected to sufficiently reduce cccDNA transcription. IMPORTANCE Hepatitis B virus (HBV) core protein (HBc) has emerged as a promising antiviral target. However, whether HBc can regulate HBV covalently closed circular DNA (cccDNA) transcription remains elusive. This study illustrated that HBc has no effect on epigenetic regulation of cccDNA, and it does not participate in cccDNA transcription. Given that HBc is dispensable for cccDNA transcription, novel cccDNA-targeting therapeutics are needed for an HBV cure.

Keywords: capsid inhibitor; cccDNA; core protein; hepatitis B virus; transcriptional regulation.