Polo样激酶1是乙肝病毒复制的前病毒宿主因子

StephenW

Polo-like-kinase 1 is a proviral host factor for hepatitis B virus replication

    Ahmed Diab1,2,3,†, Adrien Foca1,2,‡,*, Floriane Fusil2,4, Thomas Lahlali1,2, Pascal Jalaguier1,2, Fouzia Amirache4, Lia N'Guyen1,2, Nathalie Isorce1,2, François-Loïc Cosset2,4, Fabien Zoulim1,2,5,6, Ourania Andrisani3 andDavid Durantel1,2,6,*

Version of Record online: 6 NOV 2017

DOI: 10.1002/hep.29236

© 2017 by the American Association for the Study of Liver Diseases.

Issue
Hepatology

Volume 66, Issue 6, pages 1750–1765, December 2017
Article has an altmetric score of 3

    1    INSERM U1052, Cancer Research Center of Lyon, Lyon, France
    2    University of Lyon, Université Claude-Bernard, UMR_S1052, UCBL, Lyon, France
    3    Department of Basic Medical Sciences and Purdue Center for Cancer Research, Purdue University, West Lafayette, IN
    4    CIRI–International Center for Infectiology Research, Team EVIR, INSERM, U1111, Université Claude Bernard Lyon 1, CNRS, UMR5308, Ecole Normale Supérieure de Lyon, Univirsity of Lyon, Lyon, France
    5    Hepato-Gastroenterogy Unit, Croix-Rousse Hospital, Hospices Civils de Lyon, Lyon, France
    6    Labex DEVweCAN, Lyon, France

    ‡    Potential conflict of interest: Nothing to report.
    †    These authors contributed equally to this work.

Email: Adrien Foca ([email protected]), David Durantel ([email protected])

*ADDRESS CORRESPONDENCE AND REPRINT REQUESTS TO:
David Durantel, Ph.D.
INSERM U1052
Cancer Research Center of Lyon
Lyon 69008, France
E-mail: [email protected]
Tel: +33-472-681-970/+33-472-681-959
or
Ourania Andrisani, Ph.D.
Department of Basic Medical Sciences
Purdue University
West Lafayette, IN 47907 USA
E-mail: [email protected]
Tel: +1-765-494-8131

    This work was supported by grants from ANRS (French national agency for research on AIDS and viral hepatitis; several grants from CSS4) and by INSERM core grants to F.L.C., F.Z., and D.D. This work was also supported by the DEVweCAN LABEX (ANR-10-LABX-0061) of the Université de Lyon within the program Investissements d'Avenir (ANR-11-IDEX-0007) operated by the French National Research Agency to F.Z. and D.D. and National Institutes of Health grant (DK044533-19) to O.A. The work in the laboratory of FL Cosset was further supported by the European Research Council (ERC-2008-AdG-233130-HEPCENT) and the LabEx Ecofect (ANR-11-LABX-0048). A.D. received a Chateaubriand Fellowship from the French Embassy in Washington, DC.



Chronic hepatitis B virus (HBV) infection is a major risk factor for hepatocellular carcinoma (HCC) and current treatments for chronic hepatitis B and HCC are suboptimal. Herein, we identified cellular serine/threonine Polo-like-kinase 1 (PLK1) as a positive effector of HBV replication. The aim of this study was to demonstrate the proviral role of PLK1 in HBV biosynthesis and validate PLK1 inhibition a potential antiviral strategy. To this end, we employed physiologically relevant HBV infection models of primary human hepatocytes (PHHs) and differentiated HepaRG cells in conjunction with pharmacologic PLK1 inhibitors, small interfering RNA (siRNA)-mediated knockdown, and overexpression of constitutively active PLK1 (PLK1CA). In addition, a humanized liver Fah−/−/Rag2−/−/Il2rg−/− (FRG) mouse model was used to determine the antiviral effect of PLK1 inhibitor BI-2536 on HBV infection in vivo. Finally, in vitro PLK1 kinase assays and site-directed mutagenesis were employed to demonstrate that HBV core protein (HBc) is a PLK1 substrate. We demonstrated that HBV infection activated cellular PLK1 in PHHs and differentiated HepaRG cells. PLK1 inhibition by BI-2536 or siRNA-mediated knockdown suppressed HBV DNA biosynthesis, whereas overexpression of PLK1CA increased it, suggesting that the PLK1 effects on viral biosynthesis are specific and that PLK1 is a proviral cellular factor. Significantly, BI-2536 administration to HBV-infected humanized liver FRG mice strongly inhibited HBV infection, validating PLK1 as an antiviral target in vivo. The proviral action of PLK1 is associated with the biogenesis of the nucleocapsid, as BI-2536 leads to its decreased intracellular formation/accumulation. In this respect, our studies identified HBc as a PLK1 substrate in vitro, and mapped PLK1 phosphorylation sites on this protein. Conclusion: PLK1 is a proviral host factor that could be envisaged as a target for combined antiviral and antitumoral strategies against HBV infection and HBV-mediated carcinogenesis. (Hepatology 2017;66:1750–1765)

StephenW

Polo样激酶1是乙肝病毒复制的前病毒宿主因子

    Ahmed Diab1,2,3，Adrien Foca1,2，*，Floriane Fusil2,4，Thomas Lahlali1,2，Pascal Jalaguier1,2，Fouzia Amirache4，Lia N'Guyen1,2，Nathalie Isorce1,2，François- LoïcCosset2,4，Fabien Zoulim1,2,5,6，Ourania Andrisani3和David Durantel1,2,6，*

在线记录版本：2017年11月6日

DOI：10.1002 / hep.29236

©2017由美国肝病研究协会。

问题
肝病

第66卷，第6期，第1750-1765页，2017年12月
文章的对等评分为3

    1 INSERM U1052，法国里昂里昂癌症研究中心
    2里昂大学Claude-Bernard大学UMR_S1052，UCBL，法国里昂
    3普渡大学，西拉法叶，基础医学和普渡大学癌症研究中心
    4 CIRI国际传染病研究中心，EVIR，INSERM，U1111队，里昂大学Claude Bernard Lyon 1，法国里昂Univirsity of Lyon，法国里昂高等法院里昂高等师范学院UMR5308
    5法国里昂Civils de Lyon医院Croix-Rousse医院肝胃肠病科
    6 Labex DEVweCAN，法国里昂

    潜在的利益冲突：无需报告。
    † 这些作者同等贡献这项工作。

电子邮件：Adrien Foca（[email protected]），David Durantel（[email protected]）

*地址通讯和重申要求：
David Durantel博士
INSERM U1052
里昂癌症研究中心
里昂69008，法国
电子邮件：[email protected]
电话：+ 33-472-681-970 / + 33-472-681-959
要么
Ourania Andrisani，博士
基础医学部
普渡大学
West Lafayette，IN 47907 USA
电子邮件：[email protected]
电话：+ 1-765-494-8131

    这项工作得到了ANRS（法国国家艾滋病和病毒性肝炎研究机构，CSS4的几项赠款）和INSERM核心赠款给F.L.C.，F.Z.和D.D.的资助。这项工作也得到法国国家研究机构对F.Z.的管理，由里昂大学的DEVweCAN LABEX（ANR-10-LABX-0061）在Investissements d'Avenir（ANR-11-IDEX-0007）项目内予以支持。和D.D.和国立卫生研究院拨款（DK044533-19）给O.A.欧洲研究委员会（ERC-2008-AdG-233130-HEPCENT）和LabEx Ecofect（ANR-11-LABX-0048）进一步支持了FL Cosset实验室的工作。 A.D.获得法国驻华盛顿特区大使馆的Chateaubriand研究金。



慢性乙型肝炎病毒（HBV）感染是肝细胞癌（HCC）的主要危险因素，目前治疗慢性乙型肝炎和HCC的疗法并不理想。在此，我们鉴定了细胞丝氨酸/苏氨酸Polo-like-kinase 1（PLK1）作为HBV复制的正效应物。本研究的目的是证明PLK1在HBV生物合成中的前病毒作用，并验证PLK1抑制是一种潜在的抗病毒策略。为此，我们采用与原代人肝细胞（PHH）生理相关的HBV感染模型和分化的HepaRG细胞与药理学PLK1抑制剂，小干扰RNA（siRNA）介导的敲低和组成性活化的PLK1（PLK1CA）的过度表达。此外，还使用人源化的Fah - / - / Rag2 - / - / Il2rg - / - （FRG）小鼠模型来确定PLK1抑制剂BI-2536对HBV感染的体内抗病毒作用。最后，体外PLK1激酶测定和定点诱变用于证明HBV核心蛋白（HBc）是PLK1底物。我们证明HBV感染激活PHHs中的细胞PLK1和分化的HepaRG细胞。由BI-2536抑制PLK1或siRNA介导的敲低抑制HBV DNA生物合成，而PLK1CA的过度表达增加，表明PLK1对病毒生物合成的影响是特异性的，并且PLK1是前病毒细胞因子。重要的是，BI-2536给予HBV感染的人源化肝FRG小鼠强烈抑制HBV感染，验证PLK1作为体内抗病毒靶标。 PLK1的原病毒作用与核衣壳的生物发生相关，因为BI-2536导致其细胞内形成/积累减少。在这方面，我们的研究在体外鉴定了HBc为PLK1底物，并在该蛋白质上定位了PLK1磷酸化位点。结论：PLK1是一种前病毒宿主因子，可作为HBV感染和HBV介导癌变的联合抗病毒和抗肿瘤策略的靶点。 （Hepatology 2017; 66：1750-1765）