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Polo-like-kinase 1 is a proviral host factor for hepatitis B virus replication
Hepatology ( IF 13.5 ) Pub Date : 2017-11-06 , DOI: 10.1002/hep.29236
Ahmed Diab 1, 2, 3 , Adrien Foca 1, 2 , Floriane Fusil 2, 4 , Thomas Lahlali 1, 2 , Pascal Jalaguier 1, 2 , Fouzia Amirache 4 , Lia N'Guyen 1, 2 , Nathalie Isorce 1, 2 , François-Loïc Cosset 2, 4 , Fabien Zoulim 1, 2, 5, 6 , Ourania Andrisani 3 , David Durantel 1, 2, 6
Affiliation  

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)

中文翻译:

Polo样激酶1是乙型肝炎病毒复制的前病毒宿主因子

慢性乙型肝炎病毒(HBV)感染是肝细胞癌(HCC)的主要危险因素,目前对慢性乙型肝炎和HCC的治疗并不理想。在这里,我们确定细胞丝氨酸/苏氨酸Polo样激酶1(PLK1)为HBV复制的积极效应。这项研究的目的是证明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,我们将原发性人类肝细胞(PHHs)和分化的HepaRG细胞与生理学相关的HBV感染模型与药理性PLK1抑制剂,小干扰RNA(siRNA)介导的敲除以及组成型活性PLK1(PLK1 CA)的过表达相结合。此外,人源化肝脏Fah -/- / Rag2 -/-/ Il2rg -/-(FRG)小鼠模型用于确定PLK1抑制剂BI-2536对体内HBV感染的抗病毒作用。最后,采用体外PLK1激酶测定和定点诱变来证明HBV核心蛋白(HBc)是PLK1底物。我们证明了HBV感染激活PHHs和分化的HepaRG细胞中的细胞PLK1。BI-2536或siRNA介导的敲低PLK1的抑制作用抑制了HBV DNA的生物合成,而PLK1 CA的过表达增加它,表明PLK1对病毒生物合成的作用是特异性的,并且PLK1是前病毒细胞因子。重要的是,BI-2536对HBV感染的人源化肝FRG小鼠的给药强烈抑制了HBV感染,从而验证了PLK1是体内的抗病毒靶标。PLK1的前病毒作用与核衣壳的生物发生有关,因为BI-2536导致其减少的细胞内形成/积累。在这方面,我们的研究在体外将HBc鉴定为PLK1底物并在该蛋白上绘制了PLK1磷酸化位点。结论: PLK1是一种前病毒宿主因子,可以作为针对HBV感染和HBV介导的致癌作用的联合抗病毒和抗肿瘤策略的靶标。(H流行病学; 2017年; 66:1750–1765)
更新日期:2017-11-21
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