Novel perspectives for hepatitis A virus therapy revealed by comparative analysis of hepatitis C virus and hepatitis A virus RNA replication
Novel perspectives for hepatitis A virus therapy revealed by comparative analysis of hepatitis C virus and hepatitis A virus RNA replication
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DOI：
10.1002/hep.27847
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发表时间：
2015-08-01
期刊：
影响因子：
13.5
通讯作者：
Lohmann, Volker
中科院分区：
文献类型：
--
作者：
Esser-Nobis, Katharina;Harak, Christian;Lohmann, Volker
甲型肝炎病毒（HAV）和丙型肝炎病毒（HCV）是两种正链RNA病毒，具有相似的生物学特性，但导致相反的感染结果，HAV总是被清除，而HCV在大多数感染中持续存在。为了更深入地了解复制，持久性和治疗的决定因素，我们建立了一个同质的细胞培养模型，允许彻底比较两种病毒的RNA复制。通过用HAV的亚基因组报告复制子以及不同HCV基因型的亚基因组报告复制子筛选不同的人肝来源的细胞系，我们发现Huh 7-Lunet细胞支持HAV和HCV-RNA复制，具有相似的效率，并且两种复制酶之间的干扰有限。HAV和HCV复制子对干扰素（IFN）的敏感性相似，但在细胞培养中建立持久复制的能力不同。与HCV相反，HAV独立于microRNA-122和磷脂酰肌醇4-激酶III和（PI 4KIII）复制。这两种病毒有效地抑制环孢菌素A和NIM 811，其非免疫抑制类似物，表明重叠依赖亲环素复制。然而，一组更广泛的抑制剂的分析表明，与丙型肝炎病毒，甲型肝炎病毒不依赖于亲环素A，而是对腺苷三磷酸结合盒转运蛋白和FK 506结合蛋白。最后，水飞蓟宾，但不是其修改的静脉制剂，有效地抑制HAV基因组复制在体外，表明口服水飞蓟宾作为一个潜在的治疗选择HAV感染。结论：我们建立了一个细胞培养模型，能够比较研究RNA复制的HAV和HCV在一个同质的细胞背景具有可比的复制效率。因此，我们确定了新的宿主细胞靶点和HAV的潜在治疗方案，并为未来的研究奠定了基础，以解开清除和持久性的决定因素。（肝病学2015;62：397-408
Hepatitis A virus (HAV) and hepatitis C virus (HCV) are two positive-strand RNA viruses sharing a similar biology, but causing opposing infection outcomes, with HAV always being cleared and HCV establishing persistence in the majority of infections. To gain deeper insight into determinants of replication, persistence, and treatment, we established a homogenous cell-culture model allowing a thorough comparison of RNA replication of both viruses. By screening different human liver-derived cell lines with subgenomic reporter replicons of HAV as well as of different HCV genotypes, we found that Huh7-Lunet cells supported HAV- and HCV-RNA replication with similar efficiency and limited interference between both replicases. HAV and HCV replicons were similarly sensitive to interferon (IFN), but differed in their ability to establish persistent replication in cell culture. In contrast to HCV, HAV replicated independently from microRNA-122 and phosphatidylinositol 4-kinase III and (PI4KIII). Both viruses were efficiently inhibited by cyclosporin A and NIM811, a nonimmunosuppressive analog thereof, suggesting an overlapping dependency on cyclophilins for replication. However, analysis of a broader set of inhibitors revealed that, in contrast to HCV, HAV does not depend on cyclophilin A, but rather on adenosine-triphosphate-binding cassette transporters and FK506-binding proteins. Finally, silibinin, but not its modified intravenous formulation, efficiently inhibited HAV genome replication in vitro, suggesting oral silibinin as a potential therapeutic option for HAV infections. Conclusion: We established a cell-culture model enabling comparative studies on RNA replication of HAV and HCV in a homogenous cellular background with comparable replication efficiency. We thereby identified new host cell targets and potential treatment options for HAV and set the ground for future studies to unravel determinants of clearance and persistence. (Hepatology 2015;62:397-408