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Elucidation of host factors and the associated pathways responsible for cellular permissiveness to hepatitis E virus replication and identification of the potential inhibitors

阐明导致细胞允许戊型肝炎病毒复制的宿主因素和相关途径以及潜在抑制剂的鉴定

基本信息

批准号：
22K15478
负责人：
Putu・Prathiwi・Primadharsini
金额：
$ 2.91万
依托单位：
Jichi Medical University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Early-Career Scientists
财政年份：
2022
资助国家：
日本
起止时间：
2022-04-01 至 2024-03-31
项目状态：
已结题

项目摘要

Hepatitis E virus (HEV) is increasingly recognized as the leading cause of acute hepatitis. In immunocompromised patients, HEV can cause chronic hepatitis. Currently, there is no specific anti-HEV drug available. Targeting cellular factors associated with HEV replication can be one of the methods to develop specific anti-HEV drug. In this context, cell culture is required. Subclones of PLC/PRF/5 cells have variable permissiveness to HEV replication even when inoculated with the same virus, suggesting that aside from viral factor itself, cellular factors might be involved in determining host susceptibility to HEV replication, and therefore, can be the target for development of specific anti-HEV drug.Subclones of a single PLC/PRF/5 cell line demonstrated up to 10,000-folds difference in the permissiveness to HEV replication. Based on the results of RNA microarray analysis of highly permissive and poorly permissive PLC/PRF/5 subclones, 15 upregulated genes and 15 downregulated genes were selected. Small interfering RNA (siRNA)-mediated gene silencing was performed on the upregulated and downregulated genes, followed by screening using eHEV-nanoKAZ (Primadharsini et al., J Virol 2022) and evaluation in cell culture. Silencing of any of four of upregulated genes in highly permissive subclone resulted in decreased HEV replication efficiency, while silencing of any of four downregulated genes in the poorly permissive subclone resulted in slightly increased HEV replication efficiency.

戊型肝炎病毒（HEV）越来越被认为是急性肝炎的主要原因。在免疫功能低下的患者中，HEV可引起慢性肝炎。目前，尚无特异性抗HEV药物可用。靶向与HEV复制相关的细胞因子可能是开发特异性抗HEV药物的方法之一。在这种情况下，需要细胞培养。PLC/PRF/5细胞亚克隆对HEV复制的容许性不同，表明除了病毒本身因素外，细胞因素可能参与决定宿主对HEV复制的易感性，因此可以作为开发特异性抗HEV药物的靶点。对HEV复制的容许性相差000倍。根据高允许和低允许PLC/PRF/5亚克隆的RNA微阵列分析结果，选择15个上调基因和15个下调基因。对上调和下调的基因进行小干扰RNA（siRNA）介导的基因沉默，然后使用eHEV-nanoKAZ进行筛选（Primadharsini等人，J Virol 2022）和在细胞培养中的评价。在高允许性亚克隆中，四个上调基因中的任何一个的沉默导致HEV复制效率降低，而在低允许性亚克隆中，四个下调基因中的任何一个的沉默导致HEV复制效率略微增加。