Genome-wide RNAi screen identifies human host factors crucial for influenza virus replication
Genome-wide RNAi screen identifies human host factors crucial for influenza virus replication
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DOI：
10.1038/nature08760
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发表时间：
2010-02-11
期刊：
影响因子：
64.8
通讯作者：
Meyer, Thomas F.
中科院分区：
文献类型：
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作者：
Karlas, Alexander;Machuy, Nikolaus;Meyer, Thomas F.
甲型流感病毒导致季节性流行病和反复发生的流行病，对全球公共卫生构成威胁(1)。高突变率有利于病毒逃逸突变体的产生，导致针对病毒编码目标的疫苗和药物可能无效(2)。相比之下，针对宿主细胞决定簇暂时对宿主来说是不必要的，但对病毒复制至关重要，可以防止病毒逃逸。在此，我们报告在全基因组 RNA 干扰 (RNAi) 筛选中发现了影响甲型流感病毒复制的 287 个人类宿主细胞基因。通过独立测定，我们确认了 168 个命中 (59%) 抑制了地方性 H1N1（119 个命中）或当前大流行的猪源甲型流感病毒株（121 个命中），重叠率为 60%。值得注意的是，这些常见命中的一部分对于高致病性禽 H5N1 病毒株的复制也至关重要。对几个因素的深入分析提供了对其感染阶段相关性的见解。值得注意的是，SON DNA 结合蛋白 (SON)(3) 被发现对于感染早期流感病毒颗粒正常运输到晚期内体非常重要。我们还表明，CDC 样激酶 1 (CLK1)(4) 的小分子抑制剂可将流感病毒复制减少两个数量级以上，这种效应与病毒 M2 信使 RNA 剪接受损有关。此外，感染流感病毒的p27(-/-)（细胞周期蛋白依赖性激酶抑制剂1B；Cdkn1b）小鼠在肺部积累的病毒滴度显着降低，为该基因的重要性提供了体内证据。因此，我们的结果强调了全基因组 RNAi 筛选对于剖析病毒-宿主相互作用和识别多种流感病毒药物靶点的效力。
Influenza A virus, being responsible for seasonal epidemics and reoccurring pandemics, represents a worldwide threat to public health(1). High mutation rates facilitate the generation of viral escape mutants, rendering vaccines and drugs directed against virus-encoded targets potentially ineffective(2). In contrast, targeting host cell determinants temporarily dispensable for the host but crucial for virus replication could prevent viral escape. Here we report the discovery of 287 human host cell genes influencing influenza A virus replication in a genome-wide RNA interference (RNAi) screen. Using an independent assay we confirmed 168 hits (59%) inhibiting either the endemic H1N1 (119 hits) or the current pandemic swine-origin (121 hits) influenza A virus strains, with an overlap of 60%. Notably, a subset of these common hits was also essential for replication of a highly pathogenic avian H5N1 strain. In-depth analyses of several factors provided insights into their infection stage relevance. Notably, SON DNA binding protein (SON)(3) was found to be important for normal trafficking of influenza virions to late endosomes early in infection. We also show that a small molecule inhibitor of CDC-like kinase 1 (CLK1)(4) reduces influenza virus replication by more than two orders of magnitude, an effect connected with impaired splicing of the viral M2 messenger RNA. Furthermore, influenza-virus-infected p27(-/-) (cyclin-dependent kinase inhibitor 1B; Cdkn1b) mice accumulated significantly lower viral titres in the lung, providing in vivo evidence for the importance of this gene. Thus, our results highlight the potency of genome-wide RNAi screening for the dissection of virus-host interactions and the identification of drug targets for a broad range of influenza viruses.