A functional genomic screen reveals novel host genes that mediate interferon-alpha's effects against hepatitis C virus.
A functional genomic screen reveals novel host genes that mediate interferon-alpha's effects against hepatitis C virus.
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DOI：
10.1016/j.jhep.2011.07.026
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发表时间：
2012-02
影响因子：
25.7
通讯作者：
Chung, Raymond T.
中科院分区：
文献类型：
--
作者：
Zhao, Hong;Lin, Wenyu;Kumthip, Kattareeya;Cheng, Du;Fusco, Dahlene N.;Hofmann, Oliver;Jilg, Nikolaus;Tai, Andrew W.;Goto, Kaku;Zhang, Leiliang;Hide, Winston;Jang, Jae Young;Peng, Lee F.;Chung, Raymond T.
IFN对HCV发挥抗病毒作用的确切机制尚未阐明。我们试图通过进行全基因组siRNA文库筛选来鉴定介导IFN-α抗病毒作用的宿主基因。采用HCV基因型1b复制子pRep-Feo进行高通量筛选。复制稳健性Z评分≥2.0的池在全长OR6复制子细胞中进入二次验证。然后用感染JFH1的Huh7.5.1细胞验证IFN-α处理下选定基因对HCV复制的拯救作用。我们使用全基因组siRNA文库鉴定并确认了93个参与IFN-α抗hcv效应的人类基因。基因本体分析显示，mRNA加工（23个基因，P=2.756e-22）、翻译起始（9个基因，P=2.42e-6）和IFN信号传导（5个基因，P=1.00e-3）是最富集的功能群。9个基因为U4/U6的组成部分。U5 tri-snRNP。我们证实，沉默T细胞识别的鳞状细胞癌抗原（SART1）， tri-snRNP的一个特异性因子，可以在复制子细胞和JFH1感染细胞中消除IFN-α对HCV的抑制作用。我们进一步发现SART1不是IFN-α诱导的，在JFH1感染模型中，SART1的抗hcv效应是通过有或没有IFN-α的干扰素刺激基因（ISGs）来调节的。我们通过全基因组siRNA筛选鉴定了93个介导IFN-α抗hcv作用的基因；分别有23个和9个基因参与mRNA加工和翻译起始。这些发现揭示了mRNA加工在抗病毒状态产生中的意想不到的作用，并为HCV治疗开发提供了新的途径。
The precise mechanisms by which IFN exerts its antiviral effect against HCV have not yet been elucidated. We sought to identify host genes that mediate the antiviral effect of IFN-α by conducting a whole-genome siRNA library screen. High throughput screening was performed using an HCV genotype 1b replicon, pRep-Feo. Those pools with replicate robust Z scores ≥ 2.0 entered secondary validation in full-length OR6 replicon cells. Huh7.5.1 cells infected with JFH1 were then used to validate the rescue efficacy of selected genes for HCV replication under IFN-α treatment. We identified and confirmed 93 human genes involved in the IFN-α anti-HCV effect using a whole-genome siRNA library. Gene ontology analysis revealed that mRNA processing (23 genes, P=2.756e-22), translation initiation (9 genes, P=2.42e-6), and IFN signaling (5 genes, P=1.00e-3) were the most enriched functional groups. Nine genes were components of U4/U6.U5 tri-snRNP. We confirmed that silencing squamous cell carcinoma antigen recognized by T cells (SART1), a specific factor of tri-snRNP, abrogates IFN-α's suppressive effects against HCV in both replicon cells and JFH1 infectious cells. We further found that SART1 was not an IFN-α inducible, and its anti-HCV effector in the JFH1 infectious model was through regulation of interferon stimulated genes (ISGs) with or without IFN-α. We identified 93 genes that mediate the anti-HCV effect of IFN-α through genome-wide siRNA screening; 23 and 9 genes were involved in mRNA processing and translation initiation, respectively. These findings reveal an unexpected role for mRNA processing in generation of the antiviral state, and suggest a new avenue for therapeutic development in HCV.
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影响因子：
5.4
作者：
Kohaar, Indu;Ploss, Alexander;Prokunina-Olsson, Ludmila
通讯作者：
Prokunina-Olsson, Ludmila
影响因子：
2.3
作者：
Kaur, S;Uddin, S;Platanias, LC
通讯作者：
Platanias, LC
影响因子：
14.9
作者：
Huang DW;Sherman BT;Tan Q;Kir J;Liu D;Bryant D;Guo Y;Stephens R;Baseler MW;Lane HC;Lempicki RA
通讯作者：
Lempicki RA
影响因子：
4.5
作者：
Liu, Sunbin;Rauhut, Reinhard;Luehrmann, Reinhard
通讯作者：
Luehrmann, Reinhard
影响因子：
4.4
作者：
Lin, Ren-Jye;Yu, Han-Pang;Lin, Yi-Ling
通讯作者：
Lin, Yi-Ling