Ng Teresa I, Mo Hongmei, Pilot-Matias Tami, He Yupeng, Koev Gennadiy, Krishnan Preethi, Mondal Rubina, Pithawalla Ron, He Wenping, Dekhtyar Tanya, Packer Jeremy, Schurdak Mark, Molla Akhteruzzaman
Global Pharmaceutical Research and Development, Antiviral Research, Abbott Laboratories, 200 Abbott Park Road, Abbott Park, IL 60064, USA.
Hepatology. 2007 Jun;45(6):1413-21. doi: 10.1002/hep.21608.
Hepatitis C virus (HCV) replication is highly dependent on host cell factors. Identification of these host factors not only facilitates understanding of the biology of HCV infection but also enables the discovery of novel targets for anti-HCV therapy. To identify host genes important for HCV RNA replication, we screened a library of small interfering RNA (siRNA) that targets approximately 4,000 human genes in Huh7-derived EN5-3 cells harboring an HCV subgenomic replicon with the nonstructural region NS3-NS5B from the 1b-N strain. Nine cellular genes that potentially regulate HCV replication were identified in this screen. Silencing of these genes resulted in inhibition of HCV replication by more than 60% and exhibited minimal toxicity. Knockdown of host gene expression by these siRNAs was confirmed at the RNA level and, in some instances, at the protein level. The level of siRNA silencing of these host genes correlated well with inhibition of HCV. These genes included those that encoded a G-protein coupled receptor (TBXA2R), a membrane protein (LTbeta), an adapter protein (TRAF2), 2 transcription factors (RelA and NFkappaB2), 2 protein kinases (MKK7 and SNARK), and 2 closely related transporter proteins (SLC12A4 and SLC12A5). Of interest, some of these genes are members of the tumor necrosis factor/lymphotoxin signaling pathway.
Findings of this study may provide important information for understanding HCV replication. In addition, these cellular genes may constitute a novel set of targets for HCV antiviral therapy.
丙型肝炎病毒（HCV）复制高度依赖宿主细胞因子。鉴定这些宿主因子不仅有助于理解HCV感染的生物学机制，还能发现抗HCV治疗的新靶点。为了鉴定对HCV RNA复制重要的宿主基因，我们在携带来自1b-N株的非结构区NS3-NS5B的HCV亚基因组复制子的Huh7衍生的EN5-3细胞中筛选了一个靶向约4000个人类基因的小干扰RNA（siRNA）文库。在该筛选中鉴定出九个可能调节HCV复制的细胞基因。这些基因的沉默导致HCV复制受到超过60%的抑制，且毒性极小。通过这些siRNA对宿主基因表达的敲低在RNA水平以及某些情况下在蛋白质水平得到证实。这些宿主基因的siRNA沉默水平与对HCV的抑制密切相关。这些基因包括编码G蛋白偶联受体（TBXA2R）、膜蛋白（LTβ）、衔接蛋白（TRAF2）、2种转录因子（RelA和NFκB2）、2种蛋白激酶（MKK7和SNARK）以及2种密切相关的转运蛋白（SLC12A4和SLC12A5）的基因。有趣的是，其中一些基因是肿瘤坏死因子/淋巴毒素信号通路的成员。
本研究结果可能为理解HCV复制提供重要信息。此外，这些细胞基因可能构成一组新的HCV抗病毒治疗靶点。
