Cell ( IF 64.5 ) Pub Date : 2020-10-20 , DOI: 10.1016/j.cell.2020.10.028 Jin Wei 1 , Mia Madel Alfajaro 1 , Peter C DeWeirdt 2 , Ruth E Hanna 2 , William J Lu-Culligan 3 , Wesley L Cai 4 , Madison S Strine 1 , Shang-Min Zhang 4 , Vincent R Graziano 1 , Cameron O Schmitz 1 , Jennifer S Chen 1 , Madeleine C Mankowski 1 , Renata B Filler 1 , Neal G Ravindra 5 , Victor Gasque 5 , Fernando J de Miguel 6 , Ajinkya Patil 7 , Huacui Chen 4 , Kasopefoluwa Y Oguntuyo 8 , Laura Abriola 9 , Yulia V Surovtseva 9 , Robert C Orchard 10 , Benhur Lee 8 , Brett D Lindenbach 11 , Katerina Politi 12 , David van Dijk 5 , Cigall Kadoch 7 , Matthew D Simon 13 , Qin Yan 6 , John G Doench 2 , Craig B Wilen 14
Identification of host genes essential for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection may reveal novel therapeutic targets and inform our understanding of coronavirus disease 2019 (COVID-19) pathogenesis. Here we performed genome-wide CRISPR screens in Vero-E6 cells with SARS-CoV-2, Middle East respiratory syndrome CoV (MERS-CoV), bat CoV HKU5 expressing the SARS-CoV-1 spike, and vesicular stomatitis virus (VSV) expressing the SARS-CoV-2 spike. We identified known SARS-CoV-2 host factors, including the receptor ACE2 and protease Cathepsin L. We additionally discovered pro-viral genes and pathways, including HMGB1 and the SWI/SNF chromatin remodeling complex, that are SARS lineage and pan-coronavirus specific, respectively. We show that HMGB1 regulates ACE2 expression and is critical for entry of SARS-CoV-2, SARS-CoV-1, and NL63. We also show that small-molecule antagonists of identified gene products inhibited SARS-CoV-2 infection in monkey and human cells, demonstrating the conserved role of these genetic hits across species. This identifies potential therapeutic targets for SARS-CoV-2 and reveals SARS lineage-specific and pan-CoV host factors that regulate susceptibility to highly pathogenic CoVs.
中文翻译:
全基因组 CRISPR 筛选揭示了 SARS-CoV-2 感染的关键宿主因素
鉴定严重急性呼吸系统综合症冠状病毒 2 (SARS-CoV-2) 感染所必需的宿主基因可能会揭示新的治疗靶点,并有助于我们了解冠状病毒病 2019 (COVID-19) 的发病机制。在这里,我们使用 SARS-CoV-2、中东呼吸综合征冠状病毒 (MERS-CoV)、表达 SARS-CoV-1 刺突的蝙蝠冠状病毒 HKU5 和水疱性口炎病毒 (VSV) 在 Vero-E6 细胞中进行全基因组 CRISPR 筛选表达 SARS-CoV-2 尖峰。我们确定了已知的 SARS-CoV-2 宿主因子,包括受体 ACE2 和蛋白酶组织蛋白酶 L。我们还发现了促病毒基因和通路,包括HMGB1和 SWI/SNF 染色质重塑复合体,它们是 SARS 谱系和泛冠状病毒特有的, 分别。我们表明 HMGB1 调节ACE2表达,对 SARS-CoV-2、SARS-CoV-1 和 NL63 的进入至关重要。我们还表明,已识别基因产物的小分子拮抗剂抑制了 SARS-CoV-2 在猴子和人类细胞中的感染,证明了这些基因命中在物种间的保守作用。这确定了 SARS-CoV-2 的潜在治疗靶点,并揭示了 SARS 谱系特异性和泛 CoV 宿主因子,这些因子调节对高致病性 CoV 的易感性。