Department of Laboratory Medicine, Yale School of Medicine, New Haven, CT 06520, USA; Department of Immunobiology, Yale School of Medicine, New Haven, CT 06520, USA.
Genetic Perturbation Platform, Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA.
Cell. 2021 Jan 7;184(1):76-91.e13. doi: 10.1016/j.cell.2020.10.028. Epub 2020 Oct 20.
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.
鉴定严重急性呼吸综合征冠状病毒 2（SARS-CoV-2）感染所必需的宿主基因，可能揭示新的治疗靶点，并有助于我们了解 2019 年冠状病毒病（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 刺突蛋白的蝙蝠 CoV HKU5 和表达 SARS-CoV-2 刺突蛋白的水疱性口炎病毒（VSV）在 Vero-E6 细胞中进行了全基因组 CRISPR 筛选。我们鉴定出了已知的 SARS-CoV-2 宿主因子，包括受体 ACE2 和蛋白酶 Cathepsin L。我们还发现了促病毒基因和途径，包括 HMGB1 和 SWI/SNF 染色质重塑复合物，它们分别是 SARS 谱系和泛冠状病毒特异性的。我们表明，HMGB1 调节 ACE2 的表达，对于 SARS-CoV-2、SARS-CoV-1 和 NL63 的进入是至关重要的。我们还表明，鉴定出的基因产物的小分子拮抗剂抑制了猴和人细胞中的 SARS-CoV-2 感染，这表明这些遗传命中在物种间具有保守作用。这确定了 SARS-CoV-2 的潜在治疗靶点，并揭示了 SARS 谱系特异性和泛冠状病毒宿主因子，它们调节对高致病性 CoV 的易感性。
