Zhang Qi, Chen Catherine Zhengzheng, Swaroop Manju, Xu Miao, Wang Lihui, Lee Juhyung, Wang Amy Qiu, Pradhan Manisha, Hagen Natalie, Chen Lu, Shen Min, Luo Zhiji, Xu Xin, Xu Yue, Huang Wenwei, Zheng Wei, Ye Yihong
Laboratory of Molecular Biology, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD, 20892, USA.
National Center for Advancing Translational Sciences, National Institutes of Health, Rockville, MD, 20850, USA.
Cell Discov. 2020 Nov 4;6(1):80. doi: 10.1038/s41421-020-00222-5.
The cell entry of SARS-CoV-2 has emerged as an attractive drug repurposing target for COVID-19. Here we combine genetics and chemical perturbation to demonstrate that ACE2-mediated entry of SARS-Cov and CoV-2 requires the cell surface heparan sulfate (HS) as an assisting cofactor: ablation of genes involved in HS biosynthesis or incubating cells with a HS mimetic both inhibit Spike-mediated viral entry. We show that heparin/HS binds to Spike directly, and facilitates the attachment of Spike-bearing viral particles to the cell surface to promote viral entry. We screened approved drugs and identified two classes of inhibitors that act via distinct mechanisms to target this entry pathway. Among the drugs characterized, Mitoxantrone is a potent HS inhibitor, while Sunitinib and BNTX disrupt the actin network to indirectly abrogate HS-assisted viral entry. We further show that drugs of the two classes can be combined to generate a synergized activity against SARS-CoV-2-induced cytopathic effect. Altogether, our study establishes HS as an attachment factor that assists SARS coronavirus cell entry and reveals drugs capable of targeting this important step in the viral life cycle.
严重急性呼吸综合征冠状病毒2（SARS-CoV-2）的细胞进入已成为治疗2019冠状病毒病（COVID-19）颇具吸引力的药物重新利用靶点。在此，我们结合遗传学和化学干扰方法，证明血管紧张素转换酶2（ACE2）介导的SARS-CoV和SARS-CoV-2进入需要细胞表面硫酸乙酰肝素（HS）作为辅助因子：敲除参与HS生物合成的基因或用HS模拟物处理细胞均可抑制刺突蛋白介导的病毒进入。我们发现肝素/HS直接与刺突蛋白结合，并促进携带刺突蛋白的病毒颗粒附着于细胞表面以促进病毒进入。我们筛选了已批准的药物，并确定了两类通过不同机制作用于该进入途径的抑制剂。在所研究的药物中，米托蒽醌是一种有效的HS抑制剂，而舒尼替尼和BNTX可破坏肌动蛋白网络以间接消除HS辅助的病毒进入。我们进一步表明，这两类药物可以联合使用，以产生针对SARS-CoV-2诱导的细胞病变效应的协同活性。总之，我们的研究确定HS为辅助SARS冠状病毒进入细胞的附着因子，并揭示了能够靶向病毒生命周期这一重要步骤的药物。
