Heparan sulfate assists SARS-CoV-2 in cell entry and can be targeted by approved drugs in vitro

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 © 2020, This is a U S Government work and not under copyright protection in the U S ;foreign copyright protection may apply