SARS-CoV-2 Infection Depends on Cellular Heparan Sulfate and ACE2

Clausen, Thomas Mandel; Sandoval, Daniel R.; Spliid, Charlotte B.; Pihl, Jessica; Painter, Chelsea D.; Thacker, Bryan E.; Glass, Charles A.; Narayanan, Anoop; Majowicz, Sydney A.; Zhang, Yang; Torres, Jonathan L.; Golden, Gregory J.; Porell, Ryan; Garretson, Aaron F.; Laubach, Logan; Feldman, Jared; Yin, Xin; Pu, Yuan; Hauser, Blake; Caradonna, Timothy M.; Kellman, Benjamin P.; Martino, Cameron; Gordts, Philip L.S.M.; Leibel, Sandra L.; Chanda, Summit K.; Schmidt, Aaron G.; Godula, Kamil; Jose, Joyce; Corbett, Kevin D.; Ward, Andrew B.; Carlin, Aaron F.; Esko, Jeffrey D.

We show that SARS-CoV-2 spike protein interacts with cell surface heparan sulfate and angiotensin converting enzyme 2 (ACE2) through its Receptor Binding Domain. Docking studies suggest a putative heparin/heparan sulfate-binding site adjacent to the domain that binds to ACE2. In vitro, binding of ACE2 and heparin to spike protein ectodomains occurs independently and a ternary complex can be generated using heparin as a template. Contrary to studies with purified components, spike protein binding to heparan sulfate and ACE2 on cells occurs codependently. Unfractionated heparin, non-anticoagulant heparin, treatment with heparin lyases, and purified lung heparan sulfate potently block spike protein binding and infection by spike protein-pseudotyped virus and SARS-CoV-2 virus. These findings support a model for SARS-CoV-2 infection in which viral attachment and infection involves formation of a complex between heparan sulfate and ACE2. Manipulation of heparan sulfate or inhibition of viral adhesion by exogenous heparin may represent new therapeutic opportunities.