PT  - JOURNAL ARTICLE
AU  - David W. Sanders
AU  - Chanelle C. Jumper
AU  - Paul J. Ackerman
AU  - Dan Bracha
AU  - Anita Donlic
AU  - Hahn Kim
AU  - Devin Kenney
AU  - Ivan Castello-Serrano
AU  - Saori Suzuki
AU  - Tomokazu Tamura
AU  - Alexander H. Tavares
AU  - Mohsan Saeed
AU  - Alex S. Holehouse
AU  - Alexander Ploss
AU  - Ilya Levental
AU  - Florian Douam
AU  - Robert F. Padera
AU  - Bruce D. Levy
AU  - Clifford P. Brangwynne
TI  - SARS-CoV-2 Requires Cholesterol for Viral Entry and Pathological Syncytia Formation
AID  - 10.1101/2020.12.14.422737
DP  - 2020 Jan 01
TA  - bioRxiv
PG  - 2020.12.14.422737
4099  - http://biorxiv.org/content/early/2020/12/14/2020.12.14.422737.short
4100  - http://biorxiv.org/content/early/2020/12/14/2020.12.14.422737.full
AB  - Many enveloped viruses induce multinucleated cells (syncytia), reflective of membrane fusion events caused by the same machinery that underlies viral entry. These syncytia are thought to facilitate replication and evasion of the host immune response. Here, we report that co-culture of human cells expressing the receptor ACE2 with cells expressing SARS-CoV-2 spike, results in synapse-like intercellular contacts that initiate cell-cell fusion, producing syncytia resembling those we identify in lungs of COVID-19 patients. To assess the mechanism of spike/ACE2-driven membrane fusion, we developed a microscopy-based, cell-cell fusion assay to screen ∼6000 drugs and >30 spike variants. Together with cell biological and biophysical approaches, the screen reveals an essential role for membrane cholesterol in spike-mediated fusion, which extends to replication-competent SARS-CoV-2 isolates. Our findings provide a molecular basis for positive outcomes reported in COVID-19 patients taking statins, and suggest new strategies for therapeutics targeting the membrane of SARS-CoV-2 and other fusogenic viruses.Cell-cell fusion at ACE2-spike clusters cause pathological syncytia in COVID-19Drug screen reveals critical role for membrane lipid composition in fusionSpike’s unusual membrane-proximal cysteines and aromatics are essential for fusionCholesterol tunes relative infectivity of SARS-CoV-2 viral particlesCompeting Interest StatementC.P.B. is a scientific founder and consultant for Nereid Therapeutics. A.S.H. is a consultant for Dewpoint Therapeutics.