Sars-cov-2 requires cholesterol for viral entry and pathological syncytia formation

David W. Sanders; Chanelle C. Jumper; Paul J. Ackerman; Dan Bracha; Anita Donlic; Hahn Kim; Devin Kenney; Ivan Castello-Serrano; Saori Suzuki; Tomokazu Tamura; Alexander H. Tavares; Mohsan Saeed; Alex S. Holehouse; Alexander Ploss; Ilya Levental; Florian Douam; Robert F. Padera; Bruce D. Levy; Clifford P. Brangwynne

doi:10.7554/ELIFE.65962

Sars-cov-2 requires cholesterol for viral entry and pathological syncytia formation

David W. Sanders, Chanelle C. Jumper, Paul J. Ackerman, Dan Bracha, Anita Donlic, Hahn Kim, Devin Kenney, Ivan Castello-Serrano, Saori Suzuki, Tomokazu Tamura, Alexander H. Tavares, Mohsan Saeed, Alex S. Holehouse, Alexander Ploss, Ilya Levental, Florian Douam, Robert F. Padera, Bruce D. Levy, Clifford P. Brangwynne

Research output: Contribution to journal › Article › peer-review

91 Scopus citations

Abstract

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 quantitative cell biology approaches, the screen reveals an essential role for biophysical aspects of the membrane, particularly cholesterol-rich regions, in spike-mediated fusion, which extends to replication-competent SARS-CoV-2 isolates. Our findings potentially 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.

Original language	English (US)
Article number	e65962
Journal	eLife
Volume	10
DOIs	https://doi.org/10.7554/ELIFE.65962
State	Published - Apr 2021

All Science Journal Classification (ASJC) codes

Neuroscience(all)
Biochemistry, Genetics and Molecular Biology(all)
Immunology and Microbiology(all)

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10.7554/ELIFE.65962

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Sanders, D. W., Jumper, C. C., Ackerman, P. J., Bracha, D., Donlic, A., Kim, H., Kenney, D., Castello-Serrano, I., Suzuki, S., Tamura, T., Tavares, A. H., Saeed, M., Holehouse, A. S., Ploss, A., Levental, I., Douam, F., Padera, R. F., Levy, B. D., & Brangwynne, C. P. (2021). Sars-cov-2 requires cholesterol for viral entry and pathological syncytia formation. eLife, 10, [e65962]. https://doi.org/10.7554/ELIFE.65962

@article{21d81a0174274e4f98a559f5228fc7d1,

title = "Sars-cov-2 requires cholesterol for viral entry and pathological syncytia formation",

abstract = "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 quantitative cell biology approaches, the screen reveals an essential role for biophysical aspects of the membrane, particularly cholesterol-rich regions, in spike-mediated fusion, which extends to replication-competent SARS-CoV-2 isolates. Our findings potentially 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.",

author = "Sanders, {David W.} and Jumper, {Chanelle C.} and Ackerman, {Paul J.} and Dan Bracha and Anita Donlic and Hahn Kim and Devin Kenney and Ivan Castello-Serrano and Saori Suzuki and Tomokazu Tamura and Tavares, {Alexander H.} and Mohsan Saeed and Holehouse, {Alex S.} and Alexander Ploss and Ilya Levental and Florian Douam and Padera, {Robert F.} and Levy, {Bruce D.} and Brangwynne, {Clifford P.}",

note = "Funding Information: We thank all Brangwynne Lab members for helpful discussion and critiques and Evangelos Gatzo-giannis for help with live cell microscopy. AD wishes to thank the Hargrove lab at Duke University, and particularly Sarah Wicks, for assistance and use of the ChemAxon analysis software, as well as Dr. Brittany Morgan for helpful discussions. This work was supported by Princeton COVID-19 research funds through the Office of the Dean for Research (CPB and AP labs); the Howard Hughes Medical Institute (CPB lab); a Boston University start-up fund and Peter Paul Career Development Professorship (FD); NIH (GM095467 and HL122531 to BDL; GM134949, GM124072, and GM120351 to IL); Volkswagen Foundation (IL); Human Frontiers Science Program (IL); a Burroughs Wellcome Fund Award for Investigators in Pathogenesis (AP); Longer Life Foundation—RGA/Washington University Collaboration (ASH); postdoctoral fellowship awards from the Uehara Memorial Foundation and JSPS Research Fellowships for Young Scientists (TT); from the SENSHIN Medical Research Foundation (S.S); and from the Natural Sciences and Engineering Research Council of Canada (CCJ). Publisher Copyright: {\textcopyright} Sanders et al.",

year = "2021",

month = apr,

doi = "10.7554/ELIFE.65962",

language = "English (US)",

volume = "10",

journal = "eLife",

issn = "2050-084X",

publisher = "eLife Sciences Publications",

}

Sanders, DW, Jumper, CC, Ackerman, PJ, Bracha, D, Donlic, A, Kim, H, Kenney, D, Castello-Serrano, I, Suzuki, S, Tamura, T, Tavares, AH, Saeed, M, Holehouse, AS, Ploss, A, Levental, I, Douam, F, Padera, RF, Levy, BD & Brangwynne, CP 2021, 'Sars-cov-2 requires cholesterol for viral entry and pathological syncytia formation', eLife, vol. 10, e65962. https://doi.org/10.7554/ELIFE.65962

TY - JOUR

T1 - Sars-cov-2 requires cholesterol for viral entry and pathological syncytia formation

AU - Sanders, David W.

AU - Jumper, Chanelle C.

AU - Ackerman, Paul J.

AU - Bracha, Dan

AU - Donlic, Anita

AU - Kim, Hahn

AU - Kenney, Devin

AU - Castello-Serrano, Ivan

AU - Suzuki, Saori

AU - Tamura, Tomokazu

AU - Tavares, Alexander H.

AU - Saeed, Mohsan

AU - Holehouse, Alex S.

AU - Ploss, Alexander

AU - Levental, Ilya

AU - Douam, Florian

AU - Padera, Robert F.

AU - Levy, Bruce D.

AU - Brangwynne, Clifford P.

N1 - Funding Information: We thank all Brangwynne Lab members for helpful discussion and critiques and Evangelos Gatzo-giannis for help with live cell microscopy. AD wishes to thank the Hargrove lab at Duke University, and particularly Sarah Wicks, for assistance and use of the ChemAxon analysis software, as well as Dr. Brittany Morgan for helpful discussions. This work was supported by Princeton COVID-19 research funds through the Office of the Dean for Research (CPB and AP labs); the Howard Hughes Medical Institute (CPB lab); a Boston University start-up fund and Peter Paul Career Development Professorship (FD); NIH (GM095467 and HL122531 to BDL; GM134949, GM124072, and GM120351 to IL); Volkswagen Foundation (IL); Human Frontiers Science Program (IL); a Burroughs Wellcome Fund Award for Investigators in Pathogenesis (AP); Longer Life Foundation—RGA/Washington University Collaboration (ASH); postdoctoral fellowship awards from the Uehara Memorial Foundation and JSPS Research Fellowships for Young Scientists (TT); from the SENSHIN Medical Research Foundation (S.S); and from the Natural Sciences and Engineering Research Council of Canada (CCJ). Publisher Copyright: © Sanders et al.

PY - 2021/4

Y1 - 2021/4

N2 - 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 quantitative cell biology approaches, the screen reveals an essential role for biophysical aspects of the membrane, particularly cholesterol-rich regions, in spike-mediated fusion, which extends to replication-competent SARS-CoV-2 isolates. Our findings potentially 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.

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 quantitative cell biology approaches, the screen reveals an essential role for biophysical aspects of the membrane, particularly cholesterol-rich regions, in spike-mediated fusion, which extends to replication-competent SARS-CoV-2 isolates. Our findings potentially 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.

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UR - http://www.scopus.com/inward/citedby.url?scp=85105793467&partnerID=8YFLogxK

U2 - 10.7554/ELIFE.65962

DO - 10.7554/ELIFE.65962

M3 - Article

C2 - 33890572

AN - SCOPUS:85105793467

SN - 2050-084X

VL - 10

JO - eLife

JF - eLife

M1 - e65962

ER -

Sars-cov-2 requires cholesterol for viral entry and pathological syncytia formation

Abstract

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