SARS-CoV-2 requires cholesterol for viral entry and pathological syncytia formation

  1. David W Sanders
  2. Chanelle C Jumper
  3. Paul J Ackerman
  4. Dan Bracha
  5. Anita Donlic
  6. Hahn Kim
  7. Devin Kenney
  8. Ivan Castello-Serrano
  9. Saori Suzuki
  10. Tomokazu Tamura
  11. Alexander H Tavares
  12. Mohsan Saeed
  13. Alex S Holehouse
  14. Alexander Ploss
  15. Ilya Levental
  16. Florian Douam
  17. Robert F Padera
  18. Bruce D Levy
  19. Clifford P Brangwynne  Is a corresponding author
  1. Princeton University, United States
  2. Boston University, United States
  3. University of Virginia, United States
  4. Washington University School of Medicine, United States
  5. Harvard Medical School, United States

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.

Data availability

All data generated or analyzed during this study are included in the manuscript and supporting files with the exception of raw imaging data (>400,000 Nikon ND2 files), which is not feasible to post online given its massive size (>1.5 TB). This data is available from the lead contact upon request, assuming the interested party provides a server with sufficient storage capacity. Raw data (computed fusion scores) from the drug repurposing screen is available in Supplemental File 1; bioinformatics, Supplemental File 3.

Article and author information

Author details

  1. David W Sanders

    Princeton University, Princeton, United States
    Competing interests
    No competing interests declared.
  2. Chanelle C Jumper

    Princeton University, Princeton, United States
    Competing interests
    No competing interests declared.
  3. Paul J Ackerman

    Princeton University, Princeton, United States
    Competing interests
    No competing interests declared.
  4. Dan Bracha

    Princeton University, Princeton, United States
    Competing interests
    No competing interests declared.
  5. Anita Donlic

    Princeton University, Princeton, United States
    Competing interests
    No competing interests declared.
  6. Hahn Kim

    Princeton University, Princeton, United States
    Competing interests
    No competing interests declared.
  7. Devin Kenney

    Boston University, Boston, United States
    Competing interests
    No competing interests declared.
  8. Ivan Castello-Serrano

    University of Virginia, Charlottesville, United States
    Competing interests
    No competing interests declared.
  9. Saori Suzuki

    Princeton University, Princeton, United States
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-5233-6604
  10. Tomokazu Tamura

    Princeton University, Princeton, United States
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-1395-6610
  11. Alexander H Tavares

    Boston University, Boston, United States
    Competing interests
    No competing interests declared.
  12. Mohsan Saeed

    Boston University, Boston, United States
    Competing interests
    No competing interests declared.
  13. Alex S Holehouse

    Department of Biochemistry and Molecular Biophysics, Washington University School of Medicine, St Louis, United States
    Competing interests
    Alex S Holehouse, A.S.H. is a consultant for Dewpoint Therapeutics..
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-4155-5729
  14. Alexander Ploss

    Princeton University, Princeton, United States
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-9322-7252
  15. Ilya Levental

    University of Virginia, Charlottesville, United States
    Competing interests
    No competing interests declared.
  16. Florian Douam

    Princeton University, Princeton, United States
    Competing interests
    No competing interests declared.
  17. Robert F Padera

    Harvard Medical School, Boston, United States
    Competing interests
    No competing interests declared.
  18. Bruce D Levy

    Harvard Medical School, Boston, United States
    Competing interests
    No competing interests declared.
  19. Clifford P Brangwynne

    Princeton University, Princeton, United States
    For correspondence
    cbrangwy@princeton.edu
    Competing interests
    Clifford P Brangwynne, C.P.B. is a scientific founder and consultant for Nereid Therapeutics..
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-1350-9960

Funding

National Institute of General Medical Sciences (GM095467)

  • Bruce D Levy

National Heart, Lung, and Blood Institute (HL122531)

  • Bruce D Levy

National Institute of General Medical Sciences (GM134949)

  • Ilya Levental

National Institute of General Medical Sciences (GM124072)

  • Ilya Levental

Howard Hughes Medical Institute

  • Clifford P Brangwynne

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Ethics

Human subjects: Human pathology studies were performed with the approval of the Institutional Review Board at Brigham and Women's Hospital. Clinical autopsies with full anatomic dissection were performed on SARS-CoV-2 decedents by a board-certified anatomic pathologist (RFP) with appropriateinfectious precautions.

Reviewing Editor

  1. William A Prinz, National Institutes of Health, United States

Publication history

  1. Received: December 21, 2020
  2. Accepted: April 1, 2021
  3. Accepted Manuscript published: April 23, 2021 (version 1)
  4. Version of Record published: May 7, 2021 (version 2)

Copyright

© 2021, Sanders et al.

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

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  1. David W Sanders
  2. Chanelle C Jumper
  3. Paul J Ackerman
  4. Dan Bracha
  5. Anita Donlic
  6. Hahn Kim
  7. Devin Kenney
  8. Ivan Castello-Serrano
  9. Saori Suzuki
  10. Tomokazu Tamura
  11. Alexander H Tavares
  12. Mohsan Saeed
  13. Alex S Holehouse
  14. Alexander Ploss
  15. Ilya Levental
  16. Florian Douam
  17. Robert F Padera
  18. Bruce D Levy
  19. Clifford P Brangwynne
(2021)
SARS-CoV-2 requires cholesterol for viral entry and pathological syncytia formation
eLife 10:e65962.
https://doi.org/10.7554/eLife.65962
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