Cited by 815SARS-CoV-2 and COVID-19 ResearchCOVID-19 Clinical Research StudiesProtease and Inhibitor Mechanisms

TMPRSS2 and furin are both essential for proteolytic activation of SARS-CoV-2 in human airway cells

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Bestle, Dorothea, Heindl, Miriam Ruth, Limburg, Hannah, Van Lam van, Thuy, Pilgram, Oliver, Moulton, Hong, Stein, David A, Hardes, Kornelia +7 more·2020·Life Science Alliance

Abstract

The novel emerged SARS-CoV-2 has rapidly spread around the world causing acute infection of the respiratory tract (COVID-19) that can result in severe disease and lethality. For SARS-CoV-2 to enter cells, its surface glycoprotein spike (S) must be cleaved at two different sites by host cell proteases, which therefore represent potential drug targets. In the present study, we show that S can be cleaved by the proprotein convertase furin at the S1/S2 site and the transmembrane serine protease 2 (TMPRSS2) at the S2' site. We demonstrate that TMPRSS2 is essential for activation of SARS-CoV-2 S in Calu-3 human airway epithelial cells through antisense-mediated knockdown of TMPRSS2 expression. Furthermore, SARS-CoV-2 replication was also strongly inhibited by the synthetic furin inhibitor MI-1851 in human airway cells. In contrast, inhibition of endosomal cathepsins by E64d did not affect virus replication. Combining various TMPRSS2 inhibitors with furin inhibitor MI-1851 produced more potent antiviral activity against SARS-CoV-2 than an equimolar amount of any single serine protease inhibitor. Therefore, this approach has considerable therapeutic potential for treatment of COVID-19.

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  • The S protein is cleaved by the host cell protease furin into subunits S1 and S2, which support cellular entry.

    “The S protein is cleaved by the host cell protease furin into subunits (S1 and S2), which support cellular entry.66”

    From COVID-19 mRNA vaccines: Platforms and current developments (2022) · cites this paper as [66]

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Authors
Bestle, D., Heindl, M. R., Limburg, H., Van Lam van, T., Pilgram, O., Moulton, H., Stein, D. A., Hardes, K., Eickmann, M., Dolnik, O., Rohde, C., Klenk, H.-D., Garten, W., Steinmetzer, T., & Böttcher-Friebertshäuser, E.
Affiliations
  • Philipps University of Marburg (DE)
  • Institute of Pharmaceutical Chemistry, Philipps-University, Marburg, Germany
  • Oregon State University (US)
  • Institute of Virology, Philipps-University, Marburg, Germany
Venue
Life Science Alliance
Published
2020-07-23
Volume
3
Issue
9
Pages
e202000786
ISSN
2575-1077 (ISSN-L)
OpenAlex
W3044321933
Cited by
815
References
68
Citations by year
2026: 38 · 2025: 59 · 2024: 98 · 2023: 116 · 2022: 186 · 2021: 246 · 2020: 72
FWCI
20.39
Type
journal article
Language
en
Open access
Locations
  • Life Science Alliance (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
  • Fraunhofer-Publica (Fraunhofer-Gesellschaft) (submittedVersion)
Funders
  • LOEWE Center DRUID (D1)
  • Deutsche Forschungsgemeinschaft (SFB 1021, B07)
  • German Center for Infection Research
Topics
  • Health Sciences › Medicine › Infectious Diseases › SARS-CoV-2 and COVID-19 Research
  • Health Sciences › Medicine › Infectious Diseases › COVID-19 Clinical Research Studies
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cancer Research › Protease and Inhibitor Mechanisms
Keywords
SARS-CoV-2, COVID-19 pandemic, TMPRSS2, furin, spike protein cleavage, host cell proteases, cathepsin inhibition, E64d, antiviral drug targets, viral replication
MeSH terms
Betacoronavirus, SARS-CoV-2, Animals, Binding Sites, Cell Line, Chlorocebus aethiops, Humans, Serine Endopeptidases, Vero Cells, Virus Replication, Furin, Virus Internalization, Alveolar Epithelial Cells, HEK293 Cells, Proteolysis, Spike Glycoprotein, Coronavirus

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