ACE2 is the critical in vivo receptor for SARS-CoV-2 in a novel COVID-19 mouse model with TNF- and IFNγ-driven immunopathology

  1. Riem Gawish
  2. Philipp Starkl
  3. Lisabeth Pimenov
  4. Anastasiya Hladik
  5. Karin Lakovits
  6. Felicitas Oberndorfer
  7. Shane JF Cronin
  8. Anna Ohradanova-Repic
  9. Gerald Wirnsberger
  10. Benedikt Agerer
  11. Lukas Endler
  12. Tümay Capraz
  13. Jan W Perthold
  14. Domagoj Cikes
  15. Rubina Koglgruber
  16. Astrid Hagelkruys
  17. Nuria Montserrat
  18. Ali Mirazimi
  19. Louis Boon
  20. Hannes Stockinger
  21. Andreas Bergthaler
  22. Chris Oostenbrink
  23. Josef M Penninger
  24. Sylvia Knapp

  • Curated by eLife

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    Evaluation Summary:

    To establish a mouse model for the SARS-CoV-2 infection, Gawish and colleagues performed serial passage of a human virus isolate in mice. They show that the mouse-adapted SARS-CoV-2 variant remains dependent on ACE2 for efficient infection and recapitulates some clinical characteristics of COVID-19. In addition, they demonstrate that inhalation of recombinant ACE2 protected mice from mouse COVID-19 suggesting this this model will be useful for the testing of antiviral agents.

    (This preprint has been reviewed by eLife. We include the public reviews from the reviewers here; the authors also receive private feedback with suggested changes to the manuscript. Reviewer #1 agreed to share their name with the authors.)

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Abstract

Despite tremendous progress in the understanding of COVID-19, mechanistic insight into immunological, disease-driving factors remains limited. We generated maVie16, a mouse-adapted SARS-CoV-2, by serial passaging of a human isolate. In silico modeling revealed how only three Spike mutations of maVie16 enhanced interaction with murine ACE2. maVie16 induced profound pathology in BALB/c and C57BL/6 mice, and the resulting mouse COVID-19 (mCOVID-19) replicated critical aspects of human disease, including early lymphopenia, pulmonary immune cell infiltration, pneumonia, and specific adaptive immunity. Inhibition of the proinflammatory cytokines IFNγ and TNF substantially reduced immunopathology. Importantly, genetic ACE2-deficiency completely prevented mCOVID-19 development. Finally, inhalation therapy with recombinant ACE2 fully protected mice from mCOVID-19, revealing a novel and efficient treatment. Thus, we here present maVie16 as a new tool to model COVID-19 for the discovery of new therapies and show that disease severity is determined by cytokine-driven immunopathology and critically dependent on ACE2 in vivo.

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  1. Version published to 10.7554/elife.74623 on eLife
  2. eLife

    Evaluation Summary:

    To establish a mouse model for the SARS-CoV-2 infection, Gawish and colleagues performed serial passage of a human virus isolate in mice. They show that the mouse-adapted SARS-CoV-2 variant remains dependent on ACE2 for efficient infection and recapitulates some clinical characteristics of COVID-19. In addition, they demonstrate that inhalation of recombinant ACE2 protected mice from mouse COVID-19 suggesting this this model will be useful for the testing of antiviral agents.

    (This preprint has been reviewed by eLife. We include the public reviews from the reviewers here; the authors also receive private feedback with suggested changes to the manuscript. Reviewer #1 agreed to share their name with the authors.)

  3. eLife

    Reviewer #1 (Public Review):

    Small animal models are very useful to study the pathogenesis of human diseases and for preclinical studies of potential therapeutic agents. In the present study, Gawish et al. passaged SARS-CoV-2 in mice to generate a mouse-adapted virus strain (maVie16) that replicates efficiently in mice and induces inflammation-driven pulmonary disease. They show that the mouse-adapted virus remains strictly dependent on ACE2 and demonstrate protective effects of inhibition of proinflammatory cytokines and inhaled recombinant ACE2. The authors conclude that they established a novel mouse model for COVID-19 that recapitulates many features of human disease and allows the testing of antiviral agents.

    As mentioned in the manuscript, several other groups have previously reported the development of mouse models for COVID-19. Initially, transgenic mice expressing the human ACE2 receptors were generated. However, these are extremely susceptible to SARS-CoV-2 infection and show a distinct pathology. Others have also generated mouse-adapted SARS-CoV-2 strains which share some features of the present maVie16 strain. Thus, not all aspects of the present study are entirely novel. Strength of the present study are that the authors show that, similarly to COVID-19 in humans, disease severity is to some extent determined by cytokine-driven immunopathology. Using ACE2 knockout animals, the authors also show that SARS-CoV-2 replication remains strictly dependent on ACE2. While this is as expected definitive experimental evidence is nonetheless important. Additional strengths are the evidence the pDCs play are key role in the response to infection and the demonstration that maVie16 strain in duces mCOVID-19 in both BALB/c and C57BL/6 mice. Altogether, the results suggest that this novel mouse-adapted SARS-CoV-2 strain will be useful for studies on viral pathogenesis and testing of antiviral agents.

  4. eLife

    Reviewer #2 (Public Review):

    A SARS-CoV-2 isolate was passaged in Balb/c mice and it is demonstrated that viral load and cytokine induction increased with passage number while body weight decreased. Further, mutations acquired during passaging are reported and evidence obtained in silico is provided that these mutations increased binding of the viral spike protein to murine ACE2. The virus is shown to cause lethal disease in Balb/c but not C57/BL6 mice, although the latter show body weight loss and transient pneumonia. The impact of infection on blood cell populations and cytokine release as well as adaptive immune responses is reported. In addition, elevated levels of IFNgamma and TNF were measured and antibodies targeting these factors are shown to protect from body weight loss. Finally, evidence is provided that ACE2 is required for viral spread in the lung and induction of disease and these processes were blocked by soluble ACE2. The findings are solid and the model will be helpful to the field.

  5. ScreenIT

    SciScore for 10.1101/2021.08.09.455606: (What is this?)

    Please note, not all rigor criteria are appropriate for all manuscripts.

    Table 1: Rigor

    Ethicsnot detected.
    Sex as a biological variablenot detected.
    Randomizationnot detected.
    Blindingnot detected.
    Power Analysisnot detected.

    Table 2: Resources

    No key resources detected.

    Results from OddPub: We did not detect open data. We also did not detect open code. Researchers are encouraged to share open data when possible (see Nature blog).

    Results from LimitationRecognizer: An explicit section about the limitations of the techniques employed in this study was not found. We encourage authors to address study limitations.

    Results from TrialIdentifier: No clinical trial numbers were referenced.

    Results from Barzooka: We did not find any issues relating to the usage of bar graphs.

    Results from JetFighter: We did not find any issues relating to colormaps.

    Results from rtransparent:
    • Thank you for including a conflict of interest statement. Authors are encouraged to include this statement when submitting to a journal.
    • Thank you for including a funding statement. Authors are encouraged to include this statement when submitting to a journal.
    • No protocol registration statement was detected.

    Results from scite Reference Check: We found no unreliable references.

    About SciScore

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  6. Version published to 10.1101/2021.08.09.455606v1 on bioRxiv