TMPRSS2 and TMPRSS4 promote SARS-CoV-2 infection of human small intestinal enterocytes

  1. Ruochen Zang
  2. Maria Florencia Gomez Castro
  3. Broc T. McCune
  4. Qiru Zeng
  5. Paul W. Rothlauf
  6. Naomi M. Sonnek
  7. Zhuoming Liu
  8. Kevin F. Brulois
  9. Xin Wang
  10. Harry B. Greenberg
  11. Michael S. Diamond
  12. Matthew A. Ciorba
  13. Sean P. J. Whelan
  14. Siyuan Ding

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Abstract

The SARS-CoV-2 virus infects cultured ACE2-expressing human enterocytes aided by the TMPRSS2 and TMPRSS4 serine proteases.

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  1. ScreenIT

    SciScore for 10.1101/2020.04.21.054015: (What is this?)

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

    Table 1: Rigor

    Institutional Review Board StatementConsent: All enteroids were derived from de-identified tissue from healthy, non-IBD subjects undergoing colonoscopy and provided informed consent at Stanford University and Washington University in St. Louis.
    Randomizationnot detected.
    Blindingnot detected.
    Power Analysisnot detected.
    Sex as a biological variablenot detected.

    Table 2: Resources

    Antibodies
    SentencesResources
    Samples were then stained with the following primary antibodies or fluorescent dyes: ACE2 (sc-390851 AF594, Santa Cruz), DAPI (P36962, Thermo Fisher), SARS-CoV-2 S (CR3022 human monoclonal antibody (51)), Villin (sc-58897 AF488, Santa Cruz), and phalloidin (Alexa 647-conjugated, Thermo Fisher)
    ACE2
    suggested: (Santa Cruz Biotechnology Cat# sc-390851, RRID:AB_2861379)
    P36962
    suggested: None
    Villin
    suggested: (Santa Cruz Biotechnology Cat# sc-58897, RRID:AB_2304475)
    Software and Algorithms
    SentencesResources
    Images were analyzed by Volocity v6.3 (PerkinElmer) and quantification was determined by CellProfiler (Broad Institute)
    Volocity
    suggested: (Volocity 3D Image Analysis Software, RRID:SCR_002668)
    Imputed expression values were calculated using a customized implementation (https://github.com/kbrulois/magicBatch) of the MAGIC (Markov Affinity-based Graph Imputation of Cells) algorithm (55) and optimized parameters (t = 2, k = 9, ka = 3).
    MAGIC
    suggested: (Magic, RRID:SCR_006406)
    Pooled expression of all viral genes was calculated using the AddModuleScore function from the Seurat package.
    Seurat
    suggested: (SEURAT, RRID:SCR_007322)

    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: We detected the following sentences addressing limitations in the study:
    Due to the limitation of sample size, we cannot definitely conclude that fecal-oral transmission of COVID-19 does not exist. However, our data is consistent with the previous reports from SARS-CoV and MERS-CoV publications which concluded that despite long duration of viral RNA shedding, no infectious virus could be recovered from the patients’ feces (41, 42). Intriguingly, the small intestinal fluids, which contain taurocholic acid sodium salt and lecithin, important surfactant components of the bile, did not inactivate virus as expected (Fig. 5A). In addition, we also observed that the virus titers did not drop in the presence of 0.03% bovine bile (data not shown), the highest concentration that did not induce cytotoxicity, contradicting the known fragility of lipid bilayers and sensitivity to bile salts which serve as detergents in gut. Recent studies showed a heavy glycosylation of SARS-CoV-2 S protein (43) and it is possible the sugar coating conferred some stability against the enzymatic digestion and bile salt solubilization. Two recent publications did demonstrate that SARS-CoV-2 has unique stability in the environment, at least in a laboratory setting (44, 45). Further work is needed to determine which bioactive(s) in the colonic fluids that are absent from small intestinal fluids function to potentially disrupt viral lipid membranes in vivo. Mechanistically, recent work from multiple groups showed a key role of TMPRSS2 in cleaving SARS-CoV-2 S protein. Here, we prov...

    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.

    About SciScore

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  2. Version published to 10.1126/sciimmunol.abc3582
  3. preLights

    Excerpt

    A gut feeling: intestinal organoids model COVID-19 pathogenesis illuminating a novel mode of infection where SARS-CoV-2 invades human enterocytes via TMPRSS2 and TMPRSS4 action

  4. Version published to 10.1101/2020.04.21.054015 on bioRxiv