Blockade of TMPRSS2-mediated priming of SARS-CoV-2 by lactoferricin

  1. Anna Ohradanova-Repic
  2. Rostislav Skrabana
  3. Laura Gebetsberger
  4. Gabor Tajti
  5. Peter Baráth
  6. Gabriela Ondrovičová
  7. Romana Praženicová
  8. Nikola Jantova
  9. Patricia Hrasnova
  10. Hannes Stockinger
  11. Vladimir Leksa

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Abstract

In addition to vaccines, there is an urgent need for supplemental antiviral therapeutics to dampen the persistent COVID-19 pandemic caused by the severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2). The transmembrane protease serine 2 (TMPRSS2), that is responsible for proteolytic priming of the SARS-CoV-2 spike protein, appears as a rational therapeutic target. Accordingly, selective inhibitors of TMPRSS2 represent potential tools for prevention and treatment of COVID-19. Previously, we identified the human milk glycoprotein lactoferrin as a natural inhibitor of plasminogen conversion to plasmin, a serine protease homologous to TMPRSS2. Here, we tested whether lactoferrin and lactoferricin, a biologically active natural peptide produced by pepsin-mediated digestion of lactoferrin, together with synthetic peptides derived from lactoferrin, were able to block TMPRSS2 and SARS-CoV-2 infection. Particularly, we revealed that both lactoferricin and the N-terminal synthetic peptide pLF1 significantly inhibited: i) proteolytic activity of TMPRSS2 and plasmin, ii) proteolytic processing of the SARS-CoV-2 spike protein, and iii) SARS-CoV-2 infection of SARS-CoV-2-permissive cells. Thus, natural and synthetic peptides derived from lactoferrin represent feasible candidates for supporting prevention and treatment of COVID-19.

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  1. Version published to 10.3389/fimmu.2022.958581
  2. ScreenIT

    SciScore for 10.1101/2021.12.20.473447: (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.
    Cell Line Authenticationnot detected.

    Table 2: Resources

    Antibodies
    SentencesResources
    The reaction mixtures were analyzed by immunoblotting with a specific anti-S-protein monoclonal antibody (mAb) (NovusBio, Littleton, CO).
    anti-S-protein
    suggested: None
    Experimental Models: Cell Lines
    SentencesResources
    SARS-CoV-2 infection and In-Cell ELISA: The African green monkey kidney-derived Vero cells (ATCC CCL-81TM) were seeded into 96-well flat-bottom plates (1×104 cells, 80 μl/well) in Dulbecco’s Modified Eagle’s medium (DMEM), high glucose, with GlutaMAX and sodium pyruvate (Gibco/Thermo Fisher Scientific, Waltham, MA USA) supplemented with 10% fetal calf serum (FCS, Biowest, Nuaillé, France), 1% MEM non-essential amino acids solution, 100 U/mL penicillin and 100 μg/mL streptomycin (all latter from Gibco/Thermo Fisher Scientific).
    Vero
    suggested: RRID:CVCL_ZW93)
    Recombinant DNA
    SentencesResources
    The sequences of the 19-amino-acids-long synthetic peptides were as follows: GRRRSVQWCAVSQPEATKC (the N-terminal pLF1, residues 1-19), EDAIWNLLRQAQEKFGKDK (the middle pLF2, residues 264-282), NLKKCSTSPLLEACEFLRK (the C-terminal pLF3, residues 674-692), and NFRTKSCPLELAKELKLCS (pCTR, a scrambled variant of pLF3).
    pLF1
    suggested: None
    pCTR
    suggested: None
    pLF3
    suggested: None

    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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  3. Version published to 10.1101/2021.12.20.473447v1 on bioRxiv