Alpha 1 Antitrypsin is an Inhibitor of the SARS-CoV-2–Priming Protease TMPRSS2
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Abstract
Background: Host proteases have been suggested to be crucial for dissemination of MERS, SARS-CoV, and SARS-CoV-2 coronaviruses, but the relative contribution of membrane versus intracellular proteases remains controversial. Transmembrane serine protease 2 (TMPRSS2) is regarded as one of the main proteases implicated in the coronavirus S protein priming, an important step for binding of the S protein to the angiotensin-converting enzyme 2 (ACE2) receptor before cell entry. Methods: We developed a cell-based assay to identify TMPRSS2 inhibitors. Inhibitory activity was established in SARS-CoV-2 viral load systems. Results: We identified the human extracellular serine protease inhibitor (serpin) alpha 1 antitrypsin (A1AT) as a novel TMPRSS2 inhibitor. Structural modeling revealed that A1AT docked to an extracellular domain of TMPRSS2 in a conformation that is suitable for catalysis, resembling similar serine protease inhibitor complexes. Inhibitory activity of A1AT was established in a SARS-CoV-2 viral load system. Notably, plasma A1AT levels were associated with COVID-19 disease severity. Conclusions: Our data support the key role of extracellular serine proteases in SARS CoV-2 infections and indicate that treatment with serpins, particularly the FDA-approved drug A1AT, may be effective in limiting SARS-CoV-2 dissemination by affecting the surface of the host cells.
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SciScore for 10.1101/2020.05.04.077826: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Institutional Review Board Statement IRB: COVID-19 plasma analysis: Adults who presented to the University of Cincinnati Medical Center (UCMC) Emergency Department (ED) with suspected COVID-19 and had a clinically indicated blood draw were prospectively enrolled via an institutional review board–approved waiver of informed consent.
Consent: COVID-19 plasma analysis: Adults who presented to the University of Cincinnati Medical Center (UCMC) Emergency Department (ED) with suspected COVID-19 and had a clinically indicated blood draw were prospectively enrolled via an institutional review board–approved waiver of informed consent.Randomization not detected. Blinding not detected. Power Analysis not detected. S… SciScore for 10.1101/2020.05.04.077826: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Institutional Review Board Statement IRB: COVID-19 plasma analysis: Adults who presented to the University of Cincinnati Medical Center (UCMC) Emergency Department (ED) with suspected COVID-19 and had a clinically indicated blood draw were prospectively enrolled via an institutional review board–approved waiver of informed consent.
Consent: COVID-19 plasma analysis: Adults who presented to the University of Cincinnati Medical Center (UCMC) Emergency Department (ED) with suspected COVID-19 and had a clinically indicated blood draw were prospectively enrolled via an institutional review board–approved waiver of informed consent.Randomization not detected. Blinding not detected. Power Analysis not detected. Sex as a biological variable not detected. Cell Line Authentication not detected. Table 2: Resources
Antibodies Sentences Resources Gels were transferred to nitrocellulose membranes (Life Technologies) and probed with the primary antibodies rabbit anti-V5 (Bethyl Laboratories), mouse anti-TMPRSS2 (Santa Cruz), and rabbit anti–human GAPDH (ABCAM) and subsequently with the secondary antibody IRDye 800RD goat anti-rabbit and IRDye 680RD goat anti-mouse (LI-COR Biosciences). anti-V5suggested: Noneanti-TMPRSS2suggested: Noneanti–human GAPDHsuggested: Noneanti-rabbitsuggested: (LI-COR Biosciences Cat# 925-68073, RRID:AB_2716687)anti-mousesuggested: NoneExperimental Models: Cell Lines Sentences Resources HEK-293T cell culture and transfection: HEK-293T cells were grown in Dulbecco’s modified eagle media (DMEM) supplemented with 10% fetal bovine serum (FBS) and seeded in a black, 96-well plate (75,000 cells/well). HEK-293Tsuggested: NoneCaco-2 cells were plated in 96-well plates. Caco-2suggested: NoneSoftware and Algorithms Sentences Resources Statistical analyses were performed using GraphPad Prism (GraphPad Software Incorporated). GraphPad Prismsuggested: (GraphPad Prism, RRID:SCR_002798)GraphPadsuggested: (GraphPad Prism, RRID:SCR_002798)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.
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