Topical TMPRSS2 inhibition prevents SARS-CoV-2 infection in differentiated human airway cultures
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Abstract
Background: There are limited effective prophylactic/early treatments for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection. Viral entry requires spike protein binding to the angiotensin-converting enzyme-2 receptor and cleavage by transmembrane serine protease 2 (TMPRSS2), a cell surface serine protease. Targeting of TMPRSS2 by either androgen blockade or direct inhibition is in clinical trials in early SARS-CoV-2 infection. Methods: We used differentiated primary human airway epithelial cells at the air–liquid interface to test the impact of targeting TMPRSS2 on the prevention of SARS-CoV-2 infection. Results: We first modelled the systemic delivery of compounds. Enzalutamide, an oral androgen receptor antagonist, had no impact on SARS-CoV-2 infection. By contrast, camostat mesylate, an orally available serine protease inhibitor, blocked SARS-CoV-2 entry. However, oral camostat is rapidly metabolised in the circulation, with poor airway bioavailability. We therefore modelled local airway administration by applying camostat to the apical surface of differentiated airway cultures. We demonstrated that a brief exposure to topical camostat effectively restricts SARS-CoV-2 infection. Conclusion: These experiments demonstrate a potential therapeutic role for topical camostat for pre- or post-exposure prophylaxis of SARS-CoV-2, which can now be evaluated in a clinical trial.
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SciScore for 10.1101/2021.04.23.440619: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Ethics IRB: model: Human airway epithelial cells (hAECs) were purchased from Lonza or were expanded directly from either bronchial brushings from a main airway at bronchoscopy or a nasal brushing from the inferior turbinate from patients at Cambridge University Hospitals NHS Trust (Research Ethics Committee Reference 19/SW/0152). Sex as a biological variable Specific cells used were human bronchial epithelial cells (HBECs) derived from a non-smoking donor (Lonza; Cat# CC-2540, male); human bronchial epithelial cells from a male smoking donor undergoing bronchoscopy for a non-cancer indication, and nasal epithelial cells (MOD006) from a male patient. Randomization not detected. Blinding not detected. Power … SciScore for 10.1101/2021.04.23.440619: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Ethics IRB: model: Human airway epithelial cells (hAECs) were purchased from Lonza or were expanded directly from either bronchial brushings from a main airway at bronchoscopy or a nasal brushing from the inferior turbinate from patients at Cambridge University Hospitals NHS Trust (Research Ethics Committee Reference 19/SW/0152). Sex as a biological variable Specific cells used were human bronchial epithelial cells (HBECs) derived from a non-smoking donor (Lonza; Cat# CC-2540, male); human bronchial epithelial cells from a male smoking donor undergoing bronchoscopy for a non-cancer indication, and nasal epithelial cells (MOD006) from a male patient. Randomization not detected. Blinding not detected. Power Analysis not detected. Cell Line Authentication not detected. Table 2: Resources
Antibodies Sentences Resources Permeabilised cells were pelleted, stained for 15 minutes at room temperature in 100 μL of sheep anti-SARS-CoV-2 nucleoprotein antibody (MRC-PPU, DA114) at a concentration of 0.7 μg/mL, washed and incubated in 100 μL AF488 donkey anti-sheep (Jackson ImmunoResearch #713-545-147) at a concentration of 2 μg/mL for 15 minutes at room temperature. anti-SARS-CoV-2 nucleoproteinsuggested: Noneanti-sheepsuggested: (Jackson ImmunoResearch Labs Cat# 713-545-147, RRID:AB_2340745)The following antibodies were used: Anti-ACE2 antibody (Anti-ACE2 antibody - N-terminal, ab228349 was originally used. Anti-ACE2suggested: NoneExperimental Models: Cell Lines Sentences Resources Viral titre was determined by 50% tissue culture infectious dose (TCID50) in Huh7-ACE2 cells. Huh7-ACE2suggested: NoneSoftware and Algorithms Sentences Resources % Cytotoxicity was calculated: Quantification and statistical analysis: Statistical analyses of mRNA expression assays and infection quantification data were performed using Prism 8 software (GraphPad Software).
Prismsuggested: (PRISM, RRID:SCR_005375)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: We found the following clinical trial numbers in your paper:
Identifier Status Title NCT04446429 Completed Anti-Androgen Treatment for COVID-19 NCT04475601 Recruiting Enzalutamide Treatment in COVID-19 NCT04455815 Recruiting A Trial Looking at the Use of Camostat to Reduce Progression… NCT04608266 Recruiting CAMOVID : Evaluation of Efficacy and Safety of Camostat Mesy… Results from Barzooka: We did not find any issues relating to the usage of bar graphs.
Results from JetFighter: Please consider improving the rainbow (“jet”) colormap(s) used on pages 32, 29, 30 and 31. At least one figure is not accessible to readers with colorblindness and/or is not true to the data, i.e. not perceptually uniform.
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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