Topical TMPRSS2 inhibition prevents SARS-CoV-2 infection in differentiated primary human airway cells
This article has been Reviewed by the following groups
Discuss this preprint
Start a discussion What are Sciety discussions?Listed in
- Evaluated articles (ScreenIT)
Abstract
Background
There are limited effective prophylactic treatments for SARS-CoV-2 infection, and limited early treatment options. Viral cell entry requires spike protein binding to the ACE2 receptor and spike cleavage by TMPRSS2, a cell surface serine protease. Targeting of TMPRSS2 by either androgen blockade or direct inhibition is already in clinical trials in early SARS-CoV-2 infection.
Methods
The likely initial cells of SARS-CoV-2 entry are the ciliated cells of the upper airway. We therefore used differentiated primary human airway epithelial cells maintained at the air-liquid interface (ALI) 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, camostat is rapidly metabolised in the circulation in vivo , and systemic bioavailability after oral dosing is low. We therefore modelled local airway administration by applying camostat to the apical surface of the differentiated ALI cultures. We demonstrated that a brief exposure to topical camostat is effective at restricting SARS-CoV-2 viral 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.
Article activity feed
-
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.
-
