Androgen regulation of pulmonary AR, TMPRSS2 and ACE2 with implications for sex-discordant COVID-19 outcomes

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Abstract

The sex discordance in COVID-19 outcomes has been widely recognized, with males generally faring worse than females and a potential link to sex steroids. A plausible mechanism is androgen-induced expression of TMPRSS2 and/or ACE2 in pulmonary tissues that may increase susceptibility or severity in males. This hypothesis is the subject of several clinical trials of anti-androgen therapies around the world. Here, we investigated the sex-associated TMPRSS2 and ACE2 expression in human and mouse lungs and interrogated the possibility of pharmacologic modification of their expression with anti-androgens. We found no evidence for increased TMPRSS2 expression in the lungs of males compared to females in humans or mice. Furthermore, in male mice, treatment with the androgen receptor antagonist enzalutamide did not decrease pulmonary TMPRSS2. On the other hand, ACE2 and AR expression was sexually dimorphic and higher in males than females. ACE2 was moderately suppressible with enzalutamide administration. Our work suggests that sex differences in COVID-19 outcomes attributable to viral entry are independent of TMPRSS2. Modest changes in ACE2 could account for some of the sex discordance.

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  1. SciScore for 10.1101/2020.04.21.051201: (What is this?)

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

    Table 1: Rigor

    Institutional Review Board StatementIACUC: Study approval: Mouse studies were performed under a protocol approved by the Institutional Animal Care and Use Committee (IACUC) of the Cleveland Clinic Lerner Research Institute.
    Randomizationnot detected.
    Blindingnot detected.
    Power Analysisnot detected.
    Sex as a biological variableThe male mice were arbitrarily divided between two groups receiving control chow or Enz diet 62.5 mg/kg.
    Cell Line Authenticationnot detected.

    Table 2: Resources

    Antibodies
    SentencesResources
    The lysates were then centrifuged for 15 min at 16,000 x g and the supernatants were collected for immunoblot analysis with antibodies for TMPRSS2 (Abcam: ab92323 and Proteintech: 14437-1-AP), AR (EMD Millipore: PG-21 and Santa Cruz Biotechnology: N-20), PSA (Cell signaling: D6B1) and GAPDH (D16H11).
    TMPRSS2
    suggested: (Abcam Cat# ab56111, RRID:AB_883079)
    GAPDH
    suggested: (Cell Signaling Technology Cat# 5174, RRID:AB_10622025)
    The antibodies were visualized using the OmniMap anti-Rabbit HRP (05269679001; Roche), and OmniMap anti-Goat HRP (06607233001; Roche) in conjunction with the ChromoMap DAB detection kit (05266645001; Roche).
    anti-Rabbit
    suggested: None
    HRP
    suggested: None
    Experimental Models: Cell Lines
    SentencesResources
    ChIP-seq: ChIP-Seq analysis was performed in LNCaP cells treated with vehicle (DMSO) or Enz 10 μM.
    LNCaP
    suggested: None
    Experimental Models: Organisms/Strains
    SentencesResources
    Mice, treatments and lung harvest: A cohort of adult NSG mice (> 6 weeks old) were obtained from Cleveland Clinic Biological Resources Unit.
    NSG
    suggested: None
    Software and Algorithms
    SentencesResources
    Gene expression in human lung: The public genomics data repository Gene Expression Omnibus (GEO, ncbi.nlm.nih.gov/geo) was searched for data sets containing expression profiling of samples from non-cancerous human lung and bronchial/airway epithelial cells with samples identified by gender and/or smoking status of subjects.
    Gene Expression Omnibus
    suggested: (Gene Expression Omnibus (GEO, RRID:SCR_005012)
    Analyses were performed in R or GraphPad Prism.
    GraphPad Prism
    suggested: (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:

    IdentifierStatusTitle
    NCT04374279Not yet recruitingTrial to Promote Recovery From COVID-19 With Endocrine Thera…
    NCT04475601RecruitingEnzalutamide Treatment in COVID-19
    NCT04509999RecruitingBicalutamide to Block TMPRSS2 in Males With COVID-19 Infecti…
    NCT04397718RecruitingHormonal Intervention for the Treatment in Veterans With COV…
    NCT045009999Trial number did not resolve on clinicaltrials.gov. Is the number correct?NA


    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

    SciScore is an automated tool that is designed to assist expert reviewers by finding and presenting formulaic information scattered throughout a paper in a standard, easy to digest format. SciScore checks for the presence and correctness of RRIDs (research resource identifiers), and for rigor criteria such as sex and investigator blinding. For details on the theoretical underpinning of rigor criteria and the tools shown here, including references cited, please follow this link.