DNA methylation and gene expression pattern of ACE2 and TMPRSS2 genes in saliva samples of patients with SARS-CoV-2 infection

  1. Pratibha Misra
  2. Bhasker Mukherjee
  3. Rakhi Negi
  4. Vikas Marwah
  5. Arijit Kumar Ghosh
  6. Prashant Jindamwar
  7. Mukesh U Singh
  8. Y Vashum
  9. R Syamraj
  10. G Bala Chandra
  11. M K Sibin

Reviewed by

Read the full article See related articles

Listed in

Log in to save this article

Abstract

COVID-19 caused by SARS-CoV-2 became a pandemic affecting the health and economy of the world. Although it was known that this virus uses ACE2 protein along with TMPRSS2 to enter the host cell, the methylation pattern and gene expression of ACE2 and TMPRSS2 genes are not explored in saliva samples of patients infected with COVID-19. The study aimed to quantify promoter methylation of ACE2 and TMPRSS2 along with its mRNA expression in saliva samples of COVID-19 patients in order to understand the regulatory mechanism of these genes in SARS-CoV-2 infection. Saliva samples were collected from thirty male patients with SARS-CoV-2 infection and thirty age-matched healthy control male subjects. Q MS PCR and qRT PCR was performed to quantify the promoter DNA methylation and mRNA expression of ACE2 and TMPRSS2 respectively. Our study didn’t find any significant difference between methylation and expression of these two genes in cases compared to control subjects. However there was significant positive correlation between DNA methylation of ACE2 and its gene expression. Among cases, the sample collected ≥7 days after appearance of symptoms showed higher amount of methylation in both ACE2 and TMPRSS2 genes when compared to sample collected before 7 days. In conclusion, we found that ACE2 and TMPRSS2 methylation plays a role in COVID-19.

Article activity feed

  1. ScreenIT

    SciScore for 10.1101/2020.10.24.20218727: (What is this?)

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

    Table 1: Rigor

    Institutional Review Board StatementIACUC: Ethical clearance was taken from the Institutional Ethics Committee, AFMC (S No. IEC/2020/95).
    Consent: After obtaining written informed consent, 30 random subjects who were tested positive for SARS-CoV-2 infection by a reverse transcription-polymerase chain reaction (RT-PCR) were recruited in the study.
    Randomizationnot detected.
    Blindingnot detected.
    Power Analysisnot detected.
    Sex as a biological variablenot detected.

    Table 2: Resources

    Software and Algorithms
    SentencesResources
    Primers were designed using Gene Runner software (Version 3.05).
    Gene Runner
    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: We detected the following sentences addressing limitations in the study:
    Limitations of our study include small sample size and exclusion of severe cases. Saliva samples may contain white blood cells which may contribute DNA that can be a confounder [Theda et al, 2018]. In future, a follow up study with patients with SARS-CoV-2 during infection and after recovery will give more insight on host epigenetic adaptation. A study with mild/moderate versus severe cases may delineate the pathological role of ACE2/ TMPRSS2 expression in severe cases.

    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.

    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.

  2. Version published to 10.1101/2020.10.24.20218727 on medRxiv