HDAC1-3 inhibition increases SARS-CoV-2 replication and productive infection in lung mesothelial and epithelial cells

Flavia Trionfetti; Tonino Alonzi; Giulio Bontempi; Michela Terri; Cecilia Battistelli; Claudia Montaldo; Federica Repele; Dante Rotili; Sergio Valente; Clemens Zwergel; Giulia Matusali; Fabrizio Maggi; Delia Goletti; Marco Tripodi; Antonello Mai; Raffaele Strippoli

doi:10.3389/fcimb.2023.1257683

HDAC1-3 inhibition increases SARS-CoV-2 replication and productive infection in lung mesothelial and epithelial cells

Front Cell Infect Microbiol. 2023 Dec 13:13:1257683. doi: 10.3389/fcimb.2023.1257683. eCollection 2023.

Authors

Flavia Trionfetti^{1

2}, Tonino Alonzi³, Giulio Bontempi^{1

2}, Michela Terri², Cecilia Battistelli¹, Claudia Montaldo², Federica Repele³, Dante Rotili⁴, Sergio Valente⁴, Clemens Zwergel⁴, Giulia Matusali⁵, Fabrizio Maggi⁵, Delia Goletti³, Marco Tripodi¹, Antonello Mai^{4

6}, Raffaele Strippoli^{1

2}

Affiliations

¹ Department of Molecular Medicine, Sapienza University of Rome, Rome, Italy.
² Gene Expression Laboratory, National Institute for Infectious Diseases, Lazzaro Spallanzani IRCCS, Rome, Italy.
³ Translational Research Unit, National Institute for Infectious Diseases "Lazzaro Spallanzani"-IRCCS, Rome, Italy.
⁴ Department of Drug Chemistry and Technologies, Sapienza University of Rome, Rome, Italy.
⁵ Laboratory of Virology, National Institute for Infectious Diseases, Lazzaro Spallanzani IRCCS, Rome, Italy.
⁶ Pasteur Institute, Cenci-Bolognetti Foundation, Sapienza University of Rome, Rome, Italy.

Abstract

Background: Despite the significant progress achieved in understanding the pathology and clinical management of SARS-CoV-2 infection, still pathogenic and clinical issues need to be clarified. Treatment with modulators of epigenetic targets, i.e., epidrugs, is a current therapeutic option in several cancers and could represent an approach in the therapy of viral diseases.

Results: Aim of this study was the analysis of the role of histone deacetylase (HDAC) inhibition in the modulation of SARS-CoV-2 infection of mesothelial cells (MCs).MeT5A cells, a pleura MC line, were pre-treated with different specific class I and IIb HDAC inhibitors. Unexpectedly, treatment with HDAC1-3 inhibitors significantly increased ACE2/TMPRSS2 expression, suggesting a role in favoring SARS-CoV-2 infection. We focused our analysis on the most potent ACE2/TMPRSS2 inducer among the inhibitors analysed, MS-275, a HDAC1-3 inhibitor. ACE2/TMPRSS2 expression was validated by Western Blot (WB) and immunofluorescence. The involvement of HDAC inhibition in receptor induction was confirmed by HDAC1/HDAC2 silencing. In accordance to the ACE2/TMPRSS2 expression data, MS-275 increased SARS-CoV-2 replication and virus propagation in Vero E6 cells.Notably, MS-275 was able to increase ACE2/TMPRSS2 expression and SARS-CoV-2 production, although to a lesser extent, also in the lung adenocarcinoma cell line Calu-3 cells.Mechanistically, treatment with MS-275 increased H3 and H4 histone acetylation at ACE2/TMPRSS2 promoters, increasing their transcription.

Conclusion: This study highlights a previously unrecognized effect of HDAC1-3 inhibition in increasing SARS-CoV-2 cell entry, replication and productive infection correlating with increased expression of ACE2 and TMPRSS2. These data, while adding basic insight into COVID-19 pathogenesis, warn for the use of HDAC inhibitors in SARS-CoV-2 patients.

Keywords: ACE2 regulation; HDAC (histone deacetylase); SARS-CoV-2; TMPRSS2 expression; epidrugs; mesothelial cells; pleura; viral infection and replication.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Angiotensin-Converting Enzyme 2 / metabolism
COVID-19* / metabolism
Epithelial Cells
Histone Deacetylase 1 / metabolism
Histone Deacetylase Inhibitors / metabolism
Histone Deacetylase Inhibitors / pharmacology
Humans
Lung / metabolism
SARS-CoV-2*

Substances

entinostat
Angiotensin-Converting Enzyme 2
Histone Deacetylase Inhibitors
HDAC1 protein, human
Histone Deacetylase 1

Grants and funding

The author(s) declare financial support was received for the research, authorship, and/or publication of this article. Ministero della Salute, Ricerca Corrente ‘Linea 3’ to TA, DG, CM, FR, MTe and RS. SEED PNR 2021, Sapienza University of Rome to CB. Sapienza Università di Roma, RM12218166AEFC72 to CB. This work was supported by Ateneo Sapienza Project 2020 (RG120172B8E53D03) (Sergio Valente), and FISR2019_00374 MeDyCa (Antonello Mai). C. Zwergel is thankful for the generous funding from FSE REACT-EU within the program PON “Research and Innovation” 2014-2020, Action IV.6 “Contratti di ricerca su tematiche Green” as well as the funding from the KOHR GmbH, and the Sapienza Ateneo Project funding scheme.The research leading to some of the results reported in this work was supported by the funding from the European Union - NextGenerationEU through the Italian Ministry of University and Research under PNRR - M4C2-I1.3 Project PE_00000019 "HEAL ITALIA" to Dante Rotili (CUP B53C22004000006). The views and opinions expressed are those of the authors only and do not necessarily reflect those of the European Union or the European Commission. Kohr Aerospace was not involved in the study design, collection, analysis, interpretation of data, the writing of this article, or the decision to submit it for publication.