Skip to main page content
U.S. flag

An official website of the United States government

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2021 Sep 20;9(9):1273.
doi: 10.3390/biomedicines9091273.

Hydrogen Sulfide Inhibits TMPRSS2 in Human Airway Epithelial Cells: Implications for SARS-CoV-2 Infection

Giulia Pozzi  1 , Elena Masselli  1 , Giuliana Gobbi  1 , Prisco Mirandola  1 , Luis Taborda-Barata  2 , Luca Ampollini  1 , Paolo Carbognani  1 , Cristina Micheloni  1 , Francesco Corazza  1 , Daniela Galli  1 , Cecilia Carubbi  1 , Marco Vitale  1   3
Affiliations

Hydrogen Sulfide Inhibits TMPRSS2 in Human Airway Epithelial Cells: Implications for SARS-CoV-2 Infection

Giulia Pozzi et al. Biomedicines. .

Abstract

The COVID-19 pandemic has now affected around 190 million people worldwide, accounting for more than 4 million confirmed deaths. Besides ongoing global vaccination, finding protective and therapeutic strategies is an urgent clinical need. SARS-CoV-2 mostly infects the host organism via the respiratory system, requiring angiotensin-converting enzyme 2 (ACE2) and transmembrane protease serine 2 (TMPRSS2) to enter target cells. Therefore, these surface proteins are considered potential druggable targets. Hydrogen sulfide (H2S) is a gasotransmitter produced by several cell types and is also part of natural compounds, such as sulfurous waters that are often inhaled as low-intensity therapy and prevention in different respiratory conditions. H2S is a potent biological mediator, with anti-oxidant, anti-inflammatory, and, as more recently shown, also anti-viral activities. Considering that respiratory epithelial cells can be directly exposed to H2S by inhalation, here we tested the in vitro effects of H2S-donors on TMPRSS2 and ACE2 expression in human upper and lower airway epithelial cells. We showed that H2S significantly reduces the expression of TMPRSS2 without modifying ACE2 expression both in respiratory cell lines and primary human upper and lower airway epithelial cells. Results suggest that inhalational exposure of respiratory epithelial cells to natural H2S sources may hinder SARS-CoV-2 entry into airway epithelial cells and, consequently, potentially prevent the virus from spreading into the lower respiratory tract and the lung.

Keywords: ACE2; SARS-CoV-2; TMPRSS2; hydrogen sulfide.

PubMed Disclaimer

Conflict of interest statement

MV declares to be the Scientific Coordinator of the Italian Foundation for Research in Balneotherapy (FoRST). The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
TMPRSS2 expression in BEAS-2B and Calu-1 cells treated with NaHS. Panels (A–C): mRNA and protein expression of TMPRSS2 in BEAS-2B cells cultured without NaHS (UNT), and in presence of NaHS (0.5–1 mM) for 24, 48, 72 h. Panels (D–F): mRNA and protein expression of TMPRSS2 in Calu-1 cells cultured without NaHS (UNT), and in presence of NaHS (0.5–1 mM) for 24, 48, 72 h. In panels (A,D), TMPRSS2 mRNA expression has been normalized to rRNA18S expression for each experimental condition, and data are reported as fold increase of UNT cells for each time point. In panels (B,E), TMPRSS2 protein expression has been normalized to GAPDH expression for each experimental condition and data are reported as fold increase of UNT cells for each time point. Data are presented as means ± SD of at least 3 independent experiments (*, p < 0.05; **, p < 0.01; ***, p < 0.001; ****, p < 0.0001; ## vs. NaHS 0.5 mM 72 h, p < 0.01; One-way ANOVA and Dunnett’s test). Panels (C,F): representative blots obtained with BEAS-2B and Calu-1 cells, respectively.
Figure 2
Figure 2
TMPRSS2 expression in BEAS-2B and Calu-1 cells treated with GYY4137. Panels (A–C): mRNA and protein expression of TMPRSS2 in BEAS-2B cells cultured without GYY4137 (DMSO) and in presence GYY4137 (0.05–0.1 mM) for 24, 48, 72 h. Panels (D–F): mRNA and protein expression of TMPRSS2 in Calu-1 cells cultured without GYY4137 (DMSO) and in presence GYY4137 (0.05–0.1 mM) for 24, 48, 72 h. In panels (A,D), TMPRSS2 mRNA expression has been normalized to rRNA18S expression for each experimental condition, and data are reported as fold increase of DMSO treated cells for each time point. In panels (B,E), TMPRSS2 protein expression has been normalized to GAPDH expression for each experimental condition and data are reported as fold increase of DMSO treated cells for each time point. Data are presented as mean ± SD of at least 3 independent experiments (*, p < 0.05; **, p <0.01; ***, p < 0.001; One-way ANOVA and Dunnett’s test). Panels (C,F): representative blots obtained with BEAS-2B and Calu-1 cells, respectively.
Figure 3
Figure 3
TMPRSS2 expression in HNEpC. Panels (A–C): mRNA and protein expression of TMPRSS2 in HNEpC cultured without NaHS (UNT), and in presence of NaHS (0.5–1 mM) for 24, 48 h. In panel (A), TMPRSS2 mRNA expression has been normalized to rRNA18S expression for each experimental condition, and data are reported as fold increase of UNT treated cells for each time point. In panel (B), TMPRSS2 protein expression has been normalized to GAPDH expression for each experimental condition and data are reported as fold increase of DMSO treated cells for each time point. Data are presented as mean ± SD of at least 3 independent experiments (*, p < 0.05; ***, p < 0.0001 One-way ANOVA and Dunnett’s test). Panel (C): representative blot obtained with HNEpC. Panel (D): immunohistochemistry analysis of TMPRSS2 in representative HNEpC cultured without NaHS (UNT), and in presence of NaHS (0.5–1 mM) for 24, 48 h; scale bar = 100 µM.
Figure 4
Figure 4
TMPRSS2 expression in the lung. Immunohistochemistry analysis of TMPRSS2 in a representative lung specimen incubated in RPMI medium without NaHS (UNT), and in presence of NaHS (0.5–1 mM) for 24, 48 h; scale bar = 50 µM.
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

See all "Cited by" articles

References

    1. Li F. Structure, function, and evolution of coronavirus spike proteins. Annu. Rev. Virol. 2016;3:237–261. doi: 10.1146/annurev-virology-110615-042301. - DOI - PMC - PubMed
    1. He Y., Zhou Y., Liu S., Kou Z., Li W., Farzan M., Jiang S. Receptor-binding domain of SARS-CoV spike protein induces highly potent neutralizing antibodies: Implication for developing subunit vaccine. Biochem. Biophys. Res. Commun. 2004;324:773–781. doi: 10.1016/j.bbrc.2004.09.106. - DOI - PMC - PubMed
    1. Hoffmann M., Kleine-Weber H., Schroeder S., Krüger N., Herrler T., Erichsen S., Schiergens T.S., Herrler G., Wu N.H., Nitsche A., et al. SARS-CoV-2 cell entry depends on ACE2 and TMPRSS2 and is blocked by a clinically proven protease inhibitor. Cell. 2020;181:271–280.e8. doi: 10.1016/j.cell.2020.02.052. - DOI - PMC - PubMed
    1. Zhou P., Yang X.-L., Wang X.-G., Hu B., Zhang L., Zhang W., Si H.-R., Zhu Y., Li B., Huang C.-L., et al. A pneumonia outbreak associated with a new coronavirus of probable bat origin. Nature. 2020;579:270–273. doi: 10.1038/s41586-020-2012-7. - DOI - PMC - PubMed
    1. Walls A.C., Park Y., Tortorici M.A., Wall A., McGuire A.T., Veesler D. Structure, function, and antigenicity of the SARS-CoV-2 spike glycoprotein. Cell. 2020;181:281–292.e6. doi: 10.1016/j.cell.2020.02.058. - DOI - PMC - PubMed

LinkOut - more resources