SARS-CoV-2 Enters Human Leydig Cells and Affects Testosterone Production In Vitro

Lu Li; Chantal M Sottas; Hsu-Yu Chen; Yuchang Li; Haoyi Cui; Jason S Villano; Joseph L Mankowski; Paula M Cannon; Vassilios Papadopoulos

doi:10.3390/cells12081198

SARS-CoV-2 Enters Human Leydig Cells and Affects Testosterone Production In Vitro

Cells. 2023 Apr 20;12(8):1198. doi: 10.3390/cells12081198.

Authors

Lu Li¹, Chantal M Sottas¹, Hsu-Yu Chen², Yuchang Li¹, Haoyi Cui¹, Jason S Villano³, Joseph L Mankowski³, Paula M Cannon², Vassilios Papadopoulos¹

Affiliations

¹ Department of Pharmacology and Pharmaceutical Sciences, Alfred E. Mann School of Pharmacy and Pharmaceutical Sciences, University of Southern California, Los Angeles, CA 90089, USA.
² Department of Molecular Microbiology and Immunology, Keck School of Medicine, University of Southern California, Los Angeles, CA 90033, USA.
³ Departments of Molecular and Comparative Pathobiology, Pathology and Neurology, The Johns Hopkins School of Medicine, Baltimore, MD 21205, USA.

Abstract

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), a SARS-like coronavirus, continues to produce mounting infections and fatalities all over the world. Recent data point to SARS-CoV-2 viral infections in the human testis. As low testosterone levels are associated with SARS-CoV-2 viral infections in males and human Leydig cells are the main source of testosterone, we hypothesized that SARS-CoV-2 could infect human Leydig cells and impair their function. We successfully detected SARS-CoV-2 nucleocapsid in testicular Leydig cells of SARS-CoV-2-infected hamsters, providing evidence that Leydig cells can be infected with SARS-CoV-2. We then employed human Leydig-like cells (hLLCs) to show that the SARS-CoV-2 receptor angiotensin-converting enzyme 2 is highly expressed in hLLCs. Using a cell binding assay and a SARS-CoV-2 spike-pseudotyped viral vector (SARS-CoV-2 spike pseudovector), we showed that SARS-CoV-2 could enter hLLCs and increase testosterone production by hLLCs. We further combined the SARS-CoV-2 spike pseudovector system with pseudovector-based inhibition assays to show that SARS-CoV-2 enters hLLCs through pathways distinct from those of monkey kidney Vero E6 cells, a typical model used to study SARS-CoV-2 entry mechanisms. We finally revealed that neuropilin-1 and cathepsin B/L are expressed in hLLCs and human testes, raising the possibility that SARS-CoV-2 may enter hLLCs through these receptors or proteases. In conclusion, our study shows that SARS-CoV-2 can enter hLLCs through a distinct pathway and alter testosterone production.

Keywords: ACE2; Leydig cells; SARS-CoV-2; SARS-CoV-2 spike pseudovectors; TMPRSS2; cinanserin; hamster; human; spike protein; testis; testosterone.

Publication types

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

MeSH terms

COVID-19* / metabolism
Humans
Leydig Cells / metabolism
Male
Peptidyl-Dipeptidase A / metabolism
SARS-CoV-2* / metabolism
Testis / metabolism
Testosterone / metabolism

Substances

Testosterone
Peptidyl-Dipeptidase A

Grants and funding

This work was supported by funds from the USC Mann School of Pharmacy and Pharmaceutical Sciences and the John Stauffer Decanal Chair in Pharmaceutical Sciences (USC) to V.P., a grant to P.M.C. from the W.M. Keck Foundation COVID-19 Research Fund to the Keck School of Medicine of USC, and the generosity of the collective community of donors to the Johns Hopkins University School of Medicine for COVID research (J.L.M.).