SARS‐CoV‐2 Viral Entry Proteins in Hyperandrogenemic Female Mice: Implications for Women with Polycystic Ovary Syndrome (PCOS) and COVID‐19

Huffman, Alexandra; Rezq, Samar; Basnet, Jelina; Yanes Cardozo, Licy; Romero, Damian

BACKGROUND: SARS‐CoV‐2, the causative agent of COVID‐19, infects host cells using the angiotensin I converting enzyme 2 (ACE2) as its receptor after priming by host proteases, including the transmembrane serine protease TMPRSS2. Androgen action in target tissues may partially mediate entry of SAR‐CoV‐2 and male patients have been shown to suffer increased severity and mortality compared with females. Polycystic Ovary Syndrome (PCOS) is the most common endocrine disorder in reproductive‐age women and is characterized by hyperandrogenism, ovulatory dysfunction, and polycystic ovarian morphology. PCOS is associated with obesity and cardiometabolic dysfunctions, both being risk factors associated with severe COVID‐19 pathology. The purpose of this study is to investigate the role of hyperandrogenism in a female mouse model of PCOS and to characterize the differences in host machinery required for SARS‐CoV‐2 viral entry in multiple tissues that are affected by COVID‐19 in humans. METHODS: Female mice were treated with dihydrotestosterone (DHT, 8 mg) s.c. for 90 days. Body composition was measured by EchoMRI. Fasting glucose was determined by an enzymatic method and the oral glucose tolerance test was performed with an oral glucose load (2g glucose per kg lean mass weight). mRNA and protein levels of ACE2, Tmprss2, Cathepsin L, Furin, Tmprss4, and Adam17 were quantified by RT‐qPCR, Western‐blot, and ELISA in tissues, serum, and urine. All differences are considered significant p<0.05. RESULTS: DHT treatment increased body weight, fat and lean mass. DHT treated females showed altered glucose homeostasis, having increased fasting glucose (201.10 ± 11.11 vs. 152.80 ± 9.23 mg/dL, p<0.05) and an increased area under the curve (209.2 ± 11.0 vs. 160.8 ± 3.5 mg.min/dL, p<0.05) by OGTT. The small intestine was shown to have the greatest relative Ace2 mRNA expression compared with the other tissues. Ace2 mRNA was upregulated in DHT‐treated animals in the lung (1.24‐fold), cecum (4.4‐fold), left ventricle (1.54‐fold), and kidney (2.37‐fold), and was downregulated in DHT‐treated animals in the brain (0.68‐fold) and colon (0.37‐fold). ACE2 protein expression was increased in the small intestine (1.98‐fold), left ventricle (1.30‐fold), and kidney (1.32‐fold). The SARS‐CoV‐2 priming proteases Tmprss2, Cathepsin L, and Furin mRNA were upregulated by DHT in the kidney, and protein TMPRSS2 was increased in the small intestine (1.38‐fold). ACE2 sheddase Adam17 mRNA was upregulated in the kidney by DHT, which corresponded with increased urinary ACE2 in DHT‐treated mice (14.13±3.33 vs. 0.55±0.19 pg/ng creatinine, p<0.05). CONCLUSIONS: Our results highlight the potential for increased cardiac, renal, and gastrointestinal dysfunction in PCOS women with COVID‐19. The differences observed in androgen‐treated mice highlight that several mechanisms involved in SARS‐CoV‐2entry may be altered in women with PCOS and translate to the reduced protection of specific target organs known to be susceptible to COVID‐19 injury.