Cardiac and Renal SARS-CoV-2 Viral Entry Protein Regulation by Androgens and Diet: Implications for Polycystic Ovary Syndrome and COVID-19

Figures

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  Figure 1. Effect of DHT and diet on body weight and composition, and cardiac and renal hypertrophy. Animals were treated with dihydrotestosterone (DHT) or vehicle (Veh) and maintained in low (LFD) or high (HFD) fat diet for 90 days. (A) Body weight was determined by gravimetry. (B) Fat mass was determined by EchoMRI. Left ventricle (LV) (C) and kidney (D) weights were determined by gravimetry and corrected by tibia length (TL). Data are expressed as mean ± SEM. N = 8/group. Data were analyzed by two-way ANOVA followed by Fisher’s LSD test. a p < 0.05 vs. LFD-Veh; b p < 0.05 vs. HFD-Veh; c p < 0.05 vs. LFD-DHT.

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  Figure 2. Effect of DHT and diet on androgen receptor mRNA and protein expression. Animals were treated with dihydrotestosterone (DHT) or vehicle (Veh) and maintained in low (LFD) or high (HFD) fat diet for 90 days. Left ventricle (LV) (A) and kidney (B) Androgen receptor (AR) mRNA was quantified by RT-qPCR (N = 6–8/group). LV (C) and kidney (D) AR protein was quantified by Western-blot (N = 4/group). Data are expressed as mean ± SEM. Data were analyzed by two-way ANOVA followed by Fisher’s LSD test. a p < 0.05 vs. LFD-Veh; b p < 0.05 vs. HFD-Veh; c p < 0.05 vs. LFD-DHT.

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  Figure 3. Effect of DHT and diet on cardiac SARS-CoV-2 viral entry proteins expression. Animals were treated with dihydrotestosterone (DHT) or vehicle (Veh) and maintained in low (LFD) or high (HFD) fat diet for 90 days. Ace2 (A), Tmprss2 (B), furin (C), cathepsin L (D), and ADAM17 (E) mRNA was quantified by RT-qPCR and standardized to the geometric mean of four housekeeping genes (HGMK) (N = 6/group). ACE2 (F), TMPRSS2 (G), furin (H), cathepsin L (I), and ADAM17 (J) protein was quantified by Western-blot and normalized to GAPDH (N = 4/group). Total furin (H) was quantified as the sum of profurin (dashed arrow) and cleaved soluble furin (solid arrow). Data are expressed as mean ± SEM. Data were analyzed by two-way ANOVA followed by Fisher’s LSD test. a p < 0.05 vs. LFD-Veh; b p < 0.05 vs. HFD-Veh; c p < 0.05 vs. LFD-DHT.

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  Figure 4. Effect of DHT and diet on renal SARS-CoV-2 viral entry proteins expression. Animals were treated with dihydrotestosterone (DHT) or vehicle (Veh) and maintained in low (LFD) or high (HFD) fat diet for 90 days. Ace2 (A), Tmprss2 (B), Tmprss4 (C), furin (D), cathepsin L (E), and ADAM17 (F) mRNA was quantified by RT-qPCR and standardized to the geometric mean of four housekeeping genes (HGMK) (N = 6–8/group). ACE2 (G), TMPRSS2 (H), TMPRSS4 (I), furin (J), cathepsin L (K), and ADAM17 (L) protein was quantified by Western-blot and normalized to GAPDH (N = 4/group). Total furin (J) was quantified as the sum of profurin (dashed arrow) and cleaved soluble furin (solid arrow). Data are expressed as mean ± SEM. Data were analyzed by two-way ANOVA followed by Fisher’s LSD test. a p < 0.05 vs. LFD-Veh; b p < 0.05 vs. LFD-Veh; c p < 0.05 vs. LFD-DHT.

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  Figure 5. Effect of DHT and diet on serum and urinary ACE2. Animals were treated with dihydrotestosterone (DHT) or vehicle (Veh) and maintained in low (LFD) or high (HFD) fat diet for 90 days. Serum (A, N = 4–7/group) and urinary (B, N = 4–6/group) ACE2 were quantified by ELISA. Urinary ACE2 was corrected by urinary creatinine. Data are expressed as mean ± SEM. Data were analyzed by two-way ANOVA followed by Fisher’s LSD test. a p < 0.05 vs. LFD-Veh; b p < 0.05 vs. HFD-Veh.

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  Figure 6. Correlations between renal androgen receptor mRNA expression and SARS-CoV-2 viral entry proteins. Animals were treated with dihydrotestosterone (DHT) or vehicle (Veh) and maintained in low (LFD) or high (HFD) fat diet for 90 days. Androgen receptor and SARS-CoV-2 viral entry proteins renal mRNA expression was quantified by RT-qPCR. Pearson’s correlation coefficients (r) were calculated between androgen receptor and Ace2 (A), Tmprss2 (B), Tmprss4 (C), furin (D), cathepsin L (E), and ADAM17 (F) mRNA expression.

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  Figure 7. Proposed mechanisms for increased risk for severe COVID-19 cardiorenal outcomes in normal women and women with PCOS. Top panels: Women with PCOS have increased body weight, fat mass, and cardiac and renal hypertrophy. Lower panels: Androgens differentially regulate the cardiac and renal expression of SARS-CoV-2 viral entry proteins including the SARS-CoV-2 receptor ACE2 and several cellular proteases that promote viral entry and propagation. HFD exacerbates androgens-mediated obesity and cardiorenal hypertrophic response and modulates androgens’ effect on the expression of the different cellular proteases. The red stars indicates the parameters that are exacerbated with HFD.