OR02-07 siRNA Targeting Hepatic Androgen Receptor (AR) Ameliorates Gluconeogenesis and Insulin Resistance in PCOS Mouse Models

Abstract Disclosure: M.A. Rahman: None. B. Garfinkel: None. S. Andrisse: None. Hyperandrogenism in women of reproductive age is associated with polycystic ovary syndrome (PCOS) and metabolic disorders such as obesity, diabetes and nonalcoholic fatty liver diseases. Globally PCOS affects nearly 10% of women of reproductive age with limited treatment options. Evidence suggests that genetic knockout of hepatocyte androgen receptor (AR) prevented PCOS related glucose dysregulation in female mice treated with low-dose dihydrotestosterone (DHT). The interconnectivity of hepatic AR, metabolic dysfunction and PCOS can be explained, in part, by the hepatic insulin resistance (HIR) and increased gluconeogenesis fueled by hyperandrogenemia. Considering the limited targeted treatment options for PCOS and hyperandrogenemia, hepatic AR targeting may provide an innovative therapeutic approach. As such, we investigated the efficacy of a small interfering RNA (siRNA) targeting hepatic androgen receptor (AR) in female mice treated with 33 μg/d letrozole (LET), an aromatase inhibitor. Twenty-two 4-week-old female C57BL/6J mice were randomly assigned into three groups and subcutaneously implanted with continuous-release pellets containing LET pellets (n=6) or placebo control pellets (CON) (n=8). The third group (n=8) was implanted with LET pellets and administered with AR-siRNA (10 mg/kg body weight) (LET+siR) subcutaneously for every two weeks. Mice were weighed weekly and metabolic tests were performed until sacrifice. RT-qPCR analysis revealed that specific siRNAs directed against AR led to a selective knock-down of the hepatic AR expression by about 60-70% in the treated mice group as compared to the LET-only group after 120 days of treatment. Metabolic tests such as Glucose Tolerance Test (GTT) and Insulin Tolerance Test (ITT) revealed no significant differences in systemic insulin resistance across groups; however, the LET group of mice exhibited significantly impaired pyruvate tolerance ((at 60 min, p = 0.0026 and at 90 min, p = 0.0007 ) , which was ameliorated in LET+siRNA mice to levels comparable to controls. These findings also paralleled with a reduction of genes involved in the upregulation of hepatic gluconeogenesis. Western Blot analysis suggests that insulin signaling intermediates (p-AKT and p-Foxo1) were rescued from LET-induced HIR in the LET+siRNA group. Taken together, the findings suggest that: 1) the current letrozole dose (33 ug/day) induces HIR rather than a systemic metabolic disorder; and 2) treatment with the siRNA targeting the hepatic androgen receptor can ameliorate HIR induced by LET. Altogether, our findings highlighted that targeting the hepatic AR using siRNA warrants further exploration as a potential treatment for PCOS. Presentation: Saturday, July 12, 2025