Conversion of Classical and 11-Oxygenated Androgens by Insulin-Induced AKR1C3 in a Model of Human PCOS Adipocytes

Polycystic ovary syndrome (PCOS) is the most prevalent endocrinopathy in women. A common symptom of PCOS is hyperandrogenism (AE); however, the source of these androgens is uncertain. Aldo-keto reductase family 1 member C3 (AKR1C3) catalyzes the formation of testosterone (T) and 5 alpha-dihydrotestosterone (DHT) in peripheral tissues, which activate the androgen receptor (AR). AKR1C3 is induced by insulin in adipocytes and may be central in driving the AE in PCOS. We elucidated the conversion of both classical and 11-oxygenated androgens to potent androgens in a model of PCOS adipocytes. Using high-performance liquid chromatography (HPLC) discontinuous kinetic assays to measure product formation by recombinant AKR1C3, we found that the conversion of 11-keto-Delta 4-androstene-3,17-dione (11K-4AD) to 11-ketotestosterone (11K-T) and 11-keto-5 alpha-androstane-3,17-dione (11K-5AD) to 11-keto-5 alpha-dihydrotestosterone (11K-DHT) were superior to the formation of T and DHT. We utilized a stable isotope dilution liquid chromatography high resolution mass spectrometric (SID-LC-HRMS) assay for the quantification of both classical and 11-oxygenated androgens in differentiated Simpson-Golabi-Behmel syndrome adipocytes in which AKR1C3 was induced by insulin. Adipocytes were treated with adrenal derived 11 beta-hydroxy-Delta 4-androstene-3,17-dione (11 beta-OH-4AD), 11K-4AD, or Delta 4-androstene-3,17-dione (4AD). The conversion of 11 beta-OH-4AD and 11K-4AD to 11K-T required AKR1C3. We also found that once 11K-T is formed, it is inactivated to 11 beta-hydroxy-testosterone (11 beta-OH-T) by 11 beta-hydroxysteroid dehydrogenase type 1 (HSD11B1). Our data reveal a unique role for HSD11B1 in protecting the AR from AE. We conclude that the 11-oxygenated androgens formed in adipocytes may contribute to the hyperandrogenic profile of PCOS women and that AKR1C3 is a potential therapeutic target to mitigate the AE of PCOS.

Automated summary

Polycystic ovary syndrome (PCOS) is a common endocrine disorder in women, characterized by hyperandrogenism. Research suggests that Aldo-keto reductase family 1 member C3 (AKR1C3) plays a significant role in the conversion of androgens in adipocytes, contributing to the hyperandrogenic profile of PCOS. This finding highlights AKR1C3 as a potential therapeutic target for mitigating the symptoms of PCOS, particularly through the modulation of 11-oxygenated androgens formed in adipocytes.