SAT-019 Clinical Disorders and Diseases Marked by Elevated Adrenal C11-oxy C(21) and C11-oxy C(19) Steroid Levels

Adrenal steroid hormones have been implicated in numerous clinical conditions with adrenal androgens marked as major contributors in the progression of androgen-dependent diseases and disorders such as congenital adrenal hyperplasia, 21-hydroxylase deficiency, polycystic ovary syndrome (PCOS) and castration-resistant prostate cancer. Although adrenal C(19) androgens, androstenedione and testosterone, have been routinely monitored in patients under clinical care, it is only recently that the contribution of the C11-oxy androgens has come to the attention of clinicians. Adrenal 11β-hydroxyandrostenedione (11OHA4) is systemically converted by steroidogenic enzymes to produce potent C11-oxy C(19) steroids, notably 11keto-testosterone and 11keto-dihydrotestosterone (11KDHT) in the 11OHA4-pathway. Furthermore, we recently identified adrenal C11-oxy C(21) steroids as potential contributors to androgen excess via the C11-oxy C(21) backdoor pathway −11β-hydroxyprogesterone and 11keto-progesterone (11KPROG), together with 21-deoxycortisol and 21-deoxycortisone are converted to 11KDHT, thus potentially adding to the androgen pool in disease states. Using a high-throughput ultra performance convergence chromatography tandem mass spectrometry (UPC(2)-MS/MS) method for the separation and quantification of C11-oxy steroids and their downstream metabolites, we were able to simultaneously analyze 51 steroids: 31 C(21) and 20 C(19), which include the C11-oxy C(21) and C11-oxy C(19) steroids, in a single 8 min chromatographic step. Our in vivo data show, in neonatal plasma, at birth, detected C11-oxy C(21) steroids were 2-fold higher than C11-oxy C(19) steroids, ~755.6 and ~393.1 ng/dL, respectively. Conversely, in PCOS patients, the C11-oxy C(19) steroids were present at higher levels compared to the C11-oxy C(21) steroids and also compared to controls, showing a greater shunt towards the production of C(19) and C11-oxy C(19) steroids. In benign prostatic hyperplasia patients, suffering from a relatively undefined condition in terms of adrenal steroids, the C11-oxy C(19) steroid levels were 8-fold higher in circulation compared to the C11-oxy C(21) steroids, with prominent C11-oxy steroids quantified as 11KPROG (190.5 ng/dL), 11keto-dihydroprogesterone (278.9 ng/dL), 11OHA4 (2598.8 ng/dL), 11keto-androstenedione (415.4 ng/dL) and 11KDHT (183.7 ng/dL). We propose that the 11OHA4- and C11-oxy C(21) backdoor pathways may identify novel steroid biomarkers in clinically defined and undefined conditions. Indeed, accurate quantification of these steroid hormones has been facilitated by UPC(2)-MS/MS with comprehensive steroid profile analysis potentially aiding neonatal screening, identifying rare enzymatic deficiencies, determining hyperandrogenism in males and females −promising routinely applicable superiority over other analytical methods, with this study underscoring this promise.