MON-192 Gestational exposure to excess testosterone disrupts maternal steroid homeostasis and perturbs the steroid-lipid association in sheep

Abstract Disclosure: N. Saadat: None. R. Saeed: None. B. Pallas: None. A.K. Vyas: None. R.J. Auchus: None. V. Padmanabhan: None. Steroid hormones play a key role in pregnancy and in the growth and development of the fetus. Hyperandrogenism during pregnancy may disrupt this steroid homeostatic balance leading to unfavorable metabolic profiles, pregnancy complications and adverse programming for diseases later in life of the offspring. We hypothesized that gestational exposure of excess testosterone will not only disrupt maternal steroid homeostasis but also perturb the steroid-lipid relationship. Using a well-established translationally relevant sheep model, we investigated the impact of gestational testosterone (T) excess on maternal delta 4 and delta 5 steroid panels and steroid-lipidome relationship at day 127.1 ± 5.3 of pregnancy (term 147 days) in T-treated (T =12, 100mg T-propionate in corn oil, days 30 to 90 of gestation twice weekly intramuscularly) and control (C =5, vehicle only) Suffolk sheep. Steroids were measured using liquid chromatography-tandem mass spectrometry and lipids by shotgun lipidomics. Principal component analysis showed a clear separation of steroid profiles of T-treated and C groups. The main impact of gestational T excess was on the delta 5 pathway, reflected as significant reduction in 17-OH pregnenolone, androstenediol, allopregnanolone, and androsterone, and a reduced trend in dehydroepiandrosterone (DHEA, p=0.08) and 17-OH allopregnanolone (p=0.06) levels. A trend for reduced androstenedione (p-0.08) and increased corticosterone (p=0.06) and a large magnitude non-significant increase in 11 deoxycorticosterone was observed in delta 4 pathway. Gestational T excess disrupted the enriched pathways related to lipid biosynthesis and metabolism that prevail in controls but instead showed an enrichment in pathways related to cholesterol biosynthesis, circadian clock-relation, transcriptional regulation, and liver steatosis. Of relevance, disrupted steroid-lipid associations in gestational T-treated group have links to complex disease-related profiles that centers on lipid metabolism and transport, cholesterol and glycogen biosynthesis of relevance to metabolism-related diseases such as hyperlipidemia, gestational diabetes and hypertension. Further, only a few lipid species showed association with individual steroids indicative of loss of majority of the homeostatic steroid-lipid associations evidenced in controls. This study provides a novel comprehensive insight into the steroid-lipid metabolism homeostasis that prevail during normal pregnancy and importantly highlights the negative impact of gestational T excess in disrupting this balance in a precocial animal model of translational relevance. Our findings of disruptions in steroids and lipid interactive pathways impacted by gestational T excess may point to novel intervention strategies for reducing the impact of hyperandrogenism on maternal-fetal outcomes. Funding: R01 HD099096 Presentation: Monday, July 14, 2025