Endocrine Abstracts

Introduction: Altered intrauterine endocrine environments can ‘programme’ adverse health outcomes. Linkage between altered androgen exposure in utero and adverse offspring health is robust. For example, increased maternal androgen concentrations and PCOS in female offspring and dyslipidaemia in male offspring. We hypothesised that the liver was a major target for androgenic programming in utero and hepatic dysfunction would be present in offspring. We therefore examined the hepatic transcriptome and proteome in a non-biased approach, to understand if key hepatic functions are altered by prenatal androgens, and identify circulating biomarkers in adolescence, indicative of prenatal androgenic excess.

Methods: Ovine fetuses were directly injected with 200 μl testosterone propionate (PA; 20 mg) or vehicle control (C), under ultrasound guidance at day 62 and 82 of gestation. Male fetuses were studied at day 90 of pregnancy (C, n=11; PA, n=6) and adolescents at 6 months postnatal age (C, n=14; PA, n=14). Hepatic transcriptome and proteome were determined using Illumina RNA sequencing and liquid chromatography-mass spectrometer (LC-MS/MS). Plasma proteins and analytes were measured using LC-MS/MS, ELISA or benchtop biochemistry autoanalysers. Statistical analysis between C and PA groups was performed using pairwise comparisons, with false discovery rate correction, accepting P<0.05 as significant.

Results: In fetal livers of PA males there was downregulated expression of transcripts involved in bile acid, bilirubin and xenobiotics export (ABCC2) (P<0.05). This was maintained in the transcriptome data from livers of adolescent PA males (P<0.05). However, in adolescence there was also altered expression of genes/proteins involved in liver damage and fibrosis. Data predicted increased oxidative stress and altered reactive oxygen species handling with downregulated phase II detoxification (GSS, GSTM1, GSTM4, GSTO1) (all P<0.05) and decreased protection against reactive oxygen species (GPX4, GSR, PRDX5, XDH) (all P<0.05). This was associated with increased expression of genes associated with liver fibrosis (COL4A1, COL4A5, COL4A6, COL18A1, COL27A1, FGF7, FGFR2, FGFR3, SMAD3, SMAD7, TGFBI) (all P<0.05). Plasma analysis of adolescent males revealed decreased levels of antioxidant enzymes (CAT, SOD3, GPX3) (all P<0.05) and increased circulating bilirubin and collagens (COL1A1, COL1A2, COL5A1) (all P<0.05).

Conclusions: This is the first study documenting that direct fetal male androgen overexposure results in decreased hepatic detoxification capacity, with potential for liver damage and fibrosis. In addition to mechanistic understanding of the journey from in utero androgenic overexposure to adolescent hepatic health issues, we observed the echoes of hepatic alterations reflected in the circulation, providing utility for biomarker development.