Integrated Single-Cell and Spatial Transcriptomics Reveal Androgen-Driven Disruptions in PCOS Ovarian Microenvironment

Polycystic Ovary Syndrome (PCOS) is a widespread reproductive disorder with hyperandrogenemia and impaired follicular development. Granulosa cells (GC) and thecal cells (TC) are crucial during follicular development, but their responses to androgen remain poorly understood at subpopulation levels. Using single-cell RNA sequencing and spatial transcriptomics, we analyzed GC and TC subpopulations in Dehydroepiandrosterone-induced PCOS-like mice to determine the ovarian microenvironment characters. We observed gene expression changes in steroidogenesis and cell-cell communications, with significant landscape shifts in subtypes. GC subsets showed an inflammatory GC5 expansion and loss of luteinized GC7, while TC subsets exhibited an increased TC2 population with enhanced lipid metabolism and androgen response. Intercellular signaling was heightened, particularly with amplified ligand-receptor interactions for Ptn-Ncl and Mdk-Ncl. Steroidogenesis was altered for PCOS's steroid hormone abnormalities. These findings revealed androgen impact in the cell subpopulations of follicular microenvironment and underscored the importance of Ptn-Ncl and Mdk-Ncl in PCOS-associated impaired follicular development. Ovaries were collected from Control mice and PCOS-like mice. PCOS-like mice were generated by subcutaneous administration of Dehydroepiandrosterone (DHEA) into 25-day-old female ICR mice. The DHEA was dissolved in sesame oil at 10% by volume in 95% ethanol, and was administrated in each mouse of model group at 6 mg/100 g of body weight in 0.1 mL once a day over a period of 20 days. The control mice were injected with vehicle in the same way. The mice were housed under a pathogen-free condition with a 12-hour light/dark cycle, free access to food and water.