Granulosa cell-derived miR-379-5p regulates macrophage polarization in polycystic ovarian syndrome

Polycystic ovarian syndrome (PCOS) is associated with hyperandrogenemia and ovarian antral follicle growth arrest. We have previously demonstrated that androgen-induced exosomal release of miR-379-5p (miR379) from preantral follicle granulosa cells increases the proliferation of target cells via phosphoinositide-dependent kinase 1 (PDK1) upregulation. Androgen also increases inflammatory M1 macrophage abundance, but reduces anti-inflammatory M2 polarization in rat antral and preovulatory follicles. However, the role of small extracellular vesicles (sEVs; also known as exosomes) secretion in determining the cellular content and function of miRNAs in exosome-receiving cells is largely unknown. Our objectives were to determine: 1) the regulatory role of granulosa cells (GC)-derived exosomal miR379 on macrophage polarization and ovarian inflammation; 2) whether miR379-induced M1 polarization regulates GC proliferation; and 3) if this regulated process is follicular stage-specific. Compared with non-PCOS subjects, PCOS subjects had a higher M1/M2 ratio, supporting the concept that PCOS is an inflammatory condition. Ovarian overexpression of miR379 increased the number of M1 macrophages and the M1/M2 ratio in preantral follicles specifically. Transfection of macrophages with a miR379 mimic reduced the cellular content of PDK1 and induced M0 -> M1 polarization; whereas its inhibitor polarized M0 -> M2. Conditioned media from macrophages transfected with miR379 mimic and follicular fluid from PCOS subjects had higher galectin-3 content, a pro-inflammatory cytokine which specifically suppresses human antral follicle GC proliferation. These results indicate that miR379 inhibits M2 macrophage polarization, a condition which suppresses GC proliferation in a follicle stage-dependent manner, as exhibited in PCOS.

Automated summary

Polycystic ovarian syndrome (PCOS) is associated with increased levels of androgen and arrested follicle growth. Previous studies have shown that androgen-induced release of miR-379-5p from follicle granulosa cells promotes cell proliferation through PDK1 upregulation. Androgen also affects macrophage polarization and increases the inflammatory M1 phenotype in rat follicles. However, the role of small extracellular vesicles (exosomes) in mediating the effects of miRNAs on receiving cells is not well understood. This study aimed to investigate the regulatory role of exosomal miR-379 from granulosa cells on macrophage polarization and ovarian inflammation, and its impact on granulosa cell proliferation in a stage-specific manner.