COUP-TFII regulates early bipotential gonad signaling and commitment to ovarian progenitors

Abstract

AbstractThe absence of expression of the Y-chromosome linked testis-determining geneSRYin early supporting gonadal cells (ESGC) of bipotential gonads leads to ovarian development. However, genetic variants inNR2F2/COUP-TFII represent a novel cause ofSRY-negative 46,XX testicular/ovotesticular differences of sex development (T/OT-DSD). Thus, we hy-pothesized that COUP-TFII is part of the ovarian developmental network. We examinedNR2F2/COUP-TFII expression and the genetic network under its regulation in human gonadal cells by analyzing single cell RNA-sequencing datasets of fetal gonads, differentiating induced pluripotent stem cells into bipotential gonad-like cellsin vitro, and generating aNR2F2knockout (KO) in the human granulosa-like cell line COV434.NR2F2expression is highly upregulated during the bipotential gonad development, being detected in ESGCs. We identified thatNR2F2ablation in COV434 cells downregulated markers of ESGC and pre-granulosa cells, suggesting that COUP-TFII has a role in maintaining a multipotent state necessary for commitment to the ovarian development. We propose that impairment of COUP-TFII function may disrupt the transcriptional plasticity of ESGCs and instead drive them into commitment to the testicular pathway.