SOX9: a novel mediator of the ovulatory process in the human ovary

The Eunice Kennedy Shriver National Institute of Child Health and Human Development (NICHD), under the Child Health and Human Development Extramural Research program (CFDA 93.865), awarded $419,014 to the University of Kentucky Research Foundation beginning September 1, 2025, and concluding August 31, 2027. This project grant funds investigator-initiated biomedical research designed to advance understanding of reproductive health and female fertility outcomes. The research will be conducted at the awardee's facility in Lexington, Kentucky.

The funded project investigates SOX9 (Sex-Determining Region Y (SRY)-Box Transcription Factor 9) as a critical mediator of the ovulatory process in the human ovary. Research activities will examine how SOX9 regulation contributes to ovulation and corpus luteum development in women, addressing a significant gap in understanding ovarian disorders that account for approximately 30 percent of infertility cases. The research deliverables include scientific findings regarding SOX9's role as a downstream-regulated gene of progesterone receptor and its function in controlling gene expression essential for successful ovulation and luteinization, with results expected to provide a foundation for improved fertility management and treatment approaches.

Abstract

Ovarian disorders in women, including defects in ovulation and subsequent corpus luteum (CL) development, account for up to 30% of all infertility cases. Understanding the mechanism underlying these processes is critically important, as this knowledge provides a foundation for our ability to aid or manage female fertility. The mid-cycle gonadotropin surge triggers dramatic changes in the dominant follicle, culminating in ovulation and transformation into the CL. One essential step elicited by the gonadotropin surge is the induction of specific transcription factors that directly control the transcription of genes crucial for successful ovulation and luteinization. However, our knowledge of regulatory actions of LH/HCG-induced ovulatory transcription factors remains limited, especially in humans. Our preliminary data shed light on SOX9 (SEX-determining region y (SRY)-box transcription factor 9) as a critical transcriptional regulator involved in periovulatory processes. IT has long been accepted that SOX9 is expressed at a low level or repressed in the developing ovary. This active suppression of SOX9 in granulosa cells is crucial for maintaining ovarian phenotype. Challenging this belief, our preliminary data show that SOX9 expression is markedly up-regulated in granulosa cells of dominant follicles obtained from normally cycling women after the LH surge. Furthermore, our pilot study shows that SOX9 is a direct downstream-regulated gene of progesterone/progesterone receptor (PGR), an essential mediator of the ovulatory process and regulates the expression of star and CYP11A1 and progesterone production in human granulosa/lutein cells. Based on these data, we hypothesize that SOX9 is a human-specific ovulatory gene that plays a critical role in ovulation and luteinization by regulating the expression of genes involved in the periovulatory process. This hypothesis will be tested utilizing in vivo ovarian tissue samples obtained from normally cycling women during various ovulatory phases and a primary human granulosa/lutein cell culture model that can recapitulate in vivo changes in ovulatory gene expression and hormone profiles. In aim 1, we will determine the expression pattern of SOX9 and its downstream genes in follicular cells isolated from dominant follicles and whole dominant follicles obtained from normally cycling women throughout the periovulatory period. In aim 2, we will identify SOX9-regulated downstream genes and assess the impact of SOX9 knockdown on granulosa/lutein cell function. The proposed is in line with R21 mechanisms, as IT will not only, for the first time, demonstrate the ovulatory up-regulation of SOX9 in human ovulatory follicles but also enhance our understanding of how the LH surge/HCG - P4/PGR - SOX9 pathway controls the human-specific ovulatory process and luteinization in the ovary. This fundamental knowledge could be directly applied to uncovering the molecular mechanisms underlying anovulatory disorders or luteal insufficiency, as well as developing strategies to improve the management of female fertility.