Ovarian reserve status in young women is associated with altered gene expression in membrana granulosa cells

Abstract

Diminished ovarian reserve (DOR) is a challenging diagnosis of infertility, as there are currently no tests to predict who may become affected with this condition, or at what age. We designed the present study to compare the gene expression profile of membrana granulosa cells from young women affected with DOR with those from egg donors of similar age and to determine if distinct genetic patterns could be identified to provide insight into the etiology of DOR. Young women with DOR were identified based on FSH level in conjunction with poor follicular development during an IVF cycle (n = 13). Egg donors with normal ovarian reserve (NOR) comprised the control group (n = 13). Granulosa cells were collected following retrieval, RNA was extracted and microarray analysis was conducted to evaluate genetic differences between the groups. Confirmatory studies were undertaken with quantitative RT-PCR (qRT-PCR). Multiple significant differences in gene expression were observed between the DOR patients and egg donors. Two genes linked with ovarian function, anti-Mullerian hormone (AMH) and luteinizing hormone receptor (LHCGR), were further analyzed with qRT-PCR in all patients. The average expression of AMH was significantly higher in egg donors (adjusted P-value = 0.01), and the average expression of LHCGR was significantly higher in DOR patients (adjusted P-value = 0.005). Expression levels for four additional genes, progesterone receptor membrane component 2 (PGRMC2), prostaglandin E receptor 3 (subtype EP3) (PTGER3), steroidogenic acute regulatory protein (StAR), and StAR-related lipid transfer domain containing 4 (StarD4), were validated in a group consisting of five NOR and five DOR patients. We conclude that gene expression analysis has substantial potential to determine which young women may be affected with DOR. More importantly, our analysis suggests that DOR patients fall into two distinct subgroups based on gene expression profiles, indicating that different mechanisms may be involved during development of this pathology.

Aging-related premature luteinization of granulosa cells is avoided by early oocyte retrieval

“…That premature luteinization negatively impacts oocyte quality, fertilization and implantation is supported by Skiadas et al (2012) who demonstrated an association between low functional ovarian reserve in older women and premature luteinization, clinically characterized by higher peripheral LH and lower AMH levels. Also, oocyte numbers and top quality embryo numbers have been reported to be significantly higher in normal patients than in women with premature luteinization (Bosch et al 2003, Elnashar 2010).…”

Identification of the key regulating genes of diminished ovarian reserve (DOR) by network and gene ontology analysis

“…Although infertility treatments have been improved through assisted reproductive technology (ART) over the past 30 years, the molecular reasons of infertility have not been fully identified [31]. Women who respond well to fertility treatments are defined as having normal ovarian reserve (NOR).…”

Genetic associations with diminished ovarian reserve: a systematic review of the literature

“…These receptors are important in ovarian steroidogenesis, luteinization, and therefore, progesterone synthesis and fertility. Skiadas et al [42] found a 2.19-fold increased expression of the LHCGR gene in DOR patients over NOR patients. The increased gene expression of LHCGR in DOR patients could be explained by premature luteinization and upregulation of luteinizing hormone receptors on follicular cells, which might occur with DOR patients given the increased FSH in the follicular phase.…”

“…Typically, serum AMH values below 0.8 ng/mL have been found to be consistent with elevated FSH levels and severely diminished ovarian reserve [6]. Skiadas et al [42] examined membrana granulosa cells from women with DOR.…”