Jiangxi Maternal & Child Health Hospital

BackgroundPolycystic ovary syndrome (PCOS) is an endocrine disorders and characterized by polycystic ovary morphol. and oligomenorrhea, affecting fertility and health condition of female around the world.The causative factors of PCOS are complex, and genetic structure remains a long-standing medical challenge in genetics.Previous genome-wide association study (GWAS) showed that Wing-less-related integration site (Wnt) signaling is the most affected pathway among PCOS-related risk genes, and genetic mutations in the Wnt/β-catenin signaling may lead to abnormal development of PCOS.ObjectiveTo explore the possibility of axis inhibitor-2 (AXIN2) variants in Chinese women with PCOS and assess their pathogenicities.MethodsA total of 365 Chinese women with PCOS and 905 women without PCOS as control were recruited from Jiangxi Provincial Maternal and Child Health Hospital, All of the 11 exons and flanking regions of the AXIN2 gene were amplified by polymerase chain reaction (PCR), the potential variants were analyzed by Sanger sequencing.The evolutionary conservation anal. of the identified Axin-2 mutant was analyzed among 15 vertebrates from human to zebrafish.The protein structure change was analyzed between the wild-type and mutation-type.The pathogenicity of AXIN2 variant was further analyzed in silico.ResultsWe totally identified 7 genetic variants of AXIN2 in this study, including 4 synonymous and 3 missense.Among them, we find a rare deleterious missense variant [p.R714W (c.2140C > T)].The allele frequencies of this variant were 0.82% and 0.17% in PCOS cases and matched controls, resp.And it was ranging from 7.89e-5 to 1.47e-4 in public databases.Fisher′s exact test indicated that the allele frequencies in PCOS were p < 0.05 compared to both the controls and the databases.Especially, the mutant amino acid site is highly conserved in vertebrates, while the mutation changed the 714th arginine to tryptophan resulting in significant change in the protein structural of Axin-2.ConclusionIn this study, we identified a rare deleterious missense AXIN2 mutation [p.R714W (c.2140C > T)] in Chinese women with PCOS, and this mutant is probably pathogenic.This study may provide a new perspective on revealing the genetic variation of PCOS.

To determine the variation of serum AMH levels in healthy Chinese women and establish AMH reference ranges accordingly.

This prospective cross-sectional multicenter study was designed to enroll healthy Chinese women of reproductive age (20-39 years) and perimenopausal age (40-49 years) from five reproductive centers in different regions of China. The study began in May 2022 and finished in February 2023. Age-specific 2.5th-97.5th percentiles AMH reference ranges were established. Multivariable linear regressions were undertaken to analyze the association of serum AMH with different demographic and clinical variables, including antral follicle count (AFC).

1113 healthy Chinese women were enrolled, including 614 of premenopausal age and others of reproductive age. The AMH (ng/ml) reference ranges for Chinese women of reproductive age were 0.87-9.89 (20-24 years), 0.42-8.24 (25-29 years), 0.34-7.46 (30-34 years), and 0.28-5.66 (35-39 years). For perimenopausal women, their reference ranges were 0.12-4.63 (40-41 years), 0.01-4.12 (42-43 years), 0.01-2.65 (44-45 years), 0.01-1.90 (46-47 years), and 0.01-1.08 (48-49 years). The regression of AMH on AFC adjusted by age is Log10(AMH)=0.2594-0.0235*Age + 0.0632*AFC.

This study established the age-specific serum AMH reference ranges for healthy Chinese women of reproductive and premenopausal age, and observed that the consistent decrease of AMH after 20 years accelerated around the beginning of perimenopause (40 years).

The global prevalence of premature ovarian failure (POF) is from 3.1 % to 4.3 %, which is a multifactorial disease including genetics, environmental and medical factors. However, the occurrence of POF is not well understood. To further explore the potential mechanism of POF, cyclophosphamide (CTX) was used to construct the model of POF. Additionally, the occurrence of POF was related to oxidative stress, apoptosis, proliferation and others. Ca²⁺ is essential for almost all life processes. Therefore, TRPM2-deficient mice was used to explore the role of Ca²⁺ in POF. The results indicated that the mouse serum E₂ and AMH levels decreased, FSH and LH levels increased, and the activity of antioxidant enzymes including CAT, SOD and GSH decreased with mtROS accumulation in the ovary, thereby causing ovarian DNA damage, promoting ovarian cell apoptosis and inhibiting cell proliferation after wild-type mice exposed to CTX. Notably, these indicators have improved after TRPM2^-/-. Based on these, we have further proved that the activation TRPM2 channel could lead to intracellular Ca²⁺ overload, plentiful Ca²⁺ bind to calmodulin accompanied with mitochondrial ROS accumulation, thereby activating AMPK/p53 signaling pathway, inducing proliferation arrest and excessive apoptosis. We hope to provide therapeutic targets to prevent the occurrence of POF by studying the potential mechanism of POF.