Henarejos-Castillo Ismael, Sanz Francisco José, Solana-Manrique Cristina, Sebastian-Leon Patricia, Medina Ignacio, Remohi José, Paricio Nuria, Diaz-Gimeno Patricia
IVI-RMA Global Research Alliance, IVI Foundation, Instituto de Investigación Sanitaria La Fe (IIS La Fe), Av. Fernando Abril Martorell 106, Valencia, 46026, Spain.
Department of Pediatrics, Obstetrics and Gynaecology, University of Valencia, Av. Blasco Ibáñez 15, Valencia, 46010, Spain.
Reprod Biol Endocrinol. 2024 Dec 4;22(1):153. doi: 10.1186/s12958-024-01325-4.
Ovarian failure (OF) is a multifactorial, complex disease presented by up to 1% of women under 40 years of age. Despite 90% of patients being diagnosed with idiopathic OF, the underlying molecular mechanisms remain unknown, making it difficult to personalize treatments for these patients in the clinical setting. Studying the presence and/or accumulation of SNVs at the gene/pathway levels will help describe novel genes and characterize disrupted biological pathways linked with ovarian failure.
Ad-hoc case-control SNV screening conducted from 2020 to 2023 of 150 VCF files WES data included Spanish IVF patients with (n = 118) and without (n = 32) OF (< 40 years of age; mean BMI 22.78) along with GnomAD (n = 38,947) and IGSR (n = 1,271; 258 European female VCF) data for pseudo-control female populations. SNVs were prioritized according to their predicted deleteriousness, frequency in genomic databases, and proportional differences across populations. A burden test was performed to reveal genes with a higher presence of SNVs in the OF cohort in comparison to control and pseudo-control groups. Systematic in-silico analyses were performed to assess the potential disruptions caused by the mutated genes in relevant biological pathways. Finally, genes with orthologues in Drosophila melanogaster were considered to experimentally validate the potential impediments to ovarian function and reproductive potential.
Eighteen genes had a higher presence of SNVs in the OF population (FDR < 0.05). AK2, CDC27, CFTR, CTBP2, KMT2C, and MTCH2 were associated with OF for the first time and their silenced/knockout forms reduced fertility in Drosophila. We also predicted the disruption of 29 sub-pathways across four signalling pathways (FDR < 0.05). These sub-pathways included the metaphase to anaphase transition during oocyte meiosis, inflammatory processes related to necroptosis, DNA repair mismatch systems and the MAPK signalling cascade.
This study sheds light on the underlying molecular mechanisms of OF, providing novel associations for six genes and OF-related infertility, setting a foundation for further biomarker development, and improving precision medicine in infertility.
卵巢功能衰竭（OF）是一种多因素的复杂疾病，在40岁以下的女性中发病率高达1%。尽管90%的患者被诊断为特发性OF，但其潜在的分子机制仍不清楚，这使得在临床环境中难以对这些患者进行个性化治疗。在基因/通路水平研究单核苷酸变异（SNV）的存在和/或积累，将有助于发现新基因，并确定与卵巢功能衰竭相关的生物通路中断情况。
2020年至2023年对150个VCF文件的全外显子组测序（WES）数据进行了针对性的病例对照SNV筛查，数据包括西班牙体外受精（IVF）患者（年龄<40岁；平均体重指数22.78），其中有OF的患者（n = 118）和无OF的患者（n = 32），以及用于虚拟对照女性群体的基因组聚集数据库（GnomAD，n = 38,947）和国际基因组样本资源（IGSR，n = 1,271；258个欧洲女性VCF）数据。根据SNV的预测有害性、在基因组数据库中的频率以及不同人群之间的比例差异对其进行优先级排序。进行负担检验以揭示与对照组和虚拟对照组相比，OF队列中SNV存在较多的基因。进行系统的计算机模拟分析，以评估突变基因在相关生物通路中造成的潜在破坏。最后，考虑在黑腹果蝇中有直系同源物的基因，以实验验证对卵巢功能和生殖潜力的潜在阻碍。
18个基因在OF人群中存在较多的SNV（错误发现率<0.05）。AK2、CDC27、CFTR、CTBP2、KMT2C和MTCH2首次与OF相关，它们的沉默/敲除形式降低了果蝇的生育力。我们还预测了四个信号通路中29个亚通路的破坏（错误发现率<0.05）。这些亚通路包括卵母细胞减数分裂过程中从中期到后期的转变、与坏死性凋亡相关的炎症过程、DNA错配修复系统和丝裂原活化蛋白激酶（MAPK）信号级联反应。
本研究揭示了OF的潜在分子机制，为六个基因与OF相关的不孕症提供了新的关联，为进一步开发生物标志物奠定了基础，并改善了不孕症的精准医学。
