Brennan Kieran, Vaiciuleviciute Raminta, Uzieliene Ilona, Pachaleva Jolita, Kasilovskiene Zaneta, Piesiniene Lina, Bernotiene Eiva, Mc Gee Margaret M
School of Biomolecular and Biomedical Science, University College Dublin, Belfield, Dublin 4, Ireland.
Conway Institute of Biomolecular and Biomedical Research, University College Dublin, Belfield, Dublin 4, Ireland.
Sci Rep. 2025 Apr 8;15(1):11974. doi: 10.1038/s41598-025-95818-w.
Several biomolecules have been previously associated with unexplained infertility (uIF) in blood and uterine samples, immune cells and their secreted factors, endometrial tissue, menstrual blood, serum, and stromal cells, however they do not comprehensively represent different uIF endotypes and their isolation/detection involves invasive diagnostic methods and lacks precision. This ex-vivo study was performed on extracellular vesicles (EVs) from menstrual blood collected on cycle day 2 from 9 fertile volunteers and 8 women with uIF. Menstrual blood serum (MBS) EVs were isolated from fertile and uIF MBS using Iodixanol Density Gradient Centrifugation and quantified by flow cytometry. EVs were characterized according to MISEV2023 guidelines. Comprehensive proteomic analysis of MBS EVs and EV-depleted MBS showed significant changes in uIF proteome, mostly affecting cell adhesion, immune response, apoptosis, response to oxidative stress and lipid metabolism. These processes were previously linked to pathologies of the female reproductive system but never investigated in uIF and were used to stratify patients into distinct molecular endotypes. Area under the curve (AUC) analysis was used to determine the optimum set of biomarkers for each of the uIF molecular endotypes. These findings provide new insights into uIF that could facilitate personalised treatment approaches.
先前已有多种生物分子与血液和子宫样本、免疫细胞及其分泌因子、子宫内膜组织、月经血、血清和基质细胞中的不明原因不孕症（uIF）相关，然而它们并未全面代表不同的uIF内型，且其分离/检测涉及侵入性诊断方法且缺乏精准性。本体外研究对来自9名生育能力正常的志愿者和8名uIF女性在月经周期第2天采集的月经血中的细胞外囊泡（EV）进行了研究。使用碘克沙醇密度梯度离心法从生育能力正常者和uIF者的月经血血清（MBS）中分离出EV，并通过流式细胞术进行定量。根据MISEV2023指南对EV进行了表征。对MBS EV和去除EV的MBS进行的综合蛋白质组学分析显示，uIF蛋白质组有显著变化，主要影响细胞黏附、免疫反应、细胞凋亡、氧化应激反应和脂质代谢。这些过程此前与女性生殖系统疾病有关，但从未在uIF中进行过研究，并且被用于将患者分层为不同的分子内型。使用曲线下面积（AUC）分析来确定每种uIF分子内型的最佳生物标志物组合。这些发现为uIF提供了新的见解，可能有助于个性化治疗方法的制定。
