囊泡 circLDLR 耗竭解除 miR-1294 功能抑制并通过 CYP19A1 抑制多囊卵巢综合征卵泡液中雌二醇的产生。
Depletion of exosomal circLDLR in follicle fluid derepresses miR-1294 function and inhibits estradiol production via CYP19A1 in polycystic ovary syndrome.
机构信息
Department of Assisted Reproduction, and Clinical and Translational Research Center, Shanghai First Maternity and Infant Hospital, Tongji University School of Medicine, Shanghai, China.
出版信息
Aging (Albany NY). 2020 Jul 10;12(15):15414-15435. doi: 10.18632/aging.103602.
Polycystic ovary syndrome (PCOS) is a common endocrine and metabolic disorder in reproductive women and is characterized by polycystic ovaries, hyperandrogenism and chronic anovulation. Abnormal folliculogenesis is considered as a common characteristic of PCOS. Our aim is to identify the altered circRNA expression profile in exosomes isolated from follicular fluid (FF) of PCOS patients to investigate the molecular function of exosomal circRNA, as a vital mediator in follicular microenvironment, in the aetiology and pathobiology of PCOS. In this study, the circRNA expression profile of FF exosomes were compared between PCOS and control patients by RNA sequencing (N=5 vs 5). Sixteen circRNAs showed significantly different expression. GO and KEGG pathway analyses indicated that their parental genes were enriched in PCOS-related pathways, including ovarian steroidogenesis, aldosterone synthesis and secretion, and Jak-STAT signaling. Among sixteen differentially expressed circRNAs, hsa_circ_0006877 (circLDLR) was processed from its parental LDLR (low density lipoprotein receptor) transcript, which participated in ovarian steroidogenesis. Its depletion in PCOS FF exosomes was further verified in an additional cohort (N=25 vs 25) by qRT-PCR. And a circLDLR-miR-1294-CYP19A1 competing endogenous RNA (ceRNA) network was predicted by cytoscape software, and confirmed by luciferase assay and correlative expression in the cumulus cells of PCOS patients. Mechanistically, the intercellular transfer of functional circLDLR assay and its withdrawal experiments in KGN cells showed that depleting circLDLR in exosomes increased miR-1294 expression and inhibited CYP19A1 expression in recipient cells, as well as reduced their estrogen (E2) secretion. Our findings revealed a ceRNA network of circLDLR and provided new information on abnormal follicle development in PCOS.
多囊卵巢综合征(PCOS)是一种常见的生殖期女性内分泌和代谢紊乱疾病,其特征为多囊卵巢、高雄激素血症和慢性无排卵。异常的卵泡发生被认为是 PCOS 的共同特征。我们的目的是鉴定来源于 PCOS 患者卵泡液(FF)的外泌体中改变的 circRNA 表达谱,以研究外泌体 circRNA 作为卵泡微环境中的重要介质在 PCOS 的发病机制和病理生物学中的分子功能。在这项研究中,通过 RNA 测序(N=5 对 5)比较了 PCOS 患者和对照组患者 FF 外泌体的 circRNA 表达谱。有 16 个 circRNA 表现出显著差异表达。GO 和 KEGG 通路分析表明,它们的亲本基因富集在与 PCOS 相关的通路中,包括卵巢甾体生成、醛固酮合成和分泌以及 Jak-STAT 信号通路。在 16 个差异表达的 circRNA 中,hsa_circ_0006877(circLDLR)来源于其亲本 LDLR(低密度脂蛋白受体)转录本,该 circRNA 参与卵巢甾体生成。在另外一个包含 25 对患者的队列中,通过 qRT-PCR 进一步验证了 PCOS FF 外泌体中 circLDLR 的缺失。并通过 Cytoscape 软件预测了 circLDLR-miR-1294-CYP19A1 竞争内源 RNA(ceRNA)网络,并通过荧光素酶测定和 PCOS 患者卵丘细胞的相关性表达进行了验证。从机制上讲,功能性 circLDLR 的细胞间转移及其在 KGN 细胞中的撤出实验表明,外泌体中 circLDLR 的耗竭增加了受体细胞中 miR-1294 的表达并抑制了 CYP19A1 的表达,同时减少了其雌激素(E2)的分泌。我们的研究结果揭示了 circLDLR 的 ceRNA 网络,并为 PCOS 中异常卵泡发育提供了新的信息。