Luo Man, Tian Xiaona, Li Li, Zhang Guomei, Liu Wenzhi, Mei Linlin, Li Haoran, You Xiaoyan, Zhang Dongmei, Zhou Mengsi, Xiao Cheng, Ye Jianping, Yang Xiaofeng
Department of Obstetrics and Gynecology, Zhengzhou Central Hospital Affiliated to Zhengzhou University, Zhengzhou, China.
Zhengzhou Key Laboratory of Endocrine Metabolism and Immunity in Polycystic Ovary Syndrome, Zhengzhou Central Hospital Affiliated to Zhengzhou University, Zhengzhou, China.
Front Cell Dev Biol. 2025 Aug 4;13:1633254. doi: 10.3389/fcell.2025.1633254. eCollection 2025.
Polycystic ovary syndrome (PCOS) is a common endocrine disorder characterized by elevated androgen levels and impaired follicular development. A hallmark of PCOS is the excessive proliferation of thecal cells (TCs), which are critical for androgen production. However, the molecular mechanisms underlying this aberrant cellular expansion remain incompletely understood.
A DHEA-induced mouse model was used to recapitulate the hormonal and ovarian features of human PCOS. Spatial transcriptomics was employed to profile gene expression in ovarian tissue at cellular resolution. Differential expression analysis, pathway enrichment, and spatial co-localization were performed to identify regulatory networks. Functional assays were conducted in cultured TCs using siRNA-mediated knockdown of target genes, and cell proliferation and cell cycle progression were evaluated using EdU incorporation and flow cytometry.
Spatial transcriptomic profiling revealed widespread transcriptional changes in the ovaries of PCOS mice, including a marked expansion of a TCs subpopulation with high Lrp2 expression. This subset exhibited enhanced activity in genes involved in androgen synthesis and cell cycle regulation. A signaling axis comprising Inhba, Smad2, and E2f4 was identified as a key regulator of this proliferative response, with all three genes co-expressed in the affected regions. Knockdown of any component of this axis significantly suppressed TCs proliferation , with the greatest effect observed upon Inhba silencing.
The Inhba/Smad2/E2f4 axis contributes to thecal cell hyperplasia and androgen excess in PCOS, and may serve as a mechanistic entry point for further investigation into the regulation of TCs proliferation in this disorder.
多囊卵巢综合征（PCOS）是一种常见的内分泌紊乱疾病，其特征为雄激素水平升高和卵泡发育受损。PCOS的一个标志是卵泡膜细胞（TCs）过度增殖，而卵泡膜细胞对雄激素的产生至关重要。然而，这种异常细胞扩张背后的分子机制仍未完全清楚。
使用脱氢表雄酮（DHEA）诱导的小鼠模型来模拟人类PCOS的激素和卵巢特征。采用空间转录组学技术在细胞分辨率下分析卵巢组织中的基因表达。进行差异表达分析、通路富集分析和空间共定位分析以确定调控网络。在培养的TCs中使用小干扰RNA（siRNA）介导的靶基因敲低进行功能测定，并使用EdU掺入法和流式细胞术评估细胞增殖和细胞周期进程。
空间转录组分析揭示了PCOS小鼠卵巢中广泛的转录变化，包括具有高Lrp2表达的TCs亚群的显著扩张。该亚群在参与雄激素合成和细胞周期调控的基因中表现出增强的活性。一条由抑制素βA（Inhba）、Smad2和E2f4组成的信号轴被确定为这种增殖反应的关键调节因子，这三个基因在受影响区域共表达。敲低该信号轴的任何一个成分均显著抑制TCs增殖，其中Inhba沉默时效果最为明显。
Inhba/Smad2/E2f4信号轴导致PCOS中卵泡膜细胞增生和雄激素过多，可能作为进一步研究该疾病中TCs增殖调控机制的切入点。
