Shao Wenwen, Ju Hongmei, Xiahou Zhikai, Fang Sheng, Yan Rugen, Li Chunyan, Xu Yuan, Cai Pingping
College of First Clinical Medicine, Shandong University of Traditional Chinese Medicine, Jinan, China.
Department of Gynecology, Affiliated Hospital of Shandong University of Traditional Chinese Medicine, Jinan, China.
Front Immunol. 2025 Oct 13;16:1680849. doi: 10.3389/fimmu.2025.1680849. eCollection 2025.
Endometriosis (EM) is a chronic gynecological disorder that affects 5% to 10% of women of childbearing age, often causing pelvic pain and infertility. Fibrosis is a hallmark of EM progression, yet its underlying molecular drivers remain poorly understood. Emerging progress in single-cell and spatial transcriptomic technologies offer new opportunities to unravel the cellular heterogeneity and intercellular interactions driving fibrotic and immune remodeling in EM lesions.
We performed an integrative multi-omics analysis combining single-cell RNA sequencing (scRNA-seq) and spatial transcriptomics to dissect fibroblast heterogeneity and cell-cell communication networks in EM. ScRNA-seq data from 15 EM patients (GSE213216) were processed to identify transcriptionally distinct fibroblast subpopulations. Functional enrichment (GO, GSEA), stemness estimation (CytoTRACE), and trajectory inference were applied to reveal lineage plasticity. CellChat was used to infer intercellular signaling networks, and spatial transcriptomic data from two ectopic lesions (GSM6690475, GSM6690476) were analyzed to validate the spatial distribution of key ligand-receptor interactions.
We identified 35 cell clusters across EM lesions, with Fibroblast and T/NK cells as dominant populations. Fibroblast were divided into five subtypes, which were associated with extracellular matrix remodeling, immune interactions, and metabolic regulation. Notably, the C2 Fibroblast subpopulation exhibited high proliferative capacity and stemness characteristics, and mediated signaling pathways involved in immune and fibrotic responses through FN1. Spatial transcriptomic analysis confirmed the local enrichment of these Fibroblast in ectopic lesions, where they were associated with regions of active signaling.
This study revealed the transcriptional and spatial heterogeneity of Fibroblast in EM syndrome, and identified the C2 Fibroblast subpopulation as a may represent key driver of fibrosis and immune regulation. Our integrated omics approach provided new mechanistic insights into the pathogenesis of EM and pointed out new targets for therapeutic intervention.
子宫内膜异位症（EM）是一种慢性妇科疾病，影响5%至10%的育龄女性，常导致盆腔疼痛和不孕。纤维化是EM进展的一个标志，但其潜在的分子驱动因素仍知之甚少。单细胞和空间转录组技术的新进展为揭示驱动EM病变中纤维化和免疫重塑的细胞异质性和细胞间相互作用提供了新机会。
我们进行了一项整合多组学分析，结合单细胞RNA测序（scRNA-seq）和空间转录组学，以剖析EM中纤维母细胞的异质性和细胞间通信网络。对来自15名EM患者（GSE213216）的scRNA-seq数据进行处理，以识别转录上不同的纤维母细胞亚群。应用功能富集（GO、GSEA）、干性估计（CytoTRACE）和轨迹推断来揭示谱系可塑性。使用CellChat推断细胞间信号网络，并分析来自两个异位病变（GSM6690475、GSM6690476）的空间转录组数据，以验证关键配体-受体相互作用的空间分布。
我们在EM病变中鉴定出35个细胞簇，以纤维母细胞和T/NK细胞为主。纤维母细胞分为五个亚型，与细胞外基质重塑、免疫相互作用和代谢调节相关。值得注意的是，C2纤维母细胞亚群表现出高增殖能力和干性特征，并通过FN1介导参与免疫和纤维化反应的信号通路。空间转录组分析证实这些纤维母细胞在异位病变中局部富集，在那里它们与活跃信号区域相关。
本研究揭示了EM综合征中纤维母细胞的转录和空间异质性，并确定C2纤维母细胞亚群可能是纤维化和免疫调节的关键驱动因素。我们的整合组学方法为EM的发病机制提供了新的机制见解，并指出了治疗干预的新靶点。
