Journal of Inflammation Research (May 2023)
Single-Cell Profiling Uncovers the Roles of Endometrial Fibrosis and Microenvironmental Changes in Adenomyosis
Abstract
Weipin Niu,1,2,* Yinuo Zhang,3,* Hongyun Liu,4 Na Liang,5 Li Xu,6 Yalin Li,7 Wei Yao,8 Wei Shi,8 Zhiyong Liu1 1Central Laboratory, The Affiliated Hospital of Shandong University of Traditional Chinese Medicine, Jinan, People’s Republic of China; 2Innovative Institute of Chinese Medicine and Pharmacy, Shandong University of Traditional Chinese Medicine, Jinan, People’s Republic of China; 3College of Traditional Chinese Medicine, Shandong University of Traditional Chinese Medicine, Jinan, People’s Republic of China; 4Department of Gynecology, Linyi Central Hospital, Linyi, People’s Republic of China; 5Department of Traditional Chinese Medicine, The First Affiliated Hospital of Shandong First Medical University, Shandong Provincial Qianfoshan Hospital, Jinan, People’s Republic of China; 6Department of Gynecology and Obstetrics, The Second Affiliated Hospital of Shandong University of Traditional Chinese Medicine, Jinan, People’s Republic of China; 7First College of Clinical Medicine, Shandong University of Traditional Chinese Medicine, Jinan, People’s Republic of China; 8Department of Gynecology, The Affiliated Hospital of Shandong University of Traditional Chinese Medicine, Jinan, People’s Republic of China*These authors contributed equally to this workCorrespondence: Zhiyong Liu, Central Laboratory, The Affiliated Hospital of Shandong University of Traditional Chinese Medicine, Jinan, Shandong, 250014, People’s Republic of China, Email [email protected] Wei Shi, Department of Gynecology, The Affiliated Hospital of Shandong University of Traditional Chinese Medicine, No. 16369 Jingshi Road, Jinan, Shandong, 250014, People’s Republic of China, Email [email protected]: Adenomyosis (AM) is a common benign uterine disorder that has deleterious effects on women’s health. However, the pathogenesis of AM is not clearly understood. We aimed to investigate the pathophysiological changes and molecular mechanism in AM.Methods: Single-cell RNA sequencing (scRNA-seq) was employed to construct a transcriptomic atlas of various cell subsets from the ectopic endometrium (EC) and eutopic endometrium (EM) of one AM patient and evaluate differential expression. The Cell Ranger software pipeline (version 4.0.0) was applied to conduct sample demultiplexing, barcode processing and mapping reads to the reference genome (human GRCh38). Different cell types were classified with markers with the “FindAllMarkers” function, and differential gene expression analysis was performed with Seurat software in R. The findings were confirmed by Reverse Transcription Real-Time PCR using samples from three AM patients.Results: We identified nine cell types: endothelial cells, epithelial cells, myoepithelial cells, smooth muscle cells, fibroblasts, lymphocytes, mast cells, macrophages and unknown cells. A number of differentially expressed genes, including CLO4A1, MMP1, TPM2 and CXCL8, were identified from all cell types. Functional enrichment showed that aberrant gene expression in fibroblasts and immune cells was related to fibrosis-associated terms, such as extracellular matrix dysregulation, focal adhesion and the PI3K-Akt signaling pathway. We also identified fibroblast subtypes and determined a potential developmental trajectory related to AM. In addition, we identified increased cell–cell communication patterns in EC, highlighting the imbalanced microenvironment in AM progression.Conclusion: Our results support the theory of endometrial–myometrial interface disruption for AM, and repeated tissue injury and repair could lead to increased fibrosis in the endometrium. Therefore, the present study reveals the association between fibrosis, the microenvironment, and AM pathogenesis. This study provides insight into the molecular mechanisms regulating AM progression.Keywords: AM, scRNA-seq, ectopic endometrium, eutopic endometrium, extracellular matrix, fibrosis
