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Endometrial Mesenchymal Stem Cell-Derived Exosomal miR-4669 Promotes EMT in Adenomyosis by Inducing M2 Macrophage
Yingying Qiu1,2, Xinjun Wei1, Jian Cao3
1Department of Obstetrics and Gynecology, Affiliated Hospital of Integrated Traditional Chinese and Western Medicine, Nanjing University of Chinese Medicine, Nanjing, 210028, China.
Abstract:
Adenomyosis (AM), a gynecological disorder that severely affects female reproductive health. AM-associated macrophage (AAM) polarization-induced epithelial-mesenchymal transition (EMT) is a key driver of AM progression. In this study, we investigated the role and underlying mechanisms of endometrial mesenchymal stem cell (eMSC)-derived exosomes in regulating AAM polarization and the subsequent EMT of endometrial epithelial cells (EECs). In vitro coculture studies revealed that AM eutopic eMSCs markedly induced M2 macrophage polarization via exosomes and promoted EMT of EECs. Differentially expressed microRNAs (DE-miRNAs) between exosomes derived from normal eMSCs (N-eMSCs) and AM eutopic eMSCs (A-eMSCs) were identified using miRNA sequencing and miR-4669 was found to be the most significantly upregulated miRNA. Internalization of exosomal miR-4669 by macrophages induced their polarization toward the M2 phenotype and promoted the EMT of EECs. Mechanistic analysis using luciferase assay, mRNA sequencing, and rescue experiments revealed that miR-4669 induced M2 macrophage polarization via downregulation of DUSP6 and activation of MAPK/ERK signaling. The polarized M2 macrophages promoted the EMT of ISK cells via TGF-β1 secretion. In an AM xenograft mouse model, miR-4669 depletion inhibited AM progression by targeting the DUSP6/ERK1/2 pathway in macrophages. Overall, AM A-eMSC-derived exosomal miR-4669 facilitates M2 macrophage polarization by targeting the DUSP6/ERK signaling pathway, thereby promoting EMT of EECs via TGF-β1 secretion. These findings open avenues for developing novel preventive and therapeutic strategies for AM.
Insights
Endometrial mesenchymal stem cell exosomes carrying miR-4669 promote adenomyosis by polarizing macrophages and inducing epithelial-mesenchymal transition, offering new therapeutic targets.
Area of Science:
- Reproductive biology
- Cellular and molecular medicine
- Immunology
Background:
- Adenomyosis (AM) is a gynecological disorder impacting female reproductive health.
- Macrophage polarization and epithelial-mesenchymal transition (EMT) are key drivers of AM progression.
- Endometrial mesenchymal stem cells (eMSCs) and their exosomes play a role in AM pathogenesis.
Purpose of the Study:
- Investigate the role of eMSC-derived exosomes in regulating macrophage polarization and EMT in adenomyosis.
- Identify specific molecular mechanisms, including microRNAs, involved in this process.
- Explore therapeutic potential for adenomyosis based on these findings.
Main Methods:
- In vitro co-culture of eMSCs and macrophages with endometrial epithelial cells (EECs).
- miRNA sequencing to identify differentially expressed miRNAs in exosomes.
- Luciferase assays, mRNA sequencing, and rescue experiments to elucidate molecular pathways.
- In vivo studies using an adenomyosis xenograft mouse model.
Main Results:
- eMSC-derived exosomes from adenomyosis patients induced M2 macrophage polarization and promoted EMT in EECs.
- Exosomal miR-4669 was significantly upregulated in AM eMSCs and promoted M2 polarization and EMT.
- miR-4669 targeted DUSP6, activating the MAPK/ERK signaling pathway in macrophages.
- Polarized M2 macrophages secreted TGF-β1, further promoting EEC EMT.
- In vivo, miR-4669 depletion in exosomes inhibited AM progression.
Conclusions:
- Adenomyosis eMSC-derived exosomal miR-4669 drives M2 macrophage polarization via the DUSP6/ERK pathway.
- This process promotes EEC EMT through TGF-β1 secretion, contributing to adenomyosis progression.
- Targeting exosomal miR-4669 presents a potential therapeutic strategy for adenomyosis.
