Evaluating the Effect of Exosome-Encapsulated miR-4289 on Menstrual Blood-Derived Mesenchymal Stem Cells from Endometriosis Patients

Endometriosis is a benign yet chronic gynecological disorder characterized by dysregulation of processes such as inflammation, angiogenesis, migration, apoptosis, and proliferation. Menstrual blood-derived endometrial stem cells play a crucial role in the retrograde development and progression of endometriotic lesions. To evaluate the therapeutic potential of exosomes derived from menstrual blood-derived stem cells, exosomes from non-endometriotic MenSCs (NE-MenSCs), both unmodified (Exo) and transfected with miR-4289, were applied as treatments to MenSCs from the endometriosis cell line (E-MenSCs). Publicly available databases were used to identify key genes and signaling pathways implicated in endometriosis, from which miR-4289 was selected as an effective regulatory microRNA. Following treatment, cellular migration was assessed by scratch assays; gene expression was evaluated via real-time PCR; protein levels of ROS, IL-10, and IL-1β were measured by ELISA; and ESR1, CTNNB1, and Ki67 levels were determined by Western blotting. The results indicate that treatments significantly reduced the expression of genes associated with inflammation, proliferation, migration, and the Wnt/β-catenin pathway. Scratch assays and reductions in MMP9 expression suggest decreased migration in the Exo and miR-Exo groups. The expression of CTNNB1, IL-1β, and IL-10 was significantly downregulated in treated groups compared to E-MenSCs. In addition, KRAS and IDO1 expression levels were significantly decreased following treatment, and Ki67 protein levels were notably reduced in the miR and miR-Exo groups. These findings highlight the therapeutic potential of MenSC-derived exosomes loaded with miR-4289 as a promising and novel strategy for treating endometriosis. Graphical Abstract Similar content being viewed by others Data Availability The datasets used and/or analyzed during the current study are available in supplementary section. Abbreviations - MenSCs: - Menstrual Stem Cells - NE-MenSCs: - Non-Endometriosis Menstrual Stem Cells - E-MenSCs: - Endometriosis Menstrual Stem Cells - Exo group: - Exosome Group - miR-Exo group: - microRNA-Exosome Group - miR group: - microRNA Group - BB: - Backbone - EV: - Extracellular vesicle - DLS: - Dynamic light scattering - CM: - Conditioned medium - TEM: - Transmission electron microscopy

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Not applicable. Funding Not applicable. Author information Authors and Affiliations Contributions SM, ASh performed the lab work. SM, ASh reviewed the relevant literature and wrote the manuscript. NH, EE drafted and revised the manuscript. SM, ASh made the figures and tables. All authors reviewed and approved the manuscript **.**. Corresponding author Ethics declarations Competing Interest The authors declare no competing interests. Ethical Approval and Consent to Participate This experimental investigation conducted in the field of endometriosis, received ethical approval from the Islamic Azad University Ethics Committee (ethical code: IR.IAU.SRB.REC.1401.312). NE-MenSCs and E-MenSCs cell lines were procured from the Department of Cell Biology and Regenerative Medicine of ACECR, Qom Branch. Characterization of these cell lines was confirmed by flow cytometry, as reported in a published article (Asl et al. 2023). We then investigated treatments using exosomes and microRNA. Consent for Publication Not applicable. Additional information Publisher’s Note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations. Supplementary Information Below is the link to the electronic supplementary material. Rights and permissions Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law. About this article Cite this article Mahmoudi, S., Sheikholeslami, A., Roodbari, N.H. et al. Evaluating the Effect of Exosome-Encapsulated miR-4289 on Menstrual Blood-Derived Mesenchymal Stem Cells from Endometriosis Patients. Biochem Genet (2025). https://doi.org/10.1007/s10528-025-11265-2 Received: Accepted: Published: Version of record: DOI: https://doi.org/10.1007/s10528-025-11265-2