Evaluating the effect of exosome-encapsulated miR-4289 on menstrual blood-derived mesenchymal stem cells from endometriosis patients

Evaluating the effect of exosome-encapsulated miR-4289 on menstrual blood-derived mesenchymal stem cells from endometriosis patients | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Evaluating the effect of exosome-encapsulated miR-4289 on menstrual blood-derived mesenchymal stem cells from endometriosis patients Solmaz Mahmoudi, Nasim Hayati Roodbari, Ehsan Ehsani, Azar Sheikholeslami This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-6558623/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 11 Nov, 2025 Read the published version in Biochemical Genetics → Version 1 posted 11 You are reading this latest preprint version Abstract Endometriosis is a benign, yet chronic gynecological disorder characterized by deregulation in processes such as inflammation, angiogenesis, migration, apoptosis, and proliferation. Menstrual bloodderived 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 bloodderived stem cells, exosomes from non-endometriotic MenSCs (NE-MenSCs), both unmodified (Exo) and transfected with miR-4289, were applied as treatments to MenSCs from endometriosis patients (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/βcatenin pathway. Scratch assays and reductions in MMP9 expression suggest decreased migration in the Exo and miRExo 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 reduced notably in the miR and miRExo groups. These findings provide preliminary evidence for the therapeutic potential of MenSCderived exosomes, particularly when loaded with miR-4289, as a novel treatment approach for endometriosis. Menstrual blood Mesenchymal stem cells microRNA Exosome Transfection Endometriosis Full Text Additional Declarations No competing interests reported. Supplementary Files PCRfile.xlsx Elisa.rar Graphpad.rar Flowcytometry.docx WesternBlotting.rar scratch.rar Cite Share Download PDF Status: Published Journal Publication published 11 Nov, 2025 Read the published version in Biochemical Genetics → Version 1 posted Editorial decision: Revision requested 27 May, 2025 Reviews received at journal 27 May, 2025 Reviews received at journal 16 May, 2025 Reviewers agreed at journal 13 May, 2025 Reviewers agreed at journal 07 May, 2025 Reviewers agreed at journal 07 May, 2025 Reviewers agreed at journal 07 May, 2025 Reviewers invited by journal 07 May, 2025 Editor assigned by journal 30 Apr, 2025 Submission checks completed at journal 30 Apr, 2025 First submitted to journal 29 Apr, 2025 You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-6558623","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":454355896,"identity":"beddd681-209d-401c-b5cb-eb7f0f7face6","order_by":0,"name":"Solmaz Mahmoudi","email":"","orcid":"","institution":"SR.C, Islamic Azad University","correspondingAuthor":false,"prefix":"","firstName":"Solmaz","middleName":"","lastName":"Mahmoudi","suffix":""},{"id":454355897,"identity":"b6de7e33-4a62-4169-ae1f-b7cfa3168e48","order_by":1,"name":"Nasim Hayati Roodbari","email":"","orcid":"","institution":"SR.C, Islamic Azad 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