{"paper_id":"ac01e698-829e-4d6c-986f-a14728b76695","body_text":"Abstract\nEndometriosis is a multifactorial gynecological disease, with angiogenesis as a key hallmark. The role of exosomal microRNAs (miRNAs) in endometriosis is not well understood. This study investigates differentially expressed exosomal miRNAs linked to angiogenesis in endometriosis, clarifies their molecular mechanisms, and identifies potential targets. Primary endometrial stromal cells (ESCs) were cultured, and exosomes were extracted. In a co-culture system, ESC-derived exosomes were taken up by human umbilical vein endothelial cells (HUVECs). Endometriosis implant-ESC-derived exosomes (EI-EXOs) significantly promoted HUVEC proliferation, migration and tube formation compared to normal endometrium-exosomes (NE-EXOs), a finding consistent in vivo in mice. MiRNA sequencing and bioinformatics identified differentially expressed miR-21-5p from EI-EXOs, confirmed by RT-qPCR. The miR-21-5p inhibitor or GW4869 attenuated EI-EXO-induced HUVEC proliferation, migration, and tube formation. TIMP3 overexpression diminished the pro-angiogenic effect of EI-EXOs, which was reversed by adding EI-EXOs or upregulating miR-21-5p. These findings validate the crosstalk between ESCs and HUVECs mediated by exosomal miR-21-5p, and confirm the miR-21-5p-TIMP3 axis in promoting angiogenesis in endometriosis.\nKey messages\n-\nESC-derived exosomes were found to be taken up by recipient cells, i.e. HUVECs. Functionally, endometriosis implant-ESC-derived exosomes (EI-EXOs) could significantly promote the proliferation, migration and tube formation of HUVECs compared to normal endometrium-exosomes (NE-EXOs).\n-\nThrough miRNA sequencing and bioinformatics analysis, differentially expressed miR-21-5p released by EI-EXOs was chosen, as confirmed by qRT-PCR.\n-\nmiR-21-5p inhibitor or GW4869 was found to attenuate the proliferation, migration, and tube formation of HUVECs induced by EI-EXOs. In turn, TIMP3 overexpression diminished the pro-angiogenic effect of EI-EXOs, and this angiogenic phenotype was reversed once EI-EXOs were added or miR-21-5p was upregulated.\nSimilar content being viewed by others\nData availability\nData sharing is not applicable to this article as no datasets were generated or analysed during the current study.\nReferences\nAllaire C, Bedaiwy MA, Yong PJ (2023) Diagnosis and management of endometriosis. 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Am J Obstet Gynecol 223(4):557 .e1-557.e11\nFunding\nThis work was supported by the National Natural Science Foundation of China (Grant No. 81971359), National Natural Science Foundation of Heilongjiang Province (Grant No. LH2019H027), and Key R&D project of Heilongjiang Province (Grant No. GA21C008) to G.Z., and National Natural Science Foundation of China (Grant No. 82101725) to Y.C.\nAuthor information\nAuthors and Affiliations\nContributions\nL.S. and G.Z. designed this study. L.S., Y.C., J.W., D.W., L.Y., X.W., Y.L., and J.G. conducted the experiments, analysed the data, and performed statistical analysis. L.S. wrote the manuscript with help from Y. C., J. W., G. Z. L. Y., X. W., and Y. L. provided scientific support and discussion. G.Z. critically reviewed the manuscript and provided scientific support for experiments. All authors revised and commented on the manuscript drafts.\nCorresponding author\nEthics declarations\nEthics statement\nThe studies involving human specimens were reviewed and approved by the Ethical Committee of the First Affiliated Hospital of Harbin Medical University, and written informed consent was obtained from all participants. The animal study protocol was reviewed and approved by the Committee on the Ethical Use of Animals of the Harbin Medical University.\nConflict of interest\nAll the authors declare no competing interests.\nAdditional information\nPublisher’s note\nSpringer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.\nSupplementary Information\nBelow is the link to the electronic supplementary material.\nRights and permissions\nSpringer 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.\nAbout this article\nCite this article\nSun, L., Cheng, Y., Wang, J. et al. Exosomal miR-21-5p derived from endometrial stromal cells promotes angiogenesis by targeting TIMP3 in ovarian endometrial cysts. J Mol Med 102, 1327–1342 (2024). https://doi.org/10.1007/s00109-024-02483-z\nReceived:\nRevised:\nAccepted:\nPublished:\nVersion of record:\nIssue date:\nDOI: https://doi.org/10.1007/s00109-024-02483-z","source_license":"public-domain-us","license_restricted":false}