Characterization and proteomic analysis of endometrial stromal cell-derived small extracellular vesicles.

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Extracellular vesicles (sEVs) have emerged as modulators of the disease microenvironment, thereby supporting disease progression. However, the potential role of EVs and their content to the pathophysiology of endometriosis remain unclear.To investigate whether the EVs from eutopic (Eu) and ectopic (Ec) endometrial stromal cells (ESCs) differ with respect to protein composition and role in endometriosis.Human eutopic and ectopic endometrium derived ESCs were isolated from samples of the same patients (n=3). sEVs were isolated from ESCs via ultracentrifugation; these sEVs were characterized by western blotting, transmission electron microscopy and nanoparticle tracking analysis and analyzed using mass spectrometry. The potential role of ectopic ESCs-derived sEVs (EcESCs-sEVs) in endometriosis was explored by assaying their effects on cell viability/proliferation, migration, and angiogenesis.In total, 105 ESCs-sEVs-associated proteins were identified from EcESCs-sEVs and EuESCs-sEVs by mass spectrometry analysis. The protein content differed between EcESCs-sEVs and EuESCs-sEVs, with Annexin A2 (ANXA2) being the most prominent difference-present in EcESCs-sEVs but not EuESCs-sEVs. We also found that sEVs-ANXA2 regulates the motility, proliferation, and angiogenesis of ESCs via the ERK/STAT3 pathway. Notably, treatment of ESCs with sEVs-ANXA2 resulted in increased proliferation and motility, suggesting that sEVs-ANXA2 may be involved in regulating endometriosis. Our data suggest that EcESCs-sEVs-ANXA2 regulates the motility and the angiogenic potential of ESCs, implying a role for sEVs-ANXA2 in the pathogenesis of endometriosis.The study of sEVs-ANXA2 from ectopic endometriotic cells uncovers a new mechanism of endometriosis progression and will inform the development of novel therapeutic strategies.

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