The Role of Extracellular Vesicles in the Pathophysiology of Endometriosis

Abstract

Endometriosis (EM) is a chronic inflammatory condition characterized by the growth of endometrial-like tissue outside of the uterine cavity. The precise molecular mechanisms driving lesion formation and subsequent development remains unknown. Recently, there has been growing interest in extracellular vesicles (EVs) as regulators of pivotal processes such as proliferation, vascularization, and inflammation, which are integral to EM development. In this study, we unveil a novel role played by EVs in EM pathophysiology. We isolated EVs from plasma, ectopic lesions, and eutopic endometrium of EM patients and healthy controls, and thoroughly investigated EM-derived EVs using nanoparticle tracking analysis, transmission electron microscopy, multiplex bead-based flow cytometry assay, small RNA sequencing, proteomics, and lipidomics. Preliminary results from these ongoing experiments indicate that EVs from EM patients exhibit distinctive surface markers reflecting their cellular origins. Furthermore, next-generation sequencing of miRNA profiles in EVs derived from EM patient tissues revealed unique miRNA signatures in EVs from ectopic lesions, showing promise as potential biomarkers. Additionally, mass spectrophotometry-based proteomic analysis of EVs from patient plasma and ectopic and eutopic samples unveiled stage-specific proteomic profiles. Functional investigations performed in endometriotic epithelial and endothelial cell lines using EVs from patient plasma and control samples clearly indicate roles in autocrine uptake and paracrine cell proliferation, implicating their involvement in endometriosis. Multiplex cytokine analysis of cell supernatants obtained from epithelial and stromal cells in response to patient and control plasma derived EVs unveiled robust signatures of inflammatory cytokines known to contribute to disease progression. Collectively, these findings suggest that EVs associated with endometriosis harbor distinct cargo and contribute to the disease's pathophysiology by influencing inflammation, angiogenesis, and proliferation within the microenvironment of endometriotic lesions.