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Our approach focuses on extracellular vesicle (EV) biomarker discovery by directly profiling the proteome of EVs enriched from frozen biobanked endometrial tumours. We analysed nine tissue samples to compare three clinical subgroups\u2014low BMI (Body Mass Index) Endometrioid, high BMI Endometrioid, and Serous (any BMI)\u2014identifying proteins related to histological subtype, BMI, and shared secreted proteins. Using collagenase digestion and size exclusion chromatography, we successfully enriched generous quantities of EVs (range 204.8\u20131291.0\u00a0\u00b5g protein: 1.38\u00a0\u00d7\u00a010<jats:sup>11<\/jats:sup>\u20131.10\u00a0\u00d7\u00a010<jats:sup>12<\/jats:sup> particles), characterised by their size (\u223c150\u00a0nm), expression of EV markers (CD63\/81), and proposed endometrial cancer markers (L1CAM, ANXA2). Mass spectrometry\u2010based proteomic profiling identified 2075 proteins present in at least one of the 18 samples. Compared to cell lysates, EVs were successfully depleted for mitochondrial and blood proteins and enriched for common EV markers and large secreted proteins. Further analysis highlighted significant differences in EV protein profiles between the high BMI subgroup and others, underlining the impact of comorbidities on the EV secretome. Interestingly, proteins differentially abundant in tissue subgroups were largely not also differential in matched EVs. This research identified secreted proteins known to be involved in endometrial cancer pathophysiology and proposed novel diagnostic biomarkers (EIF6, MUC16, PROM1, SLC26A2).<\/jats:p>","DOI":"10.1002\/pmic.202300055","type":"journal-article","created":{"date-parts":[[2024,4,21]],"date-time":"2024-04-21T22:24:55Z","timestamp":1713738295000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":5,"title":["The tumour\u2010derived extracellular vesicle proteome varies by endometrial cancer histology and is confounded by an obesogenic environment"],"prefix":"10.1002","volume":"24","author":[{"given":"Anastasiia","family":"Artuyants","sequence":"first","affiliation":[{"name":"Department of Molecular Medicine and Pathology The University of Auckland  Auckland New Zealand"},{"name":"Auckland Cancer Society Research Centre The University of Auckland  Auckland New 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