{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,14]],"date-time":"2026-07-14T23:50:31Z","timestamp":1784073031021,"version":"3.55.0"},"reference-count":32,"publisher":"MDPI AG","issue":"7","license":[{"start":{"date-parts":[[2024,6,28]],"date-time":"2024-06-28T00:00:00Z","timestamp":1719532800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"St. George Hospital Cancer Research Trust Fund","award":["190481"],"award-info":[{"award-number":["190481"]}]},{"name":"St. George Hospital Cancer Research Trust Fund","award":["RG220652"],"award-info":[{"award-number":["RG220652"]}]},{"name":"St. George and Sutherland Medical Research Foundation","award":["190481"],"award-info":[{"award-number":["190481"]}]},{"name":"St. George and Sutherland Medical Research Foundation","award":["RG220652"],"award-info":[{"award-number":["RG220652"]}]},{"name":"Prostate and Breast Cancer Foundation (PBCF)","award":["190481"],"award-info":[{"award-number":["190481"]}]},{"name":"Prostate and Breast Cancer Foundation (PBCF)","award":["RG220652"],"award-info":[{"award-number":["RG220652"]}]},{"name":"University International Postgraduate Award (UIPA) Scholarship at UNSW Sydney, Australia","award":["190481"],"award-info":[{"award-number":["190481"]}]},{"name":"University International Postgraduate Award (UIPA) Scholarship at UNSW Sydney, Australia","award":["RG220652"],"award-info":[{"award-number":["RG220652"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Metabolites"],"abstract":"<jats:p>Conventional diagnostic tools for prostate cancer (PCa), such as prostate-specific antigen (PSA), transrectal ultrasound (TRUS), digital rectal examination (DRE), and tissue biopsy face, limitations in individual risk stratification due to invasiveness or reliability issues. Liquid biopsy is a less invasive and more accurate alternative. Metabolomic analysis of extracellular vesicles (EVs) holds a promise for detecting non-genetic alterations and biomarkers in PCa diagnosis and risk assessment. The current research gap in PCa lies in the lack of accurate biomarkers for early diagnosis and real-time monitoring of cancer progression or metastasis. Establishing a suitable approach for observing dynamic EV metabolic alterations that often occur earlier than being detectable by other omics technologies makes metabolomics valuable for early diagnosis and monitoring of PCa. Using four distinct metabolite extraction approaches, the metabolite cargo of PC3-derived large extracellular vesicles (lEVs) was evaluated using a combination of methanol, cell shearing using microbeads, and size exclusion filtration, as well as two fractionation chemistries (pHILIC and C18 chromatography) that are also examined. The unfiltered methanol\u2013microbeads approach (MB-UF), followed by pHILIC LC-MS\/MS for EV metabolite extraction and analysis, is effective. Identified metabolites such as L-glutamic acid, pyruvic acid, lactic acid, and methylmalonic acid have important links to PCa and are discussed. Our study, for the first time, has comprehensively evaluated the extraction and separation methods with a view to downstream sample integrity across omics platforms, and it presents an optimised protocol for EV metabolomics in PCa biomarker discovery.<\/jats:p>","DOI":"10.3390\/metabo14070367","type":"journal-article","created":{"date-parts":[[2024,7,1]],"date-time":"2024-07-01T09:06:07Z","timestamp":1719824767000},"page":"367","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":9,"title":["Optimising Extracellular Vesicle Metabolomic Methodology for Prostate Cancer Biomarker Discovery"],"prefix":"10.3390","volume":"14","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-9823-079X","authenticated-orcid":false,"given":"Mahmoud Assem","family":"Hamed","sequence":"first","affiliation":[{"name":"St. George and Sutherland Clinical Campuses, School of Clinical Medicine, University of New South Wales (UNSW) Sydney, Kensington, NSW 2052, Australia"},{"name":"Cancer Care Centre, St. George Hospital, Kogarah, NSW 2217, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5338-0869","authenticated-orcid":false,"given":"Valerie","family":"Wasinger","sequence":"additional","affiliation":[{"name":"Bioanalytical Mass Spectrometry Facility, Mark Wainwright Analytical Centre, University of New South Wales (UNSW) Sydney, Kensington, NSW 2052, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Qi","family":"Wang","sequence":"additional","affiliation":[{"name":"St. George and Sutherland Clinical Campuses, School of Clinical Medicine, University of New South Wales (UNSW) Sydney, Kensington, NSW 2052, Australia"},{"name":"Cancer Care Centre, St. George Hospital, Kogarah, NSW 2217, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Joanna","family":"Biazik","sequence":"additional","affiliation":[{"name":"Electron Microscope Unit, Mark Wainwright Analytical Centre, University of New South Wales (UNSW) Sydney, Kensington, NSW 2052, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4957-4895","authenticated-orcid":false,"given":"Peter","family":"Graham","sequence":"additional","affiliation":[{"name":"St. George and Sutherland Clinical Campuses, School of Clinical Medicine, University of New South Wales (UNSW) Sydney, Kensington, NSW 2052, Australia"},{"name":"Cancer Care Centre, St. George Hospital, Kogarah, NSW 2217, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"David","family":"Malouf","sequence":"additional","affiliation":[{"name":"Department of Urology, St. George Hospital, Kogarah, NSW 2217, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Joseph","family":"Bucci","sequence":"additional","affiliation":[{"name":"St. George and Sutherland Clinical Campuses, School of Clinical Medicine, University of New South Wales (UNSW) Sydney, Kensington, NSW 2052, Australia"},{"name":"Cancer Care Centre, St. George Hospital, Kogarah, NSW 2217, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8910-9787","authenticated-orcid":false,"given":"Yong","family":"Li","sequence":"additional","affiliation":[{"name":"St. George and Sutherland Clinical Campuses, School of Clinical Medicine, University of New South Wales (UNSW) Sydney, Kensington, NSW 2052, Australia"},{"name":"Cancer Care Centre, St. George Hospital, Kogarah, NSW 2217, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2024,6,28]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"5225","DOI":"10.2147\/CMAR.S250829","article-title":"Biomarkers That Differentiate Benign Prostatic Hyperplasia from Prostate Cancer: A Literature Review","volume":"12","author":"McNally","year":"2020","journal-title":"Cancer Manag. 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