title: Proteomics analysis of vesicles isolated from plasma and urine of prostate cancer patients using a multiplex, aptamer-based protein array.
abstract: Proteomics analysis of biofluid-derived vesicles holds enormous potential for discovering non-invasive disease markers. Obtaining vesicles of sufficient quality and quantity for profiling studies has, however, been a major problem, as samples are often replete with co-isolated material that can interfere with the identification of genuine low abundance, vesicle components. Here, we used a combination of ultracentrifugation and size-exclusion chromatography to isolate and analyse vesicles of plasma or urine origin. We describe a sample-handling workflow that gives reproducible, quality vesicle isolations sufficient for subsequent protein profiling. Using a semi-quantitative aptamer-based protein array, we identified around 1,000 proteins, of which almost 400 were present at comparable quantities in plasma versus urine vesicles. Significant differences were, however, apparent with elements like HSP90, integrin αVβ5 and Contactin-1 more prevalent in urinary vesicles, while hepatocyte growth factor activator, prostate-specific antigen-antichymotrypsin complex and many others were more abundant in plasma vesicles. This was also applied to a small set of specimens collected from men with metastatic prostate cancer, highlighting several proteins with the potential to indicate treatment refractory disease. The study provides a practical platform for furthering protein profiling of vesicles in prostate cancer, and, hopefully, many other disease scenarios.
authors.affiliation: Division of Cancer and Genetics, School of Medicine, Cardiff University, Cardiff, United Kingdom.|Velindre Cancer Centre, Cardiff, United Kingdom.|Cardiff School of Health Sciences, Cardiff Metropolitan University, Cardiff, United Kingdom.
authors.forename: Joanne Louise
authors.initials: JL
authors.lastname: Welton
authors.affiliation: Division of Cancer and Genetics, School of Medicine, Cardiff University, Cardiff, United Kingdom.
authors.forename: Paul
authors.initials: P
authors.lastname: Brennan
authors.affiliation: Division of Cancer and Genetics, School of Medicine, Cardiff University, Cardiff, United Kingdom.|Velindre Cancer Centre, Cardiff, United Kingdom.
authors.forename: Mark
authors.initials: M
authors.lastname: Gurney
authors.affiliation: Division of Cancer and Genetics, School of Medicine, Cardiff University, Cardiff, United Kingdom.|Velindre Cancer Centre, Cardiff, United Kingdom.
authors.forename: Jason Paul
authors.initials: JP
authors.lastname: Webber
authors.affiliation: Division of Cancer and Genetics, School of Medicine, Cardiff University, Cardiff, United Kingdom.|Velindre Cancer Centre, Cardiff, United Kingdom.
authors.forename: Lisa Kate
authors.initials: LK
authors.lastname: Spary
authors.affiliation: Metabolomics Unit, CIC bioGUNE, CIBERehd, Bizkaia Technology Park, Derio, Spain.
authors.forename: David Gil
authors.initials: DG
authors.lastname: Carton
authors.affiliation: Metabolomics Unit, CIC bioGUNE, CIBERehd, Bizkaia Technology Park, Derio, Spain.
authors.forename: Juan Manuel
authors.initials: JM
authors.lastname: Falcón-Pérez
authors.affiliation: Department of Computer Science, College of Science, Swansea University, United Kingdom.
authors.forename: Sean Peter
authors.initials: SP
authors.lastname: Walton
authors.affiliation: Division of Cancer and Genetics, School of Medicine, Cardiff University, Cardiff, United Kingdom.|Velindre Cancer Centre, Cardiff, United Kingdom.
authors.forename: Malcolm David
authors.initials: MD
authors.lastname: Mason
authors.affiliation: Division of Cancer and Genetics, School of Medicine, Cardiff University, Cardiff, United Kingdom.|Velindre Cancer Centre, Cardiff, United Kingdom.
authors.forename: Zsuzsanna
authors.initials: Z
authors.lastname: Tabi
authors.affiliation: Division of Cancer and Genetics, School of Medicine, Cardiff University, Cardiff, United Kingdom.|Velindre Cancer Centre, Cardiff, United Kingdom; ClaytonA@cardiff.ac.uk.
authors.forename: Aled
authors.initials: A
authors.lastname: Clayton
keywords: plasma; protein array; proteomics; urine prostate