Comparative and integrated analysis of plasma extracellular vesicle isolation methods in healthy volunteers and patients following myocardial infarction.

Autor: Paget D; Division of Cardiovascular Medicine, Radcliffe Department of Medicine University of Oxford Oxford UK.; Department of Pharmacology University of Oxford Oxford UK., Checa A; Unit of Integrative Metabolomics, Institute of Environmental Medicine Karolinska Institute Stockholm Sweden., Zöhrer B; Respiratory Medicine Unit, Department of Medicine Solna and Center for Molecular Medicine Karolinska Institutet Stockholm Sweden.; Department of Respiratory Medicine and Allergy Karolinska University Hospital Stockholm Sweden., Heilig R; Target Discovery Institute, Centre for Medicines Discovery, Nuffield Department of Medicine University of Oxford Oxford UK., Shanmuganathan M; Division of Cardiovascular Medicine, Radcliffe Department of Medicine University of Oxford Oxford UK.; Acute Vascular Imaging Centre, Radcliffe Department of Medicine University of Oxford UK., Dhaliwal R; Sir William Dunn School of Pathology University of Oxford Oxford UK., Johnson E; Sir William Dunn School of Pathology University of Oxford Oxford UK., Jørgensen MM; Department of Clinical Medicine Aalborg University Aalborg Denmark.; Department of Clinical Immunology Aalborg University Hospital Aalborg Denmark., Bæk R; Department of Clinical Medicine Aalborg University Aalborg Denmark., Wheelock CE; Unit of Integrative Metabolomics, Institute of Environmental Medicine Karolinska Institute Stockholm Sweden.; Department of Respiratory Medicine and Allergy Karolinska University Hospital Stockholm Sweden.; Gunma University Initiative for Advanced Research (GIAR) Gunma University Showa-machi Maebashi Gunma Japan., Channon KM; Division of Cardiovascular Medicine, Radcliffe Department of Medicine University of Oxford Oxford UK.; Acute Vascular Imaging Centre, Radcliffe Department of Medicine University of Oxford UK., Fischer R; Target Discovery Institute, Centre for Medicines Discovery, Nuffield Department of Medicine University of Oxford Oxford UK., Anthony DC; Department of Pharmacology University of Oxford Oxford UK., Choudhury RP; Division of Cardiovascular Medicine, Radcliffe Department of Medicine University of Oxford Oxford UK.; Acute Vascular Imaging Centre, Radcliffe Department of Medicine University of Oxford UK., Akbar N; Division of Cardiovascular Medicine, Radcliffe Department of Medicine University of Oxford Oxford UK.
Jazyk: angličtina
Zdroj: Journal of extracellular biology [J Extracell Biol] 2022 Nov 23; Vol. 1 (11), pp. e66. Date of Electronic Publication: 2022 Nov 23 (Print Publication: 2022).
DOI: 10.1002/jex2.66
Abstrakt: Plasma extracellular vesicle (EV) number and composition are altered following myocardial infarction (MI), but to properly understand the significance of these changes it is essential to appreciate how the different isolation methods affect EV characteristics, proteome and sphingolipidome. Here, we compared plasma EV isolated from platelet-poor plasma from four healthy donors and six MI patients at presentation and 1-month post-MI using ultracentrifugation (UC), polyethylene glycol precipitation, acoustic trapping, size-exclusion chromatography (SEC) and immunoaffinity capture. The isolated EV were evaluated by Nanoparticle Tracking Analysis (NTA), Western blot, transmission electron microscopy (TEM), an EV-protein array, untargeted proteomics (LC-MS/MS) and targeted sphingolipidomics (LC-MS/MS). The application of the five different plasma EV isolation methods in patients presenting with MI showed that the choice of plasma EV isolation method influenced the ability to distinguish elevations in plasma EV concentration following MI, enrichment of EV-cargo (EV-proteins and sphingolipidomics) and associations with the size of the infarct determined by cardiac magnetic resonance imaging 6 months post-MI. Despite the selection bias imposed by each method, a core of EV-associated proteins and lipids was detectable using all approaches. However, this study highlights how each isolation method comes with its own idiosyncrasies and makes the comparison of data acquired by different techniques in clinical studies problematic.
Competing Interests: The Author(s) declare that there is no conflict of interest.
(© 2022 The Authors. Journal of Extracellular Biology published by Wiley Periodicals, LLC on behalf of the International Society for Extracellular Vesicles.)
Databáze: MEDLINE
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