Proteomic analysis of plasma and extracellular vesicles from subjects with impaired vascular health

Abstract

Abstract Cardiovascular diseases are the leading cause of mortality worldwide, with atherosclerosis and formation of arterial plaques being a major underlying cause. Rupture or erosion of the plaque fibrous cap can result in thrombus formation, arterial occlusion, and a stroke or myocardial infarction. Plaque changes, and endothelial cell barrier leakiness, may result in material leakage, including proteins and fragments into plasma either directly or in extracellular vesicles (EVs). Here, we report comparative LC-MS/MS analyses of plasma-derived EVs and plasma from subjects with impaired vascular status and healthy controls. Analysis of plasma-derived EVs detected 7228 peptides and 763 proteins, with 87 proteins being differentially abundant with these including arterial-cell species. Sub-group analysis based on biological sex showed no statistically significant differences for males, whereas females exhibited eight differentially expressed proteins. Subject age effects were minimal. Plasma analysis detected 4366 peptides and 497 proteins, with 188 proteins being significantly altered in abundance between the groups. Subgroup analysis by biological sex revealed 103 differentially expressed proteins in females and 84 in males. No differences were detected in specific collagen fragments. Gene Set Enrichment Analysis revealed altered biological processes related to immune regulation, humoral immune response, proteolysis, and cellular components including plasma lipoprotein particle, extracellular space, and membrane-associated structures. KEGG pathway analysis emphasized enrichment of pathways linked to complement and coagulation cascades, platelet activation, focal adhesion, endocytosis, and inflammation. Together, these data illustrate the potential of LC-MS/MS to examine the role of inflammation and arterial wall cells in shaping the proteome of EVs and plasma in health and disease.