Altered cargo proteins of human plasma endothelial cell–derived exosomes in atherosclerotic cerebrovascular disease

Abstract

Plasma endothelial cell-derived exosomes (EDEs) and platelet-derived exosomes (PDEs) were precipitated and enriched separately by immunospecific absorption procedures for analyses of cargo proteins relevant to atherosclerosis. EDEs had usual exosome size and marker protein content, and significantly higher levels than PDEs of the endothelial proteins vascular cell adhesion molecule-1 (VCAM-1) and endothelial nitric oxide synthase, whereas PDEs had significantly higher levels of platelet glycoprotein VI. EDE levels of VCAM-1, von Willebrand factor, platelet-derived growth factor (PDGF)-BB, angiopoietin-1, and lysyl oxidase-2 and the cerebrovascularselective proteins glucose transporter 1, permeability-glycoprotein, and large neutral amino acid transporter 1 were significantly higher for 18 patients with cerebrovascular disease (CeVD) than for 18 age-and gender-matched control subjects. PDE levels of PDGF-AA, platelet glycoprotein VI, integrin-linked kinase-1, high mobility group box-1 protein, chemokine CXCL4, and thrombospondin-1 were significantly higher in patients with CeVD than in control subjects, but differences were less with greater overlaps than for EDE proteins. EDE levels of Yes-associated protein (YAP) were higher and of P(S127)-YAP lower in patients with CeVD than in control subjects, consistent with heightened activity of this mechanical force-sensitive system in atherosclerosis. Elevated EDE and PDE levels of atherosclerosis-promoting proteins in CeVD justify clinical studies of their potential value as biomarkers.-Goetzl, E. J., Schwartz, J. B., Mustapic, M., Lobach, I. V., Daneman, R., Abner, E. L., Jicha, G. A. Altered cargo proteins of human plasma endothelial cell-derived exosomes in atherosclerotic cerebrovascular disease. FASEB J. 31, 3689-3694 (2017). www.fasebj.org

Exosomal biomarkers in Down syndrome and Alzheimer's disease

“…As discussed in this review, toxic Aβ peptides and toxic species of P-Tau, neuroinflammation, and oxidative stress can all alter exosome content, secretion rate, and the cell-to-cell messages they carry. Several reports indicate that exosomes produced under pathological conditions can either confer protection or increased pathology to their target cells (Borland and Vilhardt 2017; Chiarini et al 2017; Goetzl et al 2016; Goetzl et al 2017; Kapogiannis et al 2015; Kouwaki et al 2016; Lee et al 2016; Li et al 2013; Papandreou and Tavernarakis 2017). Further study of these processes will provide new insights into how AD pathology spreads throughout the brain and the role of exosomes in this process.…”

“…Because of this, the cargo carried by CNS-derived exosomes may serve as markers of underlying CNS changes that occur in advance of changes in circulating proteins. Because CNS-derived exosomes have unique surface markers that reflect their cell of origin, targeted examinations of neuron-, astrocyte-, or endothelial cell-derived exosomes can be employed (Fiandaca et al 2015; Kapogiannis et al 2015; Goetzl et al 2016; Goetzl et al 2017). After targeted selection using antibodies directed against the cell-type-specific surface marker(s) for an exosome population of interest, examination of specific exosomal cargos may be used to probe this unique biological niche that may provide a “liquid biopsy snapshot” of current cellular processes.…”

Extracellular vesicles in cardiovascular homeostasis and disease

“…Elevated glucose to diabetic levels induces an increase in both the rate of EV release and the size of EVs produced by endothelial cells in culture [5]. Circulating endothelial-derived EVs in patients with cerebrovascular atherosclerosis exhibit altered protein expression consistent with known changes in endothelial cell phenotype [6], and vascular smooth muscle cells (VSMCs) exposed to atherogenic lipoproteins exhibit altered EV microRNA profiles [7**]. These changes in EV size and/or cargo could alter communication between cells to respond to an extracellular cue and may exacerbate pathological conditions by disrupting homeostatic paracrine signaling.…”

Endothelial-derived exosomes demonstrate a link between endothelial innate inflammation and brain dysfunction and injury in aging

“…All brain MRIs were rated for burden of WMH by a board-certified neurologist (FME). At the time of enrollment in the current study platelet-poor-plasma was prepared according to published methods 25,26 . Plasma samples were aliquoted and stored at -80°C.…”

“…Platelet-poor plasma was prepared from 6 ml of venous blood and stored in 0.5 ml aliquots at -80°C as previously described 26 and EDE were enriched as per previously published protocol 26 . Briefly, after depletion of platelets, EDE exosomes were enriched by sequential immunoprecipitation with two biotinylated monoclonal antibodies to CD31 (MEM-05, Thermo Fisher Scientific) and then CD146 (Novus Biologicals, Littleton, CO, USA) prior to lysis of exosomes for quantification of cargo proteins via ELISA.…”