Abstract

ABSTRACT Little is known about the composition and functional differences between extracellular vesicle (EV) subsets, such as microvesicles (MVs) and exosomes (EXOs), nor to what extent their cargo reflects the phenotypic state of the cell of origin. Brain endothelial cells are the constitutive part of the blood–brain barrier (BBB), a selective barrier that maintains brain homeostasis. BBB impairment is associated with several neuroinflammatory diseases with the pro‐inflammatory cytokine tumour necrosis factor (TNF) often playing a key role. In the present study, shotgun proteomics and parallel reaction monitoring (PRM)‐based targeted mass spectrometry were used to characterise brain endothelial cell‐released EVs, and to study how TNF exposure modulated EV protein cargoes. MVs were found to be enriched in mitochondrial and cytoskeletal proteins, whereas EXOs were enriched in adhesion, histone and ribosomal proteins. After stimulation with TNF, several proteins involved in TNF and NF‐κB signalling pathways, that were found to be differentially expressed in cells, were also differentially expressed in both MVs and EXOs. Thus, our results revealed some novel proteins as potentially useful candidates for discriminating between MVs and EXOs, together with additional evidence that cells “package” proteins in EVs systematically and according to their phenotypic state.

Keywords

MicrovesiclesCell biologyProteomicsBiologyExtracellular vesicleTumor necrosis factor alphaShotgun proteomicsStable isotope labeling by amino acids in cell cultureNeuroinflammationCytokinemicroRNAInflammationImmunologyBiochemistryGene

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Year
2017
Type
article
Volume
6
Issue
1
Pages
1302705-1302705
Citations
150
Access
Closed

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Vito Dozio, Jean‐Charles Sanchez (2017). Characterisation of extracellular vesicle‐subsets derived from brain endothelial cells and analysis of their protein cargo modulation after TNF exposure. Journal of Extracellular Vesicles , 6 (1) , 1302705-1302705. https://doi.org/10.1080/20013078.2017.1302705

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DOI
10.1080/20013078.2017.1302705