title: Cellular stress conditions are reflected in the protein and RNA content of endothelial cell-derived exosomes.
abstract: BACKGROUND
  The healthy vascular endothelium, which forms the barrier between blood and the surrounding tissues, is known to efficiently respond to stress signals like hypoxia and inflammation by adaptation of cellular physiology and the secretion of (soluble) growth factors and cytokines. Exosomes are potent mediators of intercellular communication. Their content consists of RNA and proteins from the cell of origin, and thus depends on the condition of these cells at the time of exosome biogenesis. It has been suggested that exosomes protect their target cells from cellular stress through the transfer of RNA and proteins. We hypothesized that endothelium-derived exosomes are involved in the endothelial response to cellular stress, and that exosome RNA and protein content reflect the effects of cellular stress induced by hypoxia, inflammation or hyperglycemia.
  
  
  METHODS
  We exposed cultured endothelial cells to different types of cellular stress (hypoxia, TNF-α-induced activation, high glucose and mannose concentrations) and compared mRNA and protein content of exosomes produced by these cells by microarray analysis and a quantitative proteomics approach.
  
  
  RESULTS
  We identified 1,354 proteins and 1,992 mRNAs in endothelial cell-derived exosomes. Several proteins and mRNAs showed altered abundances after exposure of their producing cells to cellular stress, which were confirmed by immunoblot or qPCR analysis.
  
  
  CONCLUSIONS
  Our data show that hypoxia and endothelial activation are reflected in RNA and protein exosome composition, and that exposure to high sugar concentrations alters exosome protein composition only to a minor extend, and does not affect exosome RNA composition.
authors.affiliation: Department of Nephrology and Hypertension, UMC Utrecht, Utrecht, The Netherlands.
authors.forename: Olivier G
authors.initials: OG
authors.lastname: de Jong
authors.forename: Marianne C
authors.initials: MC
authors.lastname: Verhaar
authors.forename: Yong
authors.initials: Y
authors.lastname: Chen
authors.forename: Pieter
authors.initials: P
authors.lastname: Vader
authors.forename: Hendrik
authors.initials: H
authors.lastname: Gremmels
authors.forename: George
authors.initials: G
authors.lastname: Posthuma
authors.forename: Raymond M
authors.initials: RM
authors.lastname: Schiffelers
authors.forename: Marjan
authors.initials: M
authors.lastname: Gucek
authors.forename: Bas W M
authors.initials: BW
authors.lastname: van Balkom
keywords: RNA; exosomes; extracellular vesicles; hypoxia; proteomics; tumor necrosis factor alpha