title: Circulating Plasma Extracellular Microvesicle MicroRNA Cargo and Endothelial Dysfunction in Children with Obstructive Sleep Apnea.
abstract: Obese children are at increased risk for developing obstructive sleep apnea (OSA), and both of these conditions are associated with an increased risk for endothelial dysfunction (ED) in children, an early risk factor for atherosclerosis and cardiovascular disease. Although weight loss and treatment of OSA by adenotonsillectomy improve endothelial function, not every obese child or child with OSA develops ED. Exosomes are circulating extracellular vesicles containing functional mRNA and microRNA (miRNA) that can be delivered to other cells, such as endothelial cells.
  
  
  
  To investigate whether circulating exosomal miRNAs of children with OSA differentiate based on endothelial functional status.
  
  
  
  Obese children (body mass index z score >1.65) and nonobese children were recruited and underwent polysomnographic testing (PSG), and fasting endothelial function measurements and blood draws in the morning after PSG. Plasma exosomes were isolated from all subjects. Isolated exosomes were then incubated with confluent endothelial cell monolayer cultures. Electric cell-substrate impedance sensing systems were used to determine the ability of exosomes to disrupt the intercellular barrier formed by confluent endothelial cells. In addition, immunofluorescent assessments of zonula occludens-1 tight junction protein cellular distribution were conducted to examine endothelial barrier dysfunction. miRNA and mRNA arrays were also applied to exosomes and endothelial cells, and miRNA inhibitors and mimics were transfected for mechanistic assays.
  
  
  
  Plasma exosomes isolated from either obese children or nonobese children with OSA were primarily derived from endothelial cell sources and recapitulated ED, or its absence, in naive human endothelial cells and also in vivo when injected into mice. Microarrays identified a restricted signature of exosomal miRNAs that readily distinguished ED from normal endothelial function. Among the miRNAs, expression of exosomal miRNA-630 was reduced in children with ED and normalized after therapy along with restoration of endothelial function. Conversely, transfection of exosomes from subjects without ED with an miRNA-630 inhibitor induces the ED functional phenotype. Gene target discovery experiments further revealed that miRNA-630 regulates 416 gene targets in endothelial cells that include the Nrf2, AMP kinase, and tight junction pathways.
  
  
  
  These observations elucidate a novel role of exosomal miRNA-360 as a putative key mediator of vascular function and cardiovascular disease risk in children with underlying OSA and/or obesity, and identify therapeutic targets.
authors.affiliation: 1 Section of Sleep Medicine, Department of Pediatrics, Pritzker School of Medicine, Biological Sciences Division, and.
authors.forename: Abdelnaby
authors.initials: A
authors.lastname: Khalyfa
authors.affiliation: 1 Section of Sleep Medicine, Department of Pediatrics, Pritzker School of Medicine, Biological Sciences Division, and.
authors.forename: Leila
authors.identifier: 0000-0003-3332-1057
authors.initials: L
authors.lastname: Kheirandish-Gozal
authors.affiliation: 1 Section of Sleep Medicine, Department of Pediatrics, Pritzker School of Medicine, Biological Sciences Division, and.
authors.forename: Ahamed A
authors.initials: AA
authors.lastname: Khalyfa
authors.affiliation: 1 Section of Sleep Medicine, Department of Pediatrics, Pritzker School of Medicine, Biological Sciences Division, and.
authors.forename: Mona F
authors.initials: MF
authors.lastname: Philby
authors.affiliation: 2 Sleep Unit, CIBER of Respiratory Diseases, Instituto Carlos III, CIBERES, Hospital Universitario de Burgos, Burgos, Spain.
authors.forename: María Luz
authors.initials: ML
authors.lastname: Alonso-Álvarez
authors.affiliation: 1 Section of Sleep Medicine, Department of Pediatrics, Pritzker School of Medicine, Biological Sciences Division, and.
authors.forename: Meelad
authors.initials: M
authors.lastname: Mohammadi
authors.affiliation: 1 Section of Sleep Medicine, Department of Pediatrics, Pritzker School of Medicine, Biological Sciences Division, and.
authors.forename: Rakesh
authors.initials: R
authors.lastname: Bhattacharjee
authors.affiliation: 2 Sleep Unit, CIBER of Respiratory Diseases, Instituto Carlos III, CIBERES, Hospital Universitario de Burgos, Burgos, Spain.
authors.forename: Joaquin
authors.initials: J
authors.lastname: Terán-Santos
authors.affiliation: 3 Center for Research Informatics, The University of Chicago, Chicago, Illinois; and.
authors.forename: Lei
authors.initials: L
authors.lastname: Huang
authors.affiliation: 3 Center for Research Informatics, The University of Chicago, Chicago, Illinois; and.
authors.forename: Jorge
authors.initials: J
authors.lastname: Andrade
authors.affiliation: 1 Section of Sleep Medicine, Department of Pediatrics, Pritzker School of Medicine, Biological Sciences Division, and.
authors.forename: David
authors.identifier: 0000-0001-8195-6036
authors.initials: D
authors.lastname: Gozal
mesh_terms: D016022:Case-Control Studies; D055252:Cell-Derived Microparticles / Q000378:metabolism*; D002648:Child; D004730:Endothelium, Vascular / Q000503:physiopathology*; D055354:Exosomes / Q000378:metabolism; D005260:Female; D005455:Fluorescent Antibody Technique; D006801:Humans; D008297:Male; D035683:MicroRNAs / Q000502:physiology*; D020411:Oligonucleotide Array Sequence Analysis; D063766:Pediatric Obesity / Q000150:complications / Q000503:physiopathology; D017286:Polysomnography; D020181:Sleep Apnea, Obstructive / Q000150:complications* / Q000503:physiopathology
chemical_list: C548399:MIRN630 microRNA, human; D035683:MicroRNAs
keywords: endothelial function; inflammation; intermittent hypoxia; obesity; sleep apnea