title: Unique Tracheal Fluid MicroRNA Signature Predicts Response to FETO in Patients With Congenital Diaphragmatic Hernia.
abstract: OBJECTIVE
  Our objective was to determine the fetal in vivo microRNA signature in hypoplastic lungs of human fetuses with severe isolated congenital diaphragmatic hernia (CDH) and changes in tracheal and amniotic fluid of fetuses undergoing fetoscopic endoluminal tracheal occlusion (FETO) to reverse severe lung hypoplasia due to CDH.
  
  
  METHODS
  We profiled microRNA expression in prenatal human lungs by microarray analysis. We then validated this signature with real-time quantitative polymerase chain reaction in tracheal and amniotic fluid of CDH patients undergoing FETO. We further explored the role of miR-200b using semiquantitative in situ hybridization and immunohistochemistry for TGF-β2 in postnatal lung sections. We investigated miR-200b effects on TGF-β signaling using a SMAD-luciferase reporter assay and Western blotting for phospho-SMAD2/3 and ZEB-2 in cultures of human bronchial epithelial cells.
  
  
  RESULTS
  CDH lungs display an increased expression of 2 microRNAs: miR-200b and miR-10a as compared to control lungs. Fetuses undergoing FETO display increased miR-200 expression in their tracheal fluid at the time of balloon removal. Future survivors of FETO display significantly higher miR-200 expression than those with a limited response. miR-200b was expressed in bronchial epithelial cells and vascular endothelial cells. TGF-β2 expression was lower in CDH lungs. miR-200b inhibited TGF-β-induced SMAD signaling in cultures of human bronchial epithelial cells.
  
  
  CONCLUSIONS
  Human fetal hypoplastic CDH lungs have a specific miR-200/miR-10a signature. Survival after FETO is associated with increased miR-200 family expression. miR-200b overexpression in CDH lungs results in decreased TGF-β/SMAD signaling.
authors.affiliation: *Departments of Surgery, Division of Pediatric Surgery, Pediatrics & Child Health and Physiology & Pathophysiology, University of Manitoba and Biology of Breathing Theme, Manitoba Institute of Child Health, Winnipeg, Manitoba, Canada †Life and Health Sciences Research Institute/3B's-PT Government Associate Laboratory, Braga/Guimarães, Portugal and Department of Pediatric Surgery, Hospital de Braga, Braga, Portugal ‡Clinical Department of Obstetrics and Gynaecology, University Hospitals Leuven and Academic Department Development and Regeneration, Organ System Cluster, KU Leuven, Leuven, Belgium §Department of Pediatric Surgery, Erasmus MC-Sophia, Rotterdam, The Netherlands ¶Program in Physiology and Experimental Medicine, Hospital for Sick Children, Toronto, Ontario, Canada.
authors.forename: Patrícia
authors.initials: P
authors.lastname: Pereira-Terra
authors.forename: Jan A
authors.initials: JA
authors.lastname: Deprest
authors.forename: Ramin
authors.initials: R
authors.lastname: Kholdebarin
authors.forename: Naghmeh
authors.initials: N
authors.lastname: Khoshgoo
authors.forename: Philip
authors.initials: P
authors.lastname: DeKoninck
authors.forename: Anne A Boerema-De
authors.initials: AA
authors.lastname: Munck
authors.forename: Jinxia
authors.initials: J
authors.lastname: Wang
authors.forename: Fuqin
authors.initials: F
authors.lastname: Zhu
authors.forename: Robbert J
authors.initials: RJ
authors.lastname: Rottier
authors.forename: Barbara M
authors.initials: BM
authors.lastname: Iwasiow
authors.forename: Jorge
authors.initials: J
authors.lastname: Correia-Pinto
authors.forename: Dick
authors.initials: D
authors.lastname: Tibboel
authors.forename: Martin
authors.initials: M
authors.lastname: Post
authors.forename: Richard
authors.initials: R
authors.lastname: Keijzer
mesh_terms: D000653:Amniotic Fluid / Q000378:metabolism*; D015415:Biomarkers / Q000378:metabolism; D016022:Case-Control Studies; D005260:Female; D005332:Fetoscopy / Q000379:methods / Q000401:mortality; D005786:Gene Expression Regulation; D065630:Hernias, Diaphragmatic, Congenital / Q000235:genetics / Q000378:metabolism / Q000401:mortality / Q000628:therapy*; D006801:Humans; D007231:Infant, Newborn; D008168:Lung / Q000002:abnormalities / Q000378:metabolism*; D008297:Male; D035683:MicroRNAs / Q000378:metabolism*; D020411:Oligonucleotide Array Sequence Analysis; D011247:Pregnancy; D014132:Trachea / Q000378:metabolism*
chemical_list: D015415:Biomarkers; C547183:MIRN10 microRNA, human; C533896:MIRN200 microRNA, human; D035683:MicroRNAs