title: Transcription factors and miRNAs that regulate fetal to adult CFTR expression change are new targets for cystic fibrosis.
abstract: The CFTR gene displays a tightly regulated tissue-specific and temporal expression. Mutations in this gene cause cystic fibrosis (CF). In this study we wanted to identify trans-regulatory elements responsible for CFTR differential expression in fetal and adult lung, and to determine the importance of inhibitory motifs in the CFTR-3'UTR with the aim of developing new tools for the correction of disease-causing mutations within CFTR. We show that lung development-specific transcription factors (FOXA, C/EBP) and microRNAs (miR-101, miR-145, miR-384) regulate the switch from strong fetal to very low CFTR expression after birth. By using miRNome profiling and gene reporter assays, we found that miR-101 and miR-145 are specifically upregulated in adult lung and that miR-101 directly acts on its cognate site in the CFTR-3'UTR in combination with an overlapping AU-rich element. We then designed miRNA-binding blocker oligonucleotides (MBBOs) to prevent binding of several miRNAs to the CFTR-3'UTR and tested them in primary human nasal epithelial cells from healthy individuals and CF patients carrying the p.Phe508del CFTR mutation. These MBBOs rescued CFTR channel activity by increasing CFTR mRNA and protein levels. Our data offer new understanding of the control of the CFTR gene regulation and new putative correctors for cystic fibrosis.
authors.affiliation: INSERM U827, Laboratoire de Génétique de Maladies Rares, Montpellier, France Université Montpellier I, UFR de Médecine, Montpellier, France Laboratoire de Génétique Moléculaire, CHU Montpellier, Montpellier, France.
authors.forename: Victoria
authors.initials: V
authors.lastname: Viart
authors.affiliation: INSERM U827, Laboratoire de Génétique de Maladies Rares, Montpellier, France Laboratoire de Génétique Moléculaire, CHU Montpellier, Montpellier, France.
authors.forename: Anne
authors.initials: A
authors.lastname: Bergougnoux
authors.affiliation: INSERM U827, Laboratoire de Génétique de Maladies Rares, Montpellier, France Université Montpellier I, UFR de Médecine, Montpellier, France.
authors.forename: Jennifer
authors.initials: J
authors.lastname: Bonini
authors.affiliation: INSERM U827, Laboratoire de Génétique de Maladies Rares, Montpellier, France Laboratoire de Génétique Moléculaire, CHU Montpellier, Montpellier, France.
authors.forename: Jessica
authors.initials: J
authors.lastname: Varilh
authors.affiliation: Centre de Ressources et de Compétences de la Mucoviscidose (CRCM), CHU Montpellier, Montpellier, France.
authors.forename: Raphaël
authors.initials: R
authors.lastname: Chiron
authors.affiliation: CDR St Antoine, INSERM UMR-S 938, Paris, France UPMC Université Paris 06, Paris, France.
authors.forename: Olivier
authors.initials: O
authors.lastname: Tabary
authors.affiliation: Département d'Information Médicale, CHU Montpellier, Montpellier, France UMR 729 MISTEA, Université Montpellier I, Montpellier, France.
authors.forename: Nicolas
authors.initials: N
authors.lastname: Molinari
authors.affiliation: INSERM U827, Laboratoire de Génétique de Maladies Rares, Montpellier, France Université Montpellier I, UFR de Médecine, Montpellier, France Laboratoire de Génétique Moléculaire, CHU Montpellier, Montpellier, France.
authors.forename: Mireille
authors.initials: M
authors.lastname: Claustres
authors.affiliation: INSERM U827, Laboratoire de Génétique de Maladies Rares, Montpellier, France Université Montpellier I, UFR de Médecine, Montpellier, France magali.taulan@inserm.fr.
authors.forename: Magali
authors.initials: M
authors.lastname: Taulan-Cadars
mesh_terms: D020413:3' Untranslated Regions; D000328:Adult; D000818:Animals; D001665:Binding Sites; D001980:Bronchi / Q000378:metabolism; D002460:Cell Line; D045744:Cell Line, Tumor; D047369:Chromatin Immunoprecipitation; D003550:Cystic Fibrosis / Q000188:drug therapy / Q000378:metabolism*; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000235:genetics / Q000378:metabolism*; D005260:Female; D020869:Gene Expression Profiling; D018507:Gene Expression Regulation, Developmental; D017930:Genes, Reporter; D006801:Humans; D008297:Male; D051379:Mice; D035683:MicroRNAs / Q000378:metabolism*; D016296:Mutagenesis; D009154:Mutation; D009841:Oligonucleotides / Q000737:chemistry; D014157:Transcription Factors / Q000378:metabolism
chemical_list: D020413:3' Untranslated Regions; C505032:CFTR protein, human; C534072:MIRN101 microRNA, human; C541010:MIRN145 microRNA, human; D035683:MicroRNAs; D009841:Oligonucleotides; D014157:Transcription Factors; D019005:Cystic Fibrosis Transmembrane Conductance Regulator