title: Profibrotic role of miR-154 in pulmonary fibrosis.
abstract: In this study, we explored the regulation and the role of up-regulated microRNAs in idiopathic pulmonary fibrosis (IPF), a progressive interstitial lung disease of unknown origin. We analyzed the expression of microRNAs in IPF lungs and identified 43 significantly up-regulated microRNAs. Twenty-four of the 43 increased microRNAs were localized to the chromosome 14q32 microRNA cluster. We validated the increased expression of miR-154, miR-134, miR-299-5p, miR-410, miR-382, miR-409-3p, miR-487b, miR-31, and miR-127 by quantitative RT-PCR and determined that they were similarly expressed in embryonic lungs. We did not find evidence for differential methylation in this region, but analysis of transcription factor binding sites identified multiple SMAD3-binding elements in the 14q32 microRNA cluster. TGF-β1 stimulation of normal human lung fibroblasts (NHLF) caused up-regulation of microRNAs on chr14q32 that were also increased in IPF lungs. Chromatin immunoprecipitation confirmed binding of SMAD3 to the putative promoter of miR-154. Mir-154 was increased in IPF fibroblasts, and transfection of NHLF with miR-154 caused significant increases in cell proliferation and migration. The increase in proliferation induced by TGF-β was not observed when NHLF or IPF fibroblasts were transfected with a mir-154 inhibitor. Transfection with miR-154 caused activation of the WNT pathway in NHLF. ICG-001 and XAV939, inhibitors of the WNT/β-catenin pathway, reduced the proliferative effect of miR-154. The potential role of miR-154, one of multiple chr14q32 microRNA cluster members up-regulated in IPF and a regulator of fibroblast migration and proliferation, should be further explored in IPF.
authors.affiliation: Dorothy P. and Richard P. Simmons Center for Interstitial Lung Disease, Pulmonary, Allergy and Critical Care Medicine, University of Pittsburgh, Pittsburgh, PA 15261, USA.
authors.forename: Jadranka
authors.initials: J
authors.lastname: Milosevic
authors.forename: Kusum
authors.initials: K
authors.lastname: Pandit
authors.forename: Marcus
authors.initials: M
authors.lastname: Magister
authors.forename: Einat
authors.initials: E
authors.lastname: Rabinovich
authors.forename: Daniel C
authors.initials: DC
authors.lastname: Ellwanger
authors.forename: Guoying
authors.initials: G
authors.lastname: Yu
authors.forename: Louis J
authors.initials: LJ
authors.lastname: Vuga
authors.forename: Benny
authors.initials: B
authors.lastname: Weksler
authors.forename: Panayiotis V
authors.initials: PV
authors.lastname: Benos
authors.forename: Kevin F
authors.initials: KF
authors.lastname: Gibson
authors.forename: Michael
authors.initials: M
authors.lastname: McMillan
authors.forename: Michael
authors.initials: M
authors.lastname: Kahn
authors.forename: Naftali
authors.initials: N
authors.lastname: Kaminski
mesh_terms: D016022:Case-Control Studies; D002465:Cell Movement; D049109:Cell Proliferation; D002478:Cells, Cultured; D002883:Chromosomes, Human, Pair 14; D050762:Cyclin-Dependent Kinase Inhibitor p15 / Q000235:genetics / Q000378:metabolism; D005347:Fibroblasts / Q000378:metabolism / Q000473:pathology / Q000502:physiology; D015870:Gene Expression; D006801:Humans; D008168:Lung / Q000378:metabolism / Q000473:pathology; D035683:MicroRNAs / Q000502:physiology*; D005810:Multigene Family; D020411:Oligonucleotide Array Sequence Analysis; D011658:Pulmonary Fibrosis / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D034622:RNA Interference; D059467:Transcriptome; D053773:Transforming Growth Factor beta1 / Q000502:physiology; D060449:Wnt Signaling Pathway
chemical_list: D050762:Cyclin-Dependent Kinase Inhibitor p15; C561122:MIRN154 microRNA, human; D035683:MicroRNAs; D053773:Transforming Growth Factor beta1