Profibrotic role of miR-154 in pulmonary fibrosis

Jadranka Milosevic, Kusum Pandit, Marcus Magister, Einat Rabinovich, Daniel C. Ellwanger, Guoying Yu, Louis J. Vuga, Benny Weksler, Panayiotis V. Benos, Kevin F. Gibson, Michael McMillan, Michael Kahn, Naftali Kaminski

Research output: Contribution to journalArticle

105 Citations (Scopus)

Abstract

In this study, we explored the regulationandthe 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 upregulated 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 micro- RNA cluster members up-regulatedin IPF and a regulator of fibroblast migration and proliferation, should be further explored in IPF.

Original languageEnglish (US)
Pages (from-to)879-887
Number of pages9
JournalAmerican journal of respiratory cell and molecular biology
Volume47
Issue number6
DOIs
StatePublished - Dec 1 2012

Fingerprint

Pulmonary Fibrosis
Idiopathic Pulmonary Fibrosis
MicroRNAs
Fibroblasts
Lung
Transfection
Catenins
Pulmonary diseases
Methylation
Chromatin Immunoprecipitation
Interstitial Lung Diseases
Cell proliferation
Chromosomes
Chromatin
Cell Movement
Transcription Factors
Up-Regulation
Chemical activation
Binding Sites
Cell Proliferation

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Pulmonary and Respiratory Medicine
  • Clinical Biochemistry
  • Cell Biology

Cite this

Milosevic, J., Pandit, K., Magister, M., Rabinovich, E., Ellwanger, D. C., Yu, G., ... Kaminski, N. (2012). Profibrotic role of miR-154 in pulmonary fibrosis. American journal of respiratory cell and molecular biology, 47(6), 879-887. https://doi.org/10.1165/rcmb.2011-0377OC

Profibrotic role of miR-154 in pulmonary fibrosis. / Milosevic, Jadranka; Pandit, Kusum; Magister, Marcus; Rabinovich, Einat; Ellwanger, Daniel C.; Yu, Guoying; Vuga, Louis J.; Weksler, Benny; Benos, Panayiotis V.; Gibson, Kevin F.; McMillan, Michael; Kahn, Michael; Kaminski, Naftali.

In: American journal of respiratory cell and molecular biology, Vol. 47, No. 6, 01.12.2012, p. 879-887.

Research output: Contribution to journalArticle

Milosevic, J, Pandit, K, Magister, M, Rabinovich, E, Ellwanger, DC, Yu, G, Vuga, LJ, Weksler, B, Benos, PV, Gibson, KF, McMillan, M, Kahn, M & Kaminski, N 2012, 'Profibrotic role of miR-154 in pulmonary fibrosis', American journal of respiratory cell and molecular biology, vol. 47, no. 6, pp. 879-887. https://doi.org/10.1165/rcmb.2011-0377OC
Milosevic J, Pandit K, Magister M, Rabinovich E, Ellwanger DC, Yu G et al. Profibrotic role of miR-154 in pulmonary fibrosis. American journal of respiratory cell and molecular biology. 2012 Dec 1;47(6):879-887. https://doi.org/10.1165/rcmb.2011-0377OC
Milosevic, Jadranka ; Pandit, Kusum ; Magister, Marcus ; Rabinovich, Einat ; Ellwanger, Daniel C. ; Yu, Guoying ; Vuga, Louis J. ; Weksler, Benny ; Benos, Panayiotis V. ; Gibson, Kevin F. ; McMillan, Michael ; Kahn, Michael ; Kaminski, Naftali. / Profibrotic role of miR-154 in pulmonary fibrosis. In: American journal of respiratory cell and molecular biology. 2012 ; Vol. 47, No. 6. pp. 879-887.
@article{8a1c70b78fd540ed9abacdedbce4cca2,
title = "Profibrotic role of miR-154 in pulmonary fibrosis",
abstract = "In this study, we explored the regulationandthe 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 upregulated 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 micro- RNA cluster members up-regulatedin IPF and a regulator of fibroblast migration and proliferation, should be further explored in IPF.",
author = "Jadranka Milosevic and Kusum Pandit and Marcus Magister and Einat Rabinovich and Ellwanger, {Daniel C.} and Guoying Yu and Vuga, {Louis J.} and Benny Weksler and Benos, {Panayiotis V.} and Gibson, {Kevin F.} and Michael McMillan and Michael Kahn and Naftali Kaminski",
year = "2012",
month = "12",
day = "1",
doi = "10.1165/rcmb.2011-0377OC",
language = "English (US)",
volume = "47",
pages = "879--887",
journal = "American Journal of Respiratory Cell and Molecular Biology",
issn = "1044-1549",
publisher = "American Thoracic Society",
number = "6",

}

TY - JOUR

T1 - Profibrotic role of miR-154 in pulmonary fibrosis

AU - Milosevic, Jadranka

AU - Pandit, Kusum

AU - Magister, Marcus

AU - Rabinovich, Einat

AU - Ellwanger, Daniel C.

AU - Yu, Guoying

AU - Vuga, Louis J.

AU - Weksler, Benny

AU - Benos, Panayiotis V.

AU - Gibson, Kevin F.

AU - McMillan, Michael

AU - Kahn, Michael

AU - Kaminski, Naftali

PY - 2012/12/1

Y1 - 2012/12/1

N2 - In this study, we explored the regulationandthe 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 upregulated 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 micro- RNA cluster members up-regulatedin IPF and a regulator of fibroblast migration and proliferation, should be further explored in IPF.

AB - In this study, we explored the regulationandthe 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 upregulated 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 micro- RNA cluster members up-regulatedin IPF and a regulator of fibroblast migration and proliferation, should be further explored in IPF.

UR - http://www.scopus.com/inward/record.url?scp=84870508377&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84870508377&partnerID=8YFLogxK

U2 - 10.1165/rcmb.2011-0377OC

DO - 10.1165/rcmb.2011-0377OC

M3 - Article

VL - 47

SP - 879

EP - 887

JO - American Journal of Respiratory Cell and Molecular Biology

JF - American Journal of Respiratory Cell and Molecular Biology

SN - 1044-1549

IS - 6

ER -