Loss of microRNA-17-92 in smooth muscle cells attenuates experimental pulmonary hypertension via induction of PDZ and LIM domain 5

Tianji Chen, Guofei Zhou, Qiyuan Zhou, Haiyang Tang, Joyce Christina F Ibe, Hongqiang Cheng, Deming Gou, Ju Chen, Jason Yuan, J. Usha Raj

Research output: Contribution to journalArticle

33 Citations (Scopus)

Abstract

Rationale: Recent studies suggest that microRNAs (miRNAs) play important roles in regulation of pulmonary artery smooth muscle cell (PASMC) phenotype and are implicated in pulmonary arterial hypertension (PAH). However, the underlying molecular mechanisms remain elusive. Objectives: This study aims to understand the mechanisms regulating PASMC proliferation and differentiation by microRNA-17-92 (miR-17-92) and to elucidate its implication in PAH. Methods: We generated smooth muscle cell (SMC)-specific miR-17-92 and PDZ and LIM domain 5 (PDLIM5) knockout mice and overexpressed miR-17-92 and PDLIM5 by injection of miR-17-92 mimics or PDLIM5-V5-His plasmids and measured their responses to hypoxia. We used miR-17-92 mimics, inhibitors, overexpression vectors, small interfering RNAs against PDLIM5, Smad, and transforming growth factor (TGF)-β to determine the role of miR-17-92 and its downstream targets in PASMC proliferation and differentiation. Measurements and Main Results: Wefound that human PASMC (HPASMC) from patients with PAH expressed decreased levels of the miR-17-92 cluster, TGF-β, and SMC markers. Overexpression of miR-17-92 increased and restored the expression of TGF-β3, Smad3, and SMC markers in HPASMC of normal subjects and patients with idiopathic PAH, respectively. Knockdown of Smad3 but not Smad2 prevented miR-17-92-induced expression of SMC markers. SMC-specific knockout of miR-17-92 attenuated hypoxia-induced pulmonary hypertension (PH) in mice, whereas reconstitution of miR-17-92 restored hypoxia-induced PH in these mice. We also found that PDLIM5 is a direct target of miR-17/20a, and hypertensive HPASMC and mouse PASMC expressed elevated PDLIM5 levels. Suppression of PDLIM5 increased expression of SMC markers and enhanced TGF-β/Smad2/3 activity in vitro and enhanced hypoxia-induced PH in vivo, whereas overexpression of PDLIM5 attenuated hypoxia-induced PH. Conclusions: We provided the first evidence that miR-17-92 inhibits PDLIM5 to induce the TGF-β3/SMAD3 pathway, contributing to the pathogenesis of PAH.

Original languageEnglish (US)
Pages (from-to)678-692
Number of pages15
JournalAmerican Journal of Respiratory and Critical Care Medicine
Volume191
Issue number6
DOIs
StatePublished - Mar 15 2015

Fingerprint

PDZ Domains
MicroRNAs
Pulmonary Hypertension
Smooth Muscle Myocytes
Transforming Growth Factors
Pulmonary Artery
Cell Differentiation
Cell Proliferation
Knockout Mice
Small Interfering RNA

Keywords

  • MicroRNA-17-92
  • PDLIM5
  • Pulmonary arterial hypertension
  • SMAD3
  • Transforming growth factor-β3

ASJC Scopus subject areas

  • Pulmonary and Respiratory Medicine
  • Critical Care and Intensive Care Medicine

Cite this

Loss of microRNA-17-92 in smooth muscle cells attenuates experimental pulmonary hypertension via induction of PDZ and LIM domain 5. / Chen, Tianji; Zhou, Guofei; Zhou, Qiyuan; Tang, Haiyang; Ibe, Joyce Christina F; Cheng, Hongqiang; Gou, Deming; Chen, Ju; Yuan, Jason; Raj, J. Usha.

In: American Journal of Respiratory and Critical Care Medicine, Vol. 191, No. 6, 15.03.2015, p. 678-692.

Research output: Contribution to journalArticle

Chen, Tianji ; Zhou, Guofei ; Zhou, Qiyuan ; Tang, Haiyang ; Ibe, Joyce Christina F ; Cheng, Hongqiang ; Gou, Deming ; Chen, Ju ; Yuan, Jason ; Raj, J. Usha. / Loss of microRNA-17-92 in smooth muscle cells attenuates experimental pulmonary hypertension via induction of PDZ and LIM domain 5. In: American Journal of Respiratory and Critical Care Medicine. 2015 ; Vol. 191, No. 6. pp. 678-692.
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abstract = "Rationale: Recent studies suggest that microRNAs (miRNAs) play important roles in regulation of pulmonary artery smooth muscle cell (PASMC) phenotype and are implicated in pulmonary arterial hypertension (PAH). However, the underlying molecular mechanisms remain elusive. Objectives: This study aims to understand the mechanisms regulating PASMC proliferation and differentiation by microRNA-17-92 (miR-17-92) and to elucidate its implication in PAH. Methods: We generated smooth muscle cell (SMC)-specific miR-17-92 and PDZ and LIM domain 5 (PDLIM5) knockout mice and overexpressed miR-17-92 and PDLIM5 by injection of miR-17-92 mimics or PDLIM5-V5-His plasmids and measured their responses to hypoxia. We used miR-17-92 mimics, inhibitors, overexpression vectors, small interfering RNAs against PDLIM5, Smad, and transforming growth factor (TGF)-β to determine the role of miR-17-92 and its downstream targets in PASMC proliferation and differentiation. Measurements and Main Results: Wefound that human PASMC (HPASMC) from patients with PAH expressed decreased levels of the miR-17-92 cluster, TGF-β, and SMC markers. Overexpression of miR-17-92 increased and restored the expression of TGF-β3, Smad3, and SMC markers in HPASMC of normal subjects and patients with idiopathic PAH, respectively. Knockdown of Smad3 but not Smad2 prevented miR-17-92-induced expression of SMC markers. SMC-specific knockout of miR-17-92 attenuated hypoxia-induced pulmonary hypertension (PH) in mice, whereas reconstitution of miR-17-92 restored hypoxia-induced PH in these mice. We also found that PDLIM5 is a direct target of miR-17/20a, and hypertensive HPASMC and mouse PASMC expressed elevated PDLIM5 levels. Suppression of PDLIM5 increased expression of SMC markers and enhanced TGF-β/Smad2/3 activity in vitro and enhanced hypoxia-induced PH in vivo, whereas overexpression of PDLIM5 attenuated hypoxia-induced PH. Conclusions: We provided the first evidence that miR-17-92 inhibits PDLIM5 to induce the TGF-β3/SMAD3 pathway, contributing to the pathogenesis of PAH.",
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AU - Chen, Tianji

AU - Zhou, Guofei

AU - Zhou, Qiyuan

AU - Tang, Haiyang

AU - Ibe, Joyce Christina F

AU - Cheng, Hongqiang

AU - Gou, Deming

AU - Chen, Ju

AU - Yuan, Jason

AU - Raj, J. Usha

PY - 2015/3/15

Y1 - 2015/3/15

N2 - Rationale: Recent studies suggest that microRNAs (miRNAs) play important roles in regulation of pulmonary artery smooth muscle cell (PASMC) phenotype and are implicated in pulmonary arterial hypertension (PAH). However, the underlying molecular mechanisms remain elusive. Objectives: This study aims to understand the mechanisms regulating PASMC proliferation and differentiation by microRNA-17-92 (miR-17-92) and to elucidate its implication in PAH. Methods: We generated smooth muscle cell (SMC)-specific miR-17-92 and PDZ and LIM domain 5 (PDLIM5) knockout mice and overexpressed miR-17-92 and PDLIM5 by injection of miR-17-92 mimics or PDLIM5-V5-His plasmids and measured their responses to hypoxia. We used miR-17-92 mimics, inhibitors, overexpression vectors, small interfering RNAs against PDLIM5, Smad, and transforming growth factor (TGF)-β to determine the role of miR-17-92 and its downstream targets in PASMC proliferation and differentiation. Measurements and Main Results: Wefound that human PASMC (HPASMC) from patients with PAH expressed decreased levels of the miR-17-92 cluster, TGF-β, and SMC markers. Overexpression of miR-17-92 increased and restored the expression of TGF-β3, Smad3, and SMC markers in HPASMC of normal subjects and patients with idiopathic PAH, respectively. Knockdown of Smad3 but not Smad2 prevented miR-17-92-induced expression of SMC markers. SMC-specific knockout of miR-17-92 attenuated hypoxia-induced pulmonary hypertension (PH) in mice, whereas reconstitution of miR-17-92 restored hypoxia-induced PH in these mice. We also found that PDLIM5 is a direct target of miR-17/20a, and hypertensive HPASMC and mouse PASMC expressed elevated PDLIM5 levels. Suppression of PDLIM5 increased expression of SMC markers and enhanced TGF-β/Smad2/3 activity in vitro and enhanced hypoxia-induced PH in vivo, whereas overexpression of PDLIM5 attenuated hypoxia-induced PH. Conclusions: We provided the first evidence that miR-17-92 inhibits PDLIM5 to induce the TGF-β3/SMAD3 pathway, contributing to the pathogenesis of PAH.

AB - Rationale: Recent studies suggest that microRNAs (miRNAs) play important roles in regulation of pulmonary artery smooth muscle cell (PASMC) phenotype and are implicated in pulmonary arterial hypertension (PAH). However, the underlying molecular mechanisms remain elusive. Objectives: This study aims to understand the mechanisms regulating PASMC proliferation and differentiation by microRNA-17-92 (miR-17-92) and to elucidate its implication in PAH. Methods: We generated smooth muscle cell (SMC)-specific miR-17-92 and PDZ and LIM domain 5 (PDLIM5) knockout mice and overexpressed miR-17-92 and PDLIM5 by injection of miR-17-92 mimics or PDLIM5-V5-His plasmids and measured their responses to hypoxia. We used miR-17-92 mimics, inhibitors, overexpression vectors, small interfering RNAs against PDLIM5, Smad, and transforming growth factor (TGF)-β to determine the role of miR-17-92 and its downstream targets in PASMC proliferation and differentiation. Measurements and Main Results: Wefound that human PASMC (HPASMC) from patients with PAH expressed decreased levels of the miR-17-92 cluster, TGF-β, and SMC markers. Overexpression of miR-17-92 increased and restored the expression of TGF-β3, Smad3, and SMC markers in HPASMC of normal subjects and patients with idiopathic PAH, respectively. Knockdown of Smad3 but not Smad2 prevented miR-17-92-induced expression of SMC markers. SMC-specific knockout of miR-17-92 attenuated hypoxia-induced pulmonary hypertension (PH) in mice, whereas reconstitution of miR-17-92 restored hypoxia-induced PH in these mice. We also found that PDLIM5 is a direct target of miR-17/20a, and hypertensive HPASMC and mouse PASMC expressed elevated PDLIM5 levels. Suppression of PDLIM5 increased expression of SMC markers and enhanced TGF-β/Smad2/3 activity in vitro and enhanced hypoxia-induced PH in vivo, whereas overexpression of PDLIM5 attenuated hypoxia-induced PH. Conclusions: We provided the first evidence that miR-17-92 inhibits PDLIM5 to induce the TGF-β3/SMAD3 pathway, contributing to the pathogenesis of PAH.

KW - MicroRNA-17-92

KW - PDLIM5

KW - Pulmonary arterial hypertension

KW - SMAD3

KW - Transforming growth factor-β3

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