MicroRNA regulation of nonmuscle myosin light chain kinase expression in human lung endothelium

Djanybek M. Adyshev, Nurgul Moldobaeva, Brandon Mapes, Venkate Elangovan, Joe GN Garcia

Research output: Contribution to journalArticle

28 Citations (Scopus)

Abstract

Increased lung vascular permeability, the consequence of endothelial cell (EC) barrier dysfunction, is a cardinal feature of inflammatory conditions such as acute lung injury and sepsis and leads to lethal physiological dysfunction characterized by alveolar flooding, hypoxemia, and pulmonary edema. We previously demonstrated that the nonmuscle myosin light chain kinase isoform (nmMLCK) plays a key role in agonist-induced pulmonary EC barrier regulation. The present study evaluated posttranscriptional regulation of MYLK expression, the gene encoding nmMLCK, via 39 untranslated region (UTR) binding by microRNAs (miRNAs) with in silico analysis identifying hsa-miR-374a, hsa-miR-374b, hsa-miR-520c-3p, and hsa-miR-1290 as miRNA candidates. We identified increased MYLK gene transcription induced by TNF-α (24 h; 4.7 6 0.45 fold increase [FI]), LPS (4 h; 2.8560.15 [FI]), and 18% cyclic stretch (24 h; 4.660.24 FI) that was attenuated by transfection of human lung ECs with mimics of hsa-miR-374a, hsa-miR-374b, hsa-miR-520c-3p, or hsa-miR-1290 (20-80% reductions by each miRNA). TNF-α, LPS, and 18% cyclic stretch each increased the activity of a MYLK 39UTR luciferase reporter (2.5-7.0 FI) with induction reduced by mimics of each miRNA (30-60% reduction). MiRNA inhibitors (antagomirs) for each MYLK miRNA significantly increased 39UTR luciferase activity (1.2-2.3 FI) and rescued the decreased MLCK-39UTR reporter activity produced by miRNA mimics (70-110% increases for each miRNA; P , 0.05). These data demonstrate that increased human lung EC expression of MYLK by bioactive agonists (excessive mechanical stress, LPS, TNF-α) is regulated in part by specific miRNAs (hsa-miR-374a, hsamiR-374b, hsa-miR-520c-3p, and hsa-miR-1290), representing a novel therapeutic strategy for reducing inflammatory lung injury.

Original languageEnglish (US)
Pages (from-to)58-66
Number of pages9
JournalAmerican Journal of Respiratory Cell and Molecular Biology
Volume49
Issue number1
DOIs
StatePublished - Jul 2013
Externally publishedYes

Fingerprint

Myosin-Light-Chain Kinase
MicroRNAs
Endothelium
Lung
Endothelial cells
Endothelial Cells
Luciferases
Protein Isoforms
Untranslated Regions
Mechanical Stress
Gene encoding
Acute Lung Injury
Gene Expression Regulation
Capillary Permeability
Lung Injury
Pulmonary Edema
Transcription
Computer Simulation
Transfection
Sepsis

Keywords

  • Acute lung injury
  • Endothelial cells
  • MiRNA
  • MLCK
  • Ventilator-induced lung injury

ASJC Scopus subject areas

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

Cite this

MicroRNA regulation of nonmuscle myosin light chain kinase expression in human lung endothelium. / Adyshev, Djanybek M.; Moldobaeva, Nurgul; Mapes, Brandon; Elangovan, Venkate; Garcia, Joe GN.

In: American Journal of Respiratory Cell and Molecular Biology, Vol. 49, No. 1, 07.2013, p. 58-66.

Research output: Contribution to journalArticle

Adyshev, Djanybek M. ; Moldobaeva, Nurgul ; Mapes, Brandon ; Elangovan, Venkate ; Garcia, Joe GN. / MicroRNA regulation of nonmuscle myosin light chain kinase expression in human lung endothelium. In: American Journal of Respiratory Cell and Molecular Biology. 2013 ; Vol. 49, No. 1. pp. 58-66.
@article{49bbd80e3e724077a388a966fbe7cbb8,
title = "MicroRNA regulation of nonmuscle myosin light chain kinase expression in human lung endothelium",
abstract = "Increased lung vascular permeability, the consequence of endothelial cell (EC) barrier dysfunction, is a cardinal feature of inflammatory conditions such as acute lung injury and sepsis and leads to lethal physiological dysfunction characterized by alveolar flooding, hypoxemia, and pulmonary edema. We previously demonstrated that the nonmuscle myosin light chain kinase isoform (nmMLCK) plays a key role in agonist-induced pulmonary EC barrier regulation. The present study evaluated posttranscriptional regulation of MYLK expression, the gene encoding nmMLCK, via 39 untranslated region (UTR) binding by microRNAs (miRNAs) with in silico analysis identifying hsa-miR-374a, hsa-miR-374b, hsa-miR-520c-3p, and hsa-miR-1290 as miRNA candidates. We identified increased MYLK gene transcription induced by TNF-α (24 h; 4.7 6 0.45 fold increase [FI]), LPS (4 h; 2.8560.15 [FI]), and 18{\%} cyclic stretch (24 h; 4.660.24 FI) that was attenuated by transfection of human lung ECs with mimics of hsa-miR-374a, hsa-miR-374b, hsa-miR-520c-3p, or hsa-miR-1290 (20-80{\%} reductions by each miRNA). TNF-α, LPS, and 18{\%} cyclic stretch each increased the activity of a MYLK 39UTR luciferase reporter (2.5-7.0 FI) with induction reduced by mimics of each miRNA (30-60{\%} reduction). MiRNA inhibitors (antagomirs) for each MYLK miRNA significantly increased 39UTR luciferase activity (1.2-2.3 FI) and rescued the decreased MLCK-39UTR reporter activity produced by miRNA mimics (70-110{\%} increases for each miRNA; P , 0.05). These data demonstrate that increased human lung EC expression of MYLK by bioactive agonists (excessive mechanical stress, LPS, TNF-α) is regulated in part by specific miRNAs (hsa-miR-374a, hsamiR-374b, hsa-miR-520c-3p, and hsa-miR-1290), representing a novel therapeutic strategy for reducing inflammatory lung injury.",
keywords = "Acute lung injury, Endothelial cells, MiRNA, MLCK, Ventilator-induced lung injury",
author = "Adyshev, {Djanybek M.} and Nurgul Moldobaeva and Brandon Mapes and Venkate Elangovan and Garcia, {Joe GN}",
year = "2013",
month = "7",
doi = "10.1165/rcmb.2012-0397OC",
language = "English (US)",
volume = "49",
pages = "58--66",
journal = "American Journal of Respiratory Cell and Molecular Biology",
issn = "1044-1549",
publisher = "American Thoracic Society",
number = "1",

}

TY - JOUR

T1 - MicroRNA regulation of nonmuscle myosin light chain kinase expression in human lung endothelium

AU - Adyshev, Djanybek M.

AU - Moldobaeva, Nurgul

AU - Mapes, Brandon

AU - Elangovan, Venkate

AU - Garcia, Joe GN

PY - 2013/7

Y1 - 2013/7

N2 - Increased lung vascular permeability, the consequence of endothelial cell (EC) barrier dysfunction, is a cardinal feature of inflammatory conditions such as acute lung injury and sepsis and leads to lethal physiological dysfunction characterized by alveolar flooding, hypoxemia, and pulmonary edema. We previously demonstrated that the nonmuscle myosin light chain kinase isoform (nmMLCK) plays a key role in agonist-induced pulmonary EC barrier regulation. The present study evaluated posttranscriptional regulation of MYLK expression, the gene encoding nmMLCK, via 39 untranslated region (UTR) binding by microRNAs (miRNAs) with in silico analysis identifying hsa-miR-374a, hsa-miR-374b, hsa-miR-520c-3p, and hsa-miR-1290 as miRNA candidates. We identified increased MYLK gene transcription induced by TNF-α (24 h; 4.7 6 0.45 fold increase [FI]), LPS (4 h; 2.8560.15 [FI]), and 18% cyclic stretch (24 h; 4.660.24 FI) that was attenuated by transfection of human lung ECs with mimics of hsa-miR-374a, hsa-miR-374b, hsa-miR-520c-3p, or hsa-miR-1290 (20-80% reductions by each miRNA). TNF-α, LPS, and 18% cyclic stretch each increased the activity of a MYLK 39UTR luciferase reporter (2.5-7.0 FI) with induction reduced by mimics of each miRNA (30-60% reduction). MiRNA inhibitors (antagomirs) for each MYLK miRNA significantly increased 39UTR luciferase activity (1.2-2.3 FI) and rescued the decreased MLCK-39UTR reporter activity produced by miRNA mimics (70-110% increases for each miRNA; P , 0.05). These data demonstrate that increased human lung EC expression of MYLK by bioactive agonists (excessive mechanical stress, LPS, TNF-α) is regulated in part by specific miRNAs (hsa-miR-374a, hsamiR-374b, hsa-miR-520c-3p, and hsa-miR-1290), representing a novel therapeutic strategy for reducing inflammatory lung injury.

AB - Increased lung vascular permeability, the consequence of endothelial cell (EC) barrier dysfunction, is a cardinal feature of inflammatory conditions such as acute lung injury and sepsis and leads to lethal physiological dysfunction characterized by alveolar flooding, hypoxemia, and pulmonary edema. We previously demonstrated that the nonmuscle myosin light chain kinase isoform (nmMLCK) plays a key role in agonist-induced pulmonary EC barrier regulation. The present study evaluated posttranscriptional regulation of MYLK expression, the gene encoding nmMLCK, via 39 untranslated region (UTR) binding by microRNAs (miRNAs) with in silico analysis identifying hsa-miR-374a, hsa-miR-374b, hsa-miR-520c-3p, and hsa-miR-1290 as miRNA candidates. We identified increased MYLK gene transcription induced by TNF-α (24 h; 4.7 6 0.45 fold increase [FI]), LPS (4 h; 2.8560.15 [FI]), and 18% cyclic stretch (24 h; 4.660.24 FI) that was attenuated by transfection of human lung ECs with mimics of hsa-miR-374a, hsa-miR-374b, hsa-miR-520c-3p, or hsa-miR-1290 (20-80% reductions by each miRNA). TNF-α, LPS, and 18% cyclic stretch each increased the activity of a MYLK 39UTR luciferase reporter (2.5-7.0 FI) with induction reduced by mimics of each miRNA (30-60% reduction). MiRNA inhibitors (antagomirs) for each MYLK miRNA significantly increased 39UTR luciferase activity (1.2-2.3 FI) and rescued the decreased MLCK-39UTR reporter activity produced by miRNA mimics (70-110% increases for each miRNA; P , 0.05). These data demonstrate that increased human lung EC expression of MYLK by bioactive agonists (excessive mechanical stress, LPS, TNF-α) is regulated in part by specific miRNAs (hsa-miR-374a, hsamiR-374b, hsa-miR-520c-3p, and hsa-miR-1290), representing a novel therapeutic strategy for reducing inflammatory lung injury.

KW - Acute lung injury

KW - Endothelial cells

KW - MiRNA

KW - MLCK

KW - Ventilator-induced lung injury

UR - http://www.scopus.com/inward/record.url?scp=84883172305&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84883172305&partnerID=8YFLogxK

U2 - 10.1165/rcmb.2012-0397OC

DO - 10.1165/rcmb.2012-0397OC

M3 - Article

C2 - 23492194

AN - SCOPUS:84883172305

VL - 49

SP - 58

EP - 66

JO - American Journal of Respiratory Cell and Molecular Biology

JF - American Journal of Respiratory Cell and Molecular Biology

SN - 1044-1549

IS - 1

ER -