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

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

Research output: Contribution to journalArticle

28 Scopus citations

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 1 2013
Externally publishedYes

Keywords

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

ASJC Scopus subject areas

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

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