Mechanical stress induces pre-B-cell colony-enhancing factor/NAMPT expression via epigenetic regulation by miR-374a and miR-568 in human lung endothelium

Djanybek M. Adyshev, Venkateswaran Ramamoorthi Elangovan, Nurgul Moldobaeva, Brandon Mapes, Xiaoguang Sun, Joe G.N. Garcia

Research output: Contribution to journalArticle

31 Scopus citations

Abstract

Increased lung vascular permeability and alveolar edema are cardinal features of inflammatory conditions such as acute respiratory distress syndrome (ARDS) and ventilator-induced lung injury (VILI). We previously demonstrated that pre-B-cell colony-enhancing factor (PBEF)/NAMPT, the proinflammatory cytokine encoded by NAMPT, participates in ARDS and VILI inflammatory syndromes. The present study evaluated posttranscriptional regulation of PBEF/NAMPT gene expression in human lung endothelium via 39-untranslated region (UTR) microRNA (miRNA) binding. In silico analysis identified hsa-miR-374a and hsa-miR-568 as potential miRNA candidates. Increased PBEF/NAMPT transcription (by RT-PCR) and expression (byWestern blotting) induced by 18% cyclic stretch (CS) (2 h: 3.4 6 0.06 mRNA fold increase (FI); 10 h: 1.5 6 0.06 protein FI) and by LPS (4 h: 3.8 6 0.2 mRNA FI; 48 h: 2.6 6 0.2 protein FI) were significantly attenuated by transfection with mimics of hsa-miR-374a or hsa-miR-568 (40-60% reductions each). LPS and 18%CS increased the activity of a PBEF/NAMPT 39-UTR luciferase reporter (2.4-3.25 FI) with induction reduced by mimics of each miRNA (44-60% reduction). Specific miRNA inhibitors (antagomirs) for each PBEF/ NAMPT miRNA significantly increased the endogenous PBEF/ NAMPTmRNA(1.4-3.460.1 FI) and protein levels (1.2-1.460.1 FI) and 39-UTR luciferase activity (1.4-1.7 6 0.1 FI) compared with negative antagomir controls. Collectively, these data demonstrate that increased PBEF/NAMPT expression induced by bioactive agonists (i.e., excessive mechanical stress, LPS) involves epigenetic regulation with hsa-miR-374a and hsa-miR-568, representing novel therapeutic strategies to reduce inflammatory lung injury.

Original languageEnglish (US)
Pages (from-to)409-418
Number of pages10
JournalAmerican journal of respiratory cell and molecular biology
Volume50
Issue number2
DOIs
StatePublished - Feb 1 2014
Externally publishedYes

Keywords

  • ARDS
  • Endothelial cells
  • MIRNA
  • PBEF/NAMPT
  • Ventilator-induced lung injury

ASJC Scopus subject areas

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

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