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

TitleMechanical stress induces pre-B-cell colony-enhancing factor/NAMPT expression via epigenetic regulation by miR-374a and miR-568 in human lung endothelium.
Publication TypeJournal Article
Year of Publication2014
AuthorsAdyshev DM, Elangovan VRamamoorth, Moldobaeva N, Mapes B, Sun X, Garcia JGN
JournalAm J Respir Cell Mol Biol
Volume50
Issue2
Pagination409-18
Date Published2014 Feb
ISSN Number1535-4989
KeywordsCytokines, Endothelium, Epigenesis, Genetic, Gene Expression Regulation, Humans, Inflammation, Lipopolysaccharides, Lung, MicroRNAs, Nicotinamide Phosphoribosyltransferase, Respiratory Distress Syndrome, Adult, RNA, Messenger, Stress, Mechanical, Ventilator-Induced Lung Injury
Abstract

<p>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 3'-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 (by Western blotting) induced by 18% cyclic stretch (CS) (2 h: 3.4 ± 0.06 mRNA fold increase (FI); 10 h: 1.5 ± 0.06 protein FI) and by LPS (4 h: 3.8 ± 0.2 mRNA FI; 48 h: 2.6 ± 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 3'-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/NAMPT mRNA (1.4-3.4 ± 0.1 FI) and protein levels (1.2-1.4 ± 0.1 FI) and 3'-UTR luciferase activity (1.4-1.7 ± 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.</p>

DOI10.1165/rcmb.2013-0292OC
Alternate JournalAm. J. Respir. Cell Mol. Biol.
PubMed ID24053186
PubMed Central IDPMC3930953
Grant ListP01 HL98050 / HL / NHLBI NIH HHS / United States
R01 HL94394 / HL / NHLBI NIH HHS / United States
R01 HL094394 / HL / NHLBI NIH HHS / United States
P01 HL58064 / HL / NHLBI NIH HHS / United States
P01 HL098050 / HL / NHLBI NIH HHS / United States
P01 HL058064 / HL / NHLBI NIH HHS / United States