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 language | English (US) |
|---|---|
| Pages (from-to) | 409-418 |
| Number of pages | 10 |
| Journal | American Journal of Respiratory Cell and Molecular Biology |
| Volume | 50 |
| Issue number | 2 |
| DOIs | |
| State | Published - Feb 2014 |
| Externally published | Yes |
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Keywords
- ARDS
- Endothelial cells
- MIRNA
- PBEF/NAMPT
- Ventilator-induced lung injury
ASJC Scopus subject areas
- Cell Biology
- Pulmonary and Respiratory Medicine
- Molecular Biology
- Clinical Biochemistry
Cite this
Mechanical stress induces pre-B-cell colony-enhancing factor/NAMPT expression via epigenetic regulation by miR-374a and miR-568 in human lung endothelium. / Adyshev, Djanybek M.; Ramamoorthi Elangovan, Venkateswaran; Moldobaeva, Nurgul; Mapes, Brandon; Sun, Xiaoguang; Garcia, Joe GN.
In: American Journal of Respiratory Cell and Molecular Biology, Vol. 50, No. 2, 02.2014, p. 409-418.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Mechanical stress induces pre-B-cell colony-enhancing factor/NAMPT expression via epigenetic regulation by miR-374a and miR-568 in human lung endothelium
AU - Adyshev, Djanybek M.
AU - Ramamoorthi Elangovan, Venkateswaran
AU - Moldobaeva, Nurgul
AU - Mapes, Brandon
AU - Sun, Xiaoguang
AU - Garcia, Joe GN
PY - 2014/2
Y1 - 2014/2
N2 - 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.
AB - 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.
KW - ARDS
KW - Endothelial cells
KW - MIRNA
KW - PBEF/NAMPT
KW - Ventilator-induced lung injury
UR - http://www.scopus.com/inward/record.url?scp=84893231063&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84893231063&partnerID=8YFLogxK
U2 - 10.1165/rcmb.2013-0292OC
DO - 10.1165/rcmb.2013-0292OC
M3 - Article
C2 - 24053186
AN - SCOPUS:84893231063
VL - 50
SP - 409
EP - 418
JO - American Journal of Respiratory Cell and Molecular Biology
JF - American Journal of Respiratory Cell and Molecular Biology
SN - 1044-1549
IS - 2
ER -