Limits...
miR-185 plays an anti-hypertrophic role in the heart via multiple targets in the calcium-signaling pathways.

Kim JO, Song DW, Kwon EJ, Hong SE, Song HK, Min CK, Kim do H - PLoS ONE (2015)

Bottom Line: The results showed that up-regulation of miR-185 led to anti-hypertrophic effects, while down-regulation led to pro-hypertrophic effects, suggesting that miR-185 has an anti-hypertrophic role in the heart.The expression of phospho-phospholamban (Thr-17), a marker of CaMKIIδ activity, was also significantly reduced by miR-185.In conclusion, miR-185 effectively blocked cardiac hypertrophy signaling through multiple targets, rendering it a potential drug target for diseases such as heart failure.

View Article: PubMed Central - PubMed

Affiliation: School of Life Sciences and Systems Biology Research Center, Gwangju Institute of Science and Technology (GIST), Gwangju, Korea.

ABSTRACT
MicroRNA (miRNA) is an endogenous non-coding RNA species that either inhibits RNA translation or promotes degradation of target mRNAs. miRNAs often regulate cellular signaling by targeting multiple genes within the pathways. In the present study, using Gene Set Analysis, a useful bioinformatics tool to identify miRNAs with multiple target genes in the same pathways, we identified miR-185 as a key candidate regulator of cardiac hypertrophy. Using a mouse model, we found that miR-185 was significantly down-regulated in myocardial cells during cardiac hypertrophy induced by transverse aortic constriction. To confirm that miR-185 is an anti-hypertrophic miRNA, genetic manipulation studies such as overexpression and knock-down of miR-185 in neonatal rat ventricular myocytes were conducted. The results showed that up-regulation of miR-185 led to anti-hypertrophic effects, while down-regulation led to pro-hypertrophic effects, suggesting that miR-185 has an anti-hypertrophic role in the heart. Our study further identified Camk2d, Ncx1, and Nfatc3 as direct targets of miR-185. The activity of Nuclear Factor of Activated T-cell (NFAT) and calcium/calmodulin-dependent protein kinase II delta (CaMKIIδ) was negatively regulated by miR-185 as assessed by NFAT-luciferase activity and western blotting. The expression of phospho-phospholamban (Thr-17), a marker of CaMKIIδ activity, was also significantly reduced by miR-185. In conclusion, miR-185 effectively blocked cardiac hypertrophy signaling through multiple targets, rendering it a potential drug target for diseases such as heart failure.

Show MeSH

Related in: MedlinePlus

miR-185 negatively regulate cardiomyocyte hypertrophy. (A, E) Microscopic images of immunofluorescence staining of NRVMs with α-actinin antibody. Twenty-four hours after transfection with miR-185 (15 nmol/L) (A) or miR-185 inhibitor (100 nmol/L) (E), NRVMs were stimulated with ET-1 for 48 h. Scale bar = 50 μm. (B, F) Cell surface areas of NRVMs measured using Image J software (N = 50 cells for each group). Untreated NRVMs served as controls. (C, D, G, H) qRT-PCR analysis of hypertrophic marker genes after transfection of miR-185 mimic (C, D) or miR-185 inhibitor (G, H) in the presence or absence of ET-1 (10 nmol/L). All data are expressed as mean ± SEM; *P < 0.05, **P < 0.001; N = 4.
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4358957&req=5

pone.0122509.g001: miR-185 negatively regulate cardiomyocyte hypertrophy. (A, E) Microscopic images of immunofluorescence staining of NRVMs with α-actinin antibody. Twenty-four hours after transfection with miR-185 (15 nmol/L) (A) or miR-185 inhibitor (100 nmol/L) (E), NRVMs were stimulated with ET-1 for 48 h. Scale bar = 50 μm. (B, F) Cell surface areas of NRVMs measured using Image J software (N = 50 cells for each group). Untreated NRVMs served as controls. (C, D, G, H) qRT-PCR analysis of hypertrophic marker genes after transfection of miR-185 mimic (C, D) or miR-185 inhibitor (G, H) in the presence or absence of ET-1 (10 nmol/L). All data are expressed as mean ± SEM; *P < 0.05, **P < 0.001; N = 4.

Mentions: Among the identified 18 miRNAs through GSA, six were previously unknown. We attempted to verify the effects of these six miRNAs (miR-185, miR-139–5p, miR-374, miR-324–5p, miR-153, and miR-141) on myocardial hypertrophy. qRT-PCR analysis of the hypertrophy markers ANF and BNP in transfected NRVMs showed that miR-139–5p and miR-374 mimics markedly increased the expression of ANF and BNP, while miR-324–5p, miR-153, and miR-141 mimics did not significantly affect the expression (unpublished data). However, transfection of miR-185 mimic significantly reduced the mRNA expression of ANF and BNP in NRVMs (Fig. 1C and 1D).


miR-185 plays an anti-hypertrophic role in the heart via multiple targets in the calcium-signaling pathways.

Kim JO, Song DW, Kwon EJ, Hong SE, Song HK, Min CK, Kim do H - PLoS ONE (2015)

miR-185 negatively regulate cardiomyocyte hypertrophy. (A, E) Microscopic images of immunofluorescence staining of NRVMs with α-actinin antibody. Twenty-four hours after transfection with miR-185 (15 nmol/L) (A) or miR-185 inhibitor (100 nmol/L) (E), NRVMs were stimulated with ET-1 for 48 h. Scale bar = 50 μm. (B, F) Cell surface areas of NRVMs measured using Image J software (N = 50 cells for each group). Untreated NRVMs served as controls. (C, D, G, H) qRT-PCR analysis of hypertrophic marker genes after transfection of miR-185 mimic (C, D) or miR-185 inhibitor (G, H) in the presence or absence of ET-1 (10 nmol/L). All data are expressed as mean ± SEM; *P < 0.05, **P < 0.001; N = 4.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4358957&req=5

pone.0122509.g001: miR-185 negatively regulate cardiomyocyte hypertrophy. (A, E) Microscopic images of immunofluorescence staining of NRVMs with α-actinin antibody. Twenty-four hours after transfection with miR-185 (15 nmol/L) (A) or miR-185 inhibitor (100 nmol/L) (E), NRVMs were stimulated with ET-1 for 48 h. Scale bar = 50 μm. (B, F) Cell surface areas of NRVMs measured using Image J software (N = 50 cells for each group). Untreated NRVMs served as controls. (C, D, G, H) qRT-PCR analysis of hypertrophic marker genes after transfection of miR-185 mimic (C, D) or miR-185 inhibitor (G, H) in the presence or absence of ET-1 (10 nmol/L). All data are expressed as mean ± SEM; *P < 0.05, **P < 0.001; N = 4.
Mentions: Among the identified 18 miRNAs through GSA, six were previously unknown. We attempted to verify the effects of these six miRNAs (miR-185, miR-139–5p, miR-374, miR-324–5p, miR-153, and miR-141) on myocardial hypertrophy. qRT-PCR analysis of the hypertrophy markers ANF and BNP in transfected NRVMs showed that miR-139–5p and miR-374 mimics markedly increased the expression of ANF and BNP, while miR-324–5p, miR-153, and miR-141 mimics did not significantly affect the expression (unpublished data). However, transfection of miR-185 mimic significantly reduced the mRNA expression of ANF and BNP in NRVMs (Fig. 1C and 1D).

Bottom Line: The results showed that up-regulation of miR-185 led to anti-hypertrophic effects, while down-regulation led to pro-hypertrophic effects, suggesting that miR-185 has an anti-hypertrophic role in the heart.The expression of phospho-phospholamban (Thr-17), a marker of CaMKIIδ activity, was also significantly reduced by miR-185.In conclusion, miR-185 effectively blocked cardiac hypertrophy signaling through multiple targets, rendering it a potential drug target for diseases such as heart failure.

View Article: PubMed Central - PubMed

Affiliation: School of Life Sciences and Systems Biology Research Center, Gwangju Institute of Science and Technology (GIST), Gwangju, Korea.

ABSTRACT
MicroRNA (miRNA) is an endogenous non-coding RNA species that either inhibits RNA translation or promotes degradation of target mRNAs. miRNAs often regulate cellular signaling by targeting multiple genes within the pathways. In the present study, using Gene Set Analysis, a useful bioinformatics tool to identify miRNAs with multiple target genes in the same pathways, we identified miR-185 as a key candidate regulator of cardiac hypertrophy. Using a mouse model, we found that miR-185 was significantly down-regulated in myocardial cells during cardiac hypertrophy induced by transverse aortic constriction. To confirm that miR-185 is an anti-hypertrophic miRNA, genetic manipulation studies such as overexpression and knock-down of miR-185 in neonatal rat ventricular myocytes were conducted. The results showed that up-regulation of miR-185 led to anti-hypertrophic effects, while down-regulation led to pro-hypertrophic effects, suggesting that miR-185 has an anti-hypertrophic role in the heart. Our study further identified Camk2d, Ncx1, and Nfatc3 as direct targets of miR-185. The activity of Nuclear Factor of Activated T-cell (NFAT) and calcium/calmodulin-dependent protein kinase II delta (CaMKIIδ) was negatively regulated by miR-185 as assessed by NFAT-luciferase activity and western blotting. The expression of phospho-phospholamban (Thr-17), a marker of CaMKIIδ activity, was also significantly reduced by miR-185. In conclusion, miR-185 effectively blocked cardiac hypertrophy signaling through multiple targets, rendering it a potential drug target for diseases such as heart failure.

Show MeSH
Related in: MedlinePlus