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Gene remodeling in type 2 diabetic cardiomyopathy and its phenotypic rescue with SERCA2a.

Karakikes I, Kim M, Hadri L, Sakata S, Sun Y, Zhang W, Chemaly ER, Hajjar RJ, Lebeche D - PLoS ONE (2009)

Bottom Line: Diabetes-associated myocardial dysfunction results in altered gene expression in the heart.Diabetic cardiomyopathic hearts have reduced levels of SERCA2a.In isolated cardiomyocytes in vitro, SERCA2a overexpression significantly modified the expression of a number of transcripts known to be involved in insulin signaling, glucose metabolism and cardiac remodeling.

View Article: PubMed Central - PubMed

Affiliation: Cardiovascular Research Center, Mount Sinai School of Medicine, New York, New York, United States of America.

ABSTRACT

Background/aim: Diabetes-associated myocardial dysfunction results in altered gene expression in the heart. We aimed to investigate the changes in gene expression profiles accompanying diabetes-induced cardiomyopathy and its phenotypic rescue by restoration of SERCA2a expression.

Methods/results: Using the Otsuka Long-Evans Tokushima Fatty rat model of type 2 diabetes and the Agilent rat microarray chip, we analyzed gene expression by comparing differential transcriptional changes in age-matched control versus diabetic hearts and diabetic hearts that received gene transfer of SERCA2a. Microarray expression profiles of selected genes were verified with real-time qPCR and immunoblotting. Our analysis indicates that diabetic cardiomyopathy is associated with a downregulation of transcripts. Diabetic cardiomyopathic hearts have reduced levels of SERCA2a. SERCA2a gene transfer in these hearts reduced diabetes-associated hypertrophy, and differentially modulated the expression of 76 genes and reversed the transcriptional profile induced by diabetes. In isolated cardiomyocytes in vitro, SERCA2a overexpression significantly modified the expression of a number of transcripts known to be involved in insulin signaling, glucose metabolism and cardiac remodeling.

Conclusion: This investigation provided insight into the pathophysiology of cardiac remodeling and the potential role of SERCA2a normalization in multiple pathways in diabetic cardiomyopathy.

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Related in: MedlinePlus

Effects of transient SERCA2a expression on selected transcript levels in vitro.Isolated adult ventricular myocytes were infected with Ad.SERCA2a (MOI = 100) for 48 hours, total RNA extracted and relative transcript expression analyzed in real time by qPCR using gene-specific primers. Product specificity was confirmed by post-amplification dissociation curve analysis. Ad.SERCA2a increased the expression of all genes (except MMP-7) thus confirming the effect of SERCA2a on the microarray chip. CK, creatine kinase.
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pone-0006474-g005: Effects of transient SERCA2a expression on selected transcript levels in vitro.Isolated adult ventricular myocytes were infected with Ad.SERCA2a (MOI = 100) for 48 hours, total RNA extracted and relative transcript expression analyzed in real time by qPCR using gene-specific primers. Product specificity was confirmed by post-amplification dissociation curve analysis. Ad.SERCA2a increased the expression of all genes (except MMP-7) thus confirming the effect of SERCA2a on the microarray chip. CK, creatine kinase.

Mentions: We examined whether the expression of selected transcripts was directly regulated by SERCA2a overexpression, with β-Gal serving as a control, in adult rat ventricular myocytes. Cardiomyocytes were infected with Ad.SERCA2a and Ad. β-Gal for 48 hours and the expression levels of Akt, PI3K, GLUT4, S100a3, MMP7, and CK were determined by qRT-PCR (Fig. 5). The analysis showed that SERCA2a significantly induced the expression of all molecules examined except MMP-7, which is increased in diabetic cardiomyocytes and down-regulated by SERCA2a to normal levels. Normal non-diabetic cardiomyocytes express low levels of MMP-7 which are not changed by SERCA2a.


Gene remodeling in type 2 diabetic cardiomyopathy and its phenotypic rescue with SERCA2a.

Karakikes I, Kim M, Hadri L, Sakata S, Sun Y, Zhang W, Chemaly ER, Hajjar RJ, Lebeche D - PLoS ONE (2009)

Effects of transient SERCA2a expression on selected transcript levels in vitro.Isolated adult ventricular myocytes were infected with Ad.SERCA2a (MOI = 100) for 48 hours, total RNA extracted and relative transcript expression analyzed in real time by qPCR using gene-specific primers. Product specificity was confirmed by post-amplification dissociation curve analysis. Ad.SERCA2a increased the expression of all genes (except MMP-7) thus confirming the effect of SERCA2a on the microarray chip. CK, creatine kinase.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2714457&req=5

pone-0006474-g005: Effects of transient SERCA2a expression on selected transcript levels in vitro.Isolated adult ventricular myocytes were infected with Ad.SERCA2a (MOI = 100) for 48 hours, total RNA extracted and relative transcript expression analyzed in real time by qPCR using gene-specific primers. Product specificity was confirmed by post-amplification dissociation curve analysis. Ad.SERCA2a increased the expression of all genes (except MMP-7) thus confirming the effect of SERCA2a on the microarray chip. CK, creatine kinase.
Mentions: We examined whether the expression of selected transcripts was directly regulated by SERCA2a overexpression, with β-Gal serving as a control, in adult rat ventricular myocytes. Cardiomyocytes were infected with Ad.SERCA2a and Ad. β-Gal for 48 hours and the expression levels of Akt, PI3K, GLUT4, S100a3, MMP7, and CK were determined by qRT-PCR (Fig. 5). The analysis showed that SERCA2a significantly induced the expression of all molecules examined except MMP-7, which is increased in diabetic cardiomyocytes and down-regulated by SERCA2a to normal levels. Normal non-diabetic cardiomyocytes express low levels of MMP-7 which are not changed by SERCA2a.

Bottom Line: Diabetes-associated myocardial dysfunction results in altered gene expression in the heart.Diabetic cardiomyopathic hearts have reduced levels of SERCA2a.In isolated cardiomyocytes in vitro, SERCA2a overexpression significantly modified the expression of a number of transcripts known to be involved in insulin signaling, glucose metabolism and cardiac remodeling.

View Article: PubMed Central - PubMed

Affiliation: Cardiovascular Research Center, Mount Sinai School of Medicine, New York, New York, United States of America.

ABSTRACT

Background/aim: Diabetes-associated myocardial dysfunction results in altered gene expression in the heart. We aimed to investigate the changes in gene expression profiles accompanying diabetes-induced cardiomyopathy and its phenotypic rescue by restoration of SERCA2a expression.

Methods/results: Using the Otsuka Long-Evans Tokushima Fatty rat model of type 2 diabetes and the Agilent rat microarray chip, we analyzed gene expression by comparing differential transcriptional changes in age-matched control versus diabetic hearts and diabetic hearts that received gene transfer of SERCA2a. Microarray expression profiles of selected genes were verified with real-time qPCR and immunoblotting. Our analysis indicates that diabetic cardiomyopathy is associated with a downregulation of transcripts. Diabetic cardiomyopathic hearts have reduced levels of SERCA2a. SERCA2a gene transfer in these hearts reduced diabetes-associated hypertrophy, and differentially modulated the expression of 76 genes and reversed the transcriptional profile induced by diabetes. In isolated cardiomyocytes in vitro, SERCA2a overexpression significantly modified the expression of a number of transcripts known to be involved in insulin signaling, glucose metabolism and cardiac remodeling.

Conclusion: This investigation provided insight into the pathophysiology of cardiac remodeling and the potential role of SERCA2a normalization in multiple pathways in diabetic cardiomyopathy.

Show MeSH
Related in: MedlinePlus