Cardiac Myocyte De Novo DNA Methyltransferases 3a/3b Are Dispensable for Cardiac Function and Remodeling after Chronic Pressure Overload in Mice

PLoS One. 2015 Jun 22;10(6):e0131019. doi: 10.1371/journal.pone.0131019. eCollection 2015.

Abstract

Background: Recent studies reported altered DNA methylation in failing human hearts. This may suggest a role for de novo DNA methylation in the development of heart failure. Here, we tested whether cardiomyocyte-specific loss of de novo DNA methyltransferases Dnmt3a and Dnmt3b altered cardiac function and remodeling after chronic left ventricular pressure overload.

Methods: Mice with specific ablation of Dnmt3a and Dnmt3b expression in cardiomyocytes were generated by crossing floxed Dnmt3afl and Dnmt3bfl alleles with mice expressing Cre recombinase under control of the atrial myosin light chain gene promoter. The efficacy of combined Dnmt3a/3b ablation (DKO) was characterized on cardiomyocyte-specific genomic DNA and mRNA levels. Cardiac phenotyping was carried out without (sham) or with left ventricular pressure overload induced by transverse aortic constriction (TAC). Under similar conditions, cardiac genome-wide transcriptional profiling was performed and DNA methylation levels of promoters of differentially regulated genes were assessed by pyrosequencing.

Results: DKO cardiomyocytes showed virtual absence of targeted Dnmt3a and Dnmt3b mRNA transcripts. Cardiac phenotyping revealed no significant differences between DKO and control mice under sham and TAC conditions. Transcriptome analyses identified upregulation of 44 and downregulation of 9 genes in DKO as compared with control sham mice. TAC mice showed similar changes with substantial overlap of regulated genes compared to sham. Promoters of upregulated genes were largely unmethylated in DKO compared to control mice.

Conclusion: The absence of cardiac pathology in the presence of the predicted molecular phenotype suggests that de novo DNA methylation in cardiomyocytes is dispensable for adaptive mechanisms after chronic cardiac pressure overload.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • DNA (Cytosine-5-)-Methyltransferases / genetics*
  • DNA / genetics
  • DNA Methylation / genetics
  • DNA Methyltransferase 3A
  • DNA Methyltransferase 3B
  • Heart / physiology*
  • Heart Failure / genetics
  • Heart Failure / physiopathology
  • Male
  • Mice
  • Myocytes, Cardiac / enzymology*
  • Myocytes, Cardiac / physiology*
  • Pressure
  • Promoter Regions, Genetic / genetics
  • RNA, Messenger / genetics
  • Transcriptome / genetics
  • Ventricular Remodeling / genetics
  • Ventricular Remodeling / physiology*

Substances

  • DNMT3A protein, human
  • Dnmt3a protein, mouse
  • RNA, Messenger
  • DNA
  • DNA (Cytosine-5-)-Methyltransferases
  • DNA Methyltransferase 3A

Associated data

  • GEO/GSE68518

Grants and funding

This study was supported by the Deutsche Forschungsgemeinschaft, Bonn, Germany (SFB992, project B03). (www.dfg.de) The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.