DNA methylation in an engineered heart tissue model of cardiac hypertrophy: common signatures and effects of DNA methylation inhibitors

Justus Stenzig, Marc N. Hirt, Alexandra Loeser, Lena M. Bartholdt, Jan-Tobias Hensel, Tessa R. Werner, Mona Riemenschneider, Daniela Indenbirken, Thomas Guenther, Christian P. Mueller, Norbert Huebner, Monika Stoll, Thomas Eschenhagen*

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

18 Citations (Web of Science)

Abstract

DNA methylation affects transcriptional regulation and constitutes a drug target in cancer biology. In cardiac hypertrophy, DNA methylation may control the fetal gene program. We therefore investigated DNA methylation signatures and their dynamics in an in vitro model of cardiac hypertrophy based on engineered heart tissue (EHT). We exposed EHTs from neonatal rat cardiomyocytes to a 12-fold increased afterload (AE) or to phenylephrine (PE 20 ?M) and compared DNA methylation signatures to control EHT by pull-down assay and DNA methylation microarray. A 7-day intervention sufficed to induce contractile dysfunction and significantly decrease promoter methylation of hypertrophy-associated upregulated genes such as Nppa (encoding ANP) and Acta1 (?-skeletal actin) in both intervention groups. To evaluate whether pathological consequences of AE are affected by inhibiting de novo DNA methylation we applied AE in the absence and presence of DNA methyltransferase (DNMT) inhibitors: 5-aza-2'-deoxycytidine (aza, 100 ?M, nucleosidic inhibitor), RG108 (60 ?M, non-nucleosidic) or methylene disalicylic acid (MDSA, 25 ?M, non-nucleosidic). Aza had no effect on EHT function, but RG108 and MDSA partially prevented the detrimental consequences of AE on force, contraction and relaxation velocity. RG108 reduced AE-induced Atp2a2 (SERCA2a) promoter methylation. The results provide evidence for dynamic DNA methylation in cardiac hypertrophy and warrant further investigation of the potential of DNA methylation in the treatment of cardiac hypertrophy.
Original languageEnglish
Article number9
JournalBasic Research in Cardiology
Volume111
Issue number1
DOIs
Publication statusPublished - Jan 2016

Keywords

  • DNA methylation
  • Tissue engineering
  • Cardiac hypertrophy
  • DNA methyl transferase
  • Heart
  • Epigenetics

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