DNA methylation in an engineered heart tissue model of cardiac hypertrophy: common signatures and effects of DNA methylation inhibitors
DNA methylation in an engineered heart tissue model of cardiac hypertrophy: common signatures and effects of DNA methylation inhibitors
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DOI：
10.1007/s00395-015-0528-z
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发表时间：
2016-01-01
影响因子：
9.5
通讯作者：
Eschenhagen, Thomas
中科院分区：
文献类型：
--
作者：
Stenzig, Justus;Hirt, Marc N.;Eschenhagen, Thomas
DNA甲基化影响转录调控，并构成癌症生物学中的药物靶标。在心肌肥大中，DNA甲基化可能控制胎儿基因程序。因此，我们研究了DNA甲基化的签名和他们的动力学在体外模型的基础上，心脏肥大的工程心脏组织（EHT）。我们暴露EHTs从新生大鼠心肌细胞增加12倍后负荷（AE）或苯丙氨酸（PE 20 μ M），并比较DNA甲基化的签名，以控制EHT下拉分析和DNA甲基化微阵列。在两个干预组中，7天的干预足以诱导收缩功能障碍并显著降低肥大相关上调基因（如Nppa（编码ANP）和Acta 1（α-骨骼肌动蛋白））的启动子甲基化。为了评价AE的病理学后果是否受到抑制从头DNA甲基化的影响，我们在不存在和存在DNA甲基转移酶（DNMT）抑制剂的情况下应用AE：5-氮杂-2 '-脱氧胞苷（aza，100 μ M，核苷抑制剂）、RG 108（60 μ M，非核苷）或亚甲基二异辛酸（MDSA，25 μ M，非核苷）。Aza对EHT功能没有影响，但RG 108和MDSA部分防止了AE对力、收缩和舒张速度的不利影响。RG 108降低AE诱导的Atp 2a 2（SERCA 2a）启动子甲基化。这些结果为心肌肥厚中的动态DNA甲基化提供了证据，并保证了进一步研究DNA甲基化在治疗心肌肥厚中的潜力。
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 mu 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 (alpha-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 mu M, nucleosidic inhibitor), RG108 (60 mu M, non-nucleosidic) or methylene disalicylic acid (MDSA, 25 mu 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.