Chromatin organization dynamics during maturation and pathological hypertrophy of cardiac myocytes

全基因组表观遗传学研究强调了心肌细胞成熟和疾病中表观遗传学过程的动力学。值得注意的是，主要的变化发生在具有顺式调控特性的基因间非编码区。染色质的三维组织使这些区域能够与远端调控区域如启动子相互作用。这些相互作用发生在高阶染色质结构中。到目前为止，还不清楚心肌细胞中发生了哪些染色质结构和相互作用，以及它们如何影响基因表达。本项目的目的是揭示小鼠心肌细胞在胎儿和出生后成熟和心力衰竭过程中染色质结构的动力学以及启动子区域的相互作用。此外，我们将证明启动子相互作用的功能相关性。为了实现这一目标，这些项目包括三个具体目标。目的A是利用高通量染色体构象捕获分析（Hi-C）阐明胎儿（E14.5）、出生后（P1）、成人和衰竭心肌细胞的染色质结构。目的B是对2500多个差异调控基因（启动子- chi - c）进行高分辨率相互作用分析，以确定相互作用的顺式调控区域。我们将整合全面的表观遗传学和基因表达数据来分析染色质结构（目的A）和相互作用（目的B）。目的C是利用CRISPR/ cas技术在es细胞源性心肌细胞中消融顺式调控区域，以证明其调控作用。该项目将首次为心肌细胞成熟和疾病期间染色质结构和启动子相互作用与表观遗传机制和基因表达的相互作用提供全基因组的见解。确定的调控机制将为新的表观遗传治疗概念的发展提供基础。

Genome-wide epigenetic studies highlight the dynamics of epigenetic processes in cardiac myocyte maturation and disease. Remarkably, major changes occur in intergenic, non-coding regions with cis-regulatory properties. The three dimensional organization of chromatin enables interactions of these regions with distal regulatory regions like i.e. promoters. These interactions occur within higher-order chromatin structures. So far, it remains unclear which chromatin structures and interactions occur in cardiac myocytes and how they influence gene expression. Aim of this project is to unravel the dynamics of chromatin structure as well as interactions of promotor regions in mouse cardiac myocytes during fetal and postnatal maturation and heart failure. In addition, we will prove the functional relevance of promotor interactions. To reach this aim, the projects consist of three specific aims. Aim A is to elucidate the chromatin structure of fetal (E14.5), postnatal (P1), adult and failing cardiac myocytes using high-throughput chromosome conformation capture analysis (Hi-C). Aim B is to perform high resolution interaction analysis for more than 2500 differentially regulated genes (promotor-Chi-C), to identify interacting cis-regulatory regions. We will integrate comprehensive epigenetic and gene expression data in the analysis of chromatin structure (aim A) and interactions (aim B). Aim C is to ablate cis-regulatory regions using the CRISPR/Cas-Technology in ES-cell derived cardiac myocytes, to prove their regulatory role.For the first time, this project will provide genome-wide insights into the interplay of chromatin structure and promotor interactions with epigenetic mechanisms and gene expression in cardiac myocytes during maturation and in disease. The identified regulatory mechanisms will provide a basis for the development of novel epigenetic therapeutic concepts.