Control of cardiomyocyte cell cycle by REST in heart failure and regeneration

通过 REST 控制心力衰竭和再生中的心肌细胞周期

基本信息

批准号：
10397428
负责人：
BIN ZHOU
金额：
$ 62.93万
依托单位：
ALBERT EINSTEIN COLLEGE OF MEDICINE
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-07-15 至 2024-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10397428
关键词：
Address Adult Anterior Arteries Biology Candidate Disease Gene Cardiac Cardiac Myocytes Cell Cycle Cells ChIP-seq Chromatin Cytokinesis Data Development Disease Disease Outcome Embryo Embryonic Heart Epigenetic Process Gene Activation Genes Genetic Goals Heart Heart Diseases Heart Injuries Heart failure Human Inherited Interphase Cell Knock-out Knowledge Label Left Ligation Molecular Molecular Biology Morbidity - disease rate Mus Myocardial Natural regeneration Neonatal Patients Phenotype Population RE1-silencing transcription factor Reporter Repression Research Research Project Grants Resources Rest Role Site-Directed Mutagenesis Systems Biology Testing Therapeutic Up-Regulation Work base cardiac regeneration cardiac repair cardiogenesis cdc Genes cell type combat coronary fibrosis fetal fetal reactivity gene network gene repression genome-wide heart function histone modification improved improved outcome induced pluripotent stem cell induced pluripotent stem cell derived cardiomyocytes inhibitor injured injury and repair innovation insight loss of function mortality mouse genetics mouse model neovascularization novel novel therapeutics overexpression prevent programs recruit regenerative biology repaired single-cell RNA sequencing therapeutic target transcriptome sequencing transcriptomics

项目摘要

In this research project, we propose to study the molecular mechanisms by which RE1 silencing transcription factor (REST) regulates cardiomyocyte cell cycle using synergistic approaches of mouse genetics, molecular and systems biology. Our ultimate goal is to identify potential therapeutic targets to combat heart disease by promoting cardiomyocyte proliferation to repair the injured adult heart. The adult heart has a limited repairing capacity, because most adult cardiomyocytes are non-dividing cells. For adult heart regeneration by pre- existing cardiomyocytes, we need to understand how cardiomyocyte proliferation is controlled, what population of cardiomyocytes maintains proliferation potential, whether developmental mechanisms of cardiomyocyte proliferation are re-activated under diseased conditions, and if they can be further enforced. Here we plan to address these questions by investigating REST functions in the mouse heart, because our recent studies have identified REST as a new intrinsic regulator of cardiomyocyte proliferation required for embryonic heart development and neonatal heart regeneration. REST represses the cell cycle inhibitor p21 to maintain cardiomyocyte cell cycling. REST is also required for the expression of several key cell cycle activators. Based on these findings, we hypothesize that upregulation of REST may improve the renewal of the injured heart by modulating the expression of cell cycle genes and regulators to promote cardiomyocyte proliferation. We will test this hypothesis in two aims. Aim 1 will determine whether forced REST expression in failing hearts promotes cardiomyocyte cell cycle and improves the disease outcome. We will specifically inactivate REST in the cardiomyocytes, with or without the p21 inactivation, as well as forced REST expression, to determine whether REST is required for adult heart repair. Aim 2 will investigate how REST controls cardiomyocyte cell cycle in normal and failing hearts by studying and comparing the REST regulatory network in the mature (MYH6+) and immature (MYH7+) cardiomyocytes. We will also study the REST regulatory network by determining the REST recruitment of different chromatin co-factors for gene activation or repression. This will be achieved by using the cell-type specific, genome wide REST chromatin immunoprecipitation sequencing (ChIP-seq) and transcriptomics (RNA-seq) for MYH6+ versus MYH7+ cardiomyocytes from normal or injured hearts. The human relevance of mouse findings will be ascertained by the functional study of key REST- regulated genes in the proliferation of cardiomyocytes derived from the human inducible pluripotent stem cells (iPSC). By completing this project, we will have determined the REST role in promoting cardiomyocyte proliferation to improve the renewal and function of failing adult hearts. We will have also learned a genetic regulatory network by which REST regulates adult cardiomyocyte cell cycle and identified new targets for improving cardiomyocyte proliferation for heart regeneration. The new information will advance our understanding of cardiac biology and provide a new direction to treat heart failure.

在这个研究项目中，我们拟研究RE1沉默转录的分子机制因子（REST）利用小鼠遗传学、分子生物学的协同方法调节心肌细胞周期和系统生物学。我们的最终目标是通过以下方式确定对抗心脏病的潜在治疗靶点促进心肌细胞增殖以修复受损的成人心脏。成人心脏的修复能力有限能力，因为大多数成年心肌细胞是非分裂细胞。用于成人心脏再生现有的心肌细胞，我们需要了解心肌细胞增殖是如何控制的，哪些人群心肌细胞保持增殖潜力，无论心肌细胞的发育机制如何在患病条件下，如果可以进一步加强，增殖就会重新激活。在这里我们计划通过研究小鼠心脏的 REST 功能来解决这些问题，因为我们最近的研究表明确定 REST 是胚胎心脏所需的心肌细胞增殖的新内在调节因子发育和新生儿心脏再生。 REST 抑制细胞周期抑制剂 p21 以维持心肌细胞的细胞周期。几种关键细胞周期激活剂的表达也需要 REST。基于根据这些发现，我们假设 REST 的上调可能会通过以下方式改善受损心脏的更新：调节细胞周期基因和调节因子的表达，促进心肌细胞增殖。我们将在两个目标上检验这个假设。目标 1 将确定衰竭心脏中是否强制表达 REST 促进心肌细胞周期并改善疾病结果。我们将专门停用 REST 心肌细胞，有或没有 p21 失活，以及强制 REST 表达，以确定成人心脏修复是否需要休息。目标 2 将研究 REST 如何控制心肌细胞通过研究和比较成熟心脏中的 REST 调节网络，研究正常心脏和衰竭心脏的循环 (MYH6+) 和未成熟的 (MYH7+) 心肌细胞。我们还将研究 REST 监管网络确定不同染色质辅助因子的 REST 招募以激活或抑制基因。这将通过使用细胞类型特异性、全基因组 REST 染色质免疫沉淀测序来实现正常或受损的 MYH6+ 与 MYH7+ 心肌细胞的 (ChIP-seq) 和转录组学 (RNA-seq) 心。小鼠研究结果与人类的相关性将通过关键 REST 的功能研究来确定人诱导多能干细胞来源的心肌细胞增殖中的调控基因（iPSC）。通过完成这个项目，我们将确定 REST 在促进心肌细胞中的作用增殖以改善衰竭成人心脏的更新和功能。我们还将学到遗传 REST 调节成人心肌细胞周期的调节网络并确定了新的靶标改善心肌细胞增殖以促进心脏再生。新信息将推动我们了解心脏生物学并为治疗心力衰竭提供新方向。