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

• 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
• 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

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 通过其调节成体心肌细胞周期并确定了新靶标 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。