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: 10604334
• 负责人: BIN ZHOU
• 金额: $ 61.58万
• 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/10604334
• 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; Learning; Left; Ligation; Molecular; Molecular Biology; Morbidity - disease rate; Mus; Myocardial; Natural regeneration; Neonatal; Patients; Phenotype; Population; Proliferating; RE1-silencing transcription factor; Reporter; Repression; Research; Research Project Grants; Resources; Rest; Role; Site-Directed Mutagenesis; Systems Biology; Testing; Therapeutic; Up-Regulation; Work; cardiac regeneration; cardiac repair; cardiogenesis; cdc Genes; cell type; cofactor; combat; coronary fibrosis; fetal; gain of function; 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; preadolescence; prevent; programs; proliferation potential; recruit; repaired; single-cell RNA sequencing; therapeutic target; 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.

在该研究项目中，我们建议研究RE1沉默转录的分子机制 因子（休息）使用小鼠遗传学的协同方法调节心肌细胞细胞周期 和系统生物学。我们的最终目标是确定潜在的治疗靶标，以应对心脏病 促进心肌细胞增殖以修复受伤的成人心脏。成人心脏的维修有限 容量，因为大多数成年心肌细胞是非分散细胞。用于成人心脏的再生 现有的心肌细胞，我们需要了解如何控制心肌细胞增殖，哪个人群 心肌细胞的增殖潜力，是否具有心肌细胞的发育机制 在患病的情况下重新激活增殖，如果可以进一步执行它们。我们计划在这里 通过调查小鼠心脏中的休息功能来解决这些问题，因为我们最近的研究已经 将休息确定为胚胎心脏所需的心肌细胞增殖的新的内在调节剂 发育和新生儿心脏再生。 REST抑制细胞周期抑制剂P21维持 心肌细胞循环。表达几个关键细胞周期激活剂也需要休息。基于 根据这些发现，我们假设静止的上调可以改善受伤心脏的更新 调节细胞周期基因和调节剂的表达以促进心肌细胞增殖。我们将 在两个目标中检验该假设。 AIM 1将确定是否在失败的心中强迫休息表达 促进心肌细胞周期并改善疾病结果。我们将特别使休息在 有或没有p21失活以及强迫休息表达的心肌细胞以确定 成人心脏修复是否需要休息。 AIM 2将研究休息如何控制心肌细胞细胞 通过研究和比较成熟的剩余调节网络，在正常和失败的心脏中循环 （MYH6+）和未成熟（MYH7+）心肌细胞。我们还将通过 确定不同染色质co因子的休息募集以进行基因激活或抑制。这会 可以通过使用细胞类型特异性基因组宽息染色质免疫沉淀测序来实现 （chip-seq）和转录组学（RNA-seq），用于MYH6+与MYH7+心肌细胞的正常或受伤的心肌细胞 心。小鼠发现的人类相关性将通过关键休息的功能研究来确定 源自人诱导多能干细胞的心肌细胞增殖中的调节基因 （IPSC）。通过完成该项目，我们将确定促进心肌细胞的剩余作用 扩散以改善失败成人心脏的更新和功能。我们还将学会一个遗传 REST调节成人心肌细胞周期的调节网络，并确定了新的目标 改善心脏再生的心肌细胞增殖。新信息将推动我们的 了解心脏生物学，并为治疗心力衰竭提供新的方向。

项目成果

A sandwiched ventricular explant assay to model mouse coronary angiogenesis ex vivo.

• DOI: 10.1016/j.xpro.2023.102619
• 发表时间: 2023-12-15
• 期刊: STAR PROTOCOLS
• 作者: Lu, Pengfei;Wu, Bingruo;Wang, Yidong;Zhang, Jingran;Zhou, Bin
• 通讯作者: Zhou, Bin

Cardiac Myosin Heavy Chain Reporter Mice to Study Heart Development and Disease.

• DOI: 10.1161/circresaha.122.321461
• 发表时间: 2022-08-05
• 期刊: CIRCULATION RESEARCH
• 影响因子: 20.1
• 作者: Lu, Pengfei;Wu, Bingruo;Feng, Xuhui;Cheng, Wei;Kitsis, Richard N.;Zhou, Bin
• 通讯作者: Zhou, Bin

Prerequisite endocardial-mesenchymal transition for murine cardiac trabecular angiogenesis.

• DOI: 10.1016/j.devcel.2023.03.009
• 发表时间: 2023-05-08
• 期刊: DEVELOPMENTAL CELL
• 影响因子: 11.8
• 作者: Lu, Pengfei;Wu, Bingruo;Wang, Yidong;Russell, Megan;Liu, Yang;Bernard, Daniel J.;Zheng, Deyou;Zhou, Bin
• 通讯作者: Zhou, Bin

β-Catenin regulates endocardial cushion growth by suppressing p21.

• DOI: 10.26508/lsa.202302163
• 发表时间: 2023-09
• 期刊: Life science alliance
• 影响因子: 4.4