Metabolic and Transcriptional Reprogramming of Cardiac Maturation

心脏成熟的代谢和转录重编程

基本信息

批准号：
10202988
负责人：
Charles E Murry
金额：
$ 61.77万
依托单位：
UNIVERSITY OF WASHINGTON
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-04-01 至 2025-03-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10202988
关键词：
Adult Animal Model Animal Testing Appearance Arrhythmia Attenuated Automobile Driving Awareness Calcium Carbohydrates Cardiac Cardiac Myocytes Cell Cycle Cell Nucleus Cells Cellular Metabolic Process Clinical Clinical Trials Clustered Regularly Interspaced Short Palindromic Repeats Communities Complex Data Databases Disease model Electrophysiology (science)Elements Energy Metabolism Engraftment Enzymes Epigenetic Process Exhibits Family suidae Fatty Acids Gene Expression Gene Expression Profile Gene Expression Regulation Genes Genetic Transcription Glucocorticoids Goals Heart Heart Diseases Heart Injuries Human Hypertrophy In Situ In Vitro Individual Intervention Ion Channel Lead Left Macaca Metabolic Metabolic hormone Metabolism Miniature Swine Monkeys Muscle Myocardial Natural regeneration Nuclear Patients Physiological Pluripotent Stem Cells Process Production Protocols documentation Pump Rattus Regulation Resources Science Series Severities Stimulus Structure Techniques Telemetry Testing Text Time Transplantation Ventricular Arrhythmia base cardiac regeneration cardiac repair clinically relevant combinatorial drug discovery feeding gain of function heart function heart rhythm human pluripotent stem cell immunosuppressed in vivo in vivo evaluation metabolome metabolomics multi-electrode arrays nonhuman primate overexpression patch clamp porcine model programs regenerative therapy restoration single-cell RNA sequencing stem cells success trait transcription factor transcriptional reprogramming transcriptome sequencing

项目摘要

Project Summary Recent advances in stem cell science have led to accelerated progress in cardiac regeneration. Our group successfully achieved large-scale re-muscularization of the infarcted hearts of macaque monkeys by transplanting human cardiomyocytes derived from pluripotent stem cells (hPSC-CMs). These cardiomyocytes restored ejection from ~40% to ~62%, the largest restoration of cardiac function of which we are aware. This therapy is complicated by the appearance of transient ventricular arrhythmias, which last for several weeks before disappearing. Our electrophysiological studies indicate that the arrhythmias result from pacemaking activity, which in turn results from the immaturity of the hPSC-CMs at the time of transplantation. The main goal of this proposal is to enhance the maturation of hPSC-CM to make them non-arrhythmogenic, using metabolic and transcriptional reprogramming. In Aim 1 we build on our observations that modulating metabolism has wide-ranging effects on maturation, including reducing automaticity, and increasing physiological hypertrophy, force production, and more adult-like calcium cycling. The most powerful metabolic interventions, substrate switching, metabolic hormones, and energy sensing, will be systematically optimized to enhance electrical maturity in vitro. Once optimized, we will explore the underlying mechanisms, and then test whether this maturation reduces arrhythmias by transplanting them into porcine hearts. Aim 2 takes advantage of a recently generated resource, where we performed RNA-seq on a timed series of human myocardial grafts in the rat heart as they matured to adult levels in vivo. By comparing these data to immature cardiomyocytes, we identified a set of transcriptional regulators that are candidate drivers of maturation. We will perform CRISPR-based gain-of-function studies to activate these factors in vitro. Using gene expression and electrophysiology analyses, we will then identify optimal combinations to enhance maturation. If successful, these studies will solve the greatest barrier to stem cell-based heart regeneration and bring us much closer to clinical trials.

项目概要干细胞科学的最新进展加速了心脏再生的进展。我们组成功实现了猕猴梗塞心脏的大规模再肌化移植源自多能干细胞（hPSC-CM）的人类心肌细胞。这些心肌细胞射血量从约 40% 恢复至约 62%，这是我们所知的最大的心功能恢复。这持续数周的短暂性室性心律失常的出现使治疗变得复杂消失之前。我们的电生理学研究表明心律失常是由起搏引起的活性，这又是移植时 hPSC-CM 不成熟的结果。主要该提案的目标是促进 hPSC-CM 的成熟，使其不致心律失常，使用代谢和转录重编程。在目标 1 中，我们以我们的观察为基础，调节新陈代谢对成熟具有广泛的影响，包括降低自动性和增加生理肥大、力量产生以及更像成人的钙循环。最强大的新陈代谢干预措施、底物转换、代谢激素和能量感应将得到系统优化，以增强体外电成熟度。优化完成后，我们将探索底层机制，然后进行测试这种成熟是否可以通过将其移植到猪心脏中来减少心律失常。目标2利用最近生成的资源，我们对一系列定时的人类心肌移植物进行了 RNA 测序当它们在体内成熟到成年水平时，在大鼠心脏中。通过将这些数据与未成熟的心肌细胞进行比较，我们确定了一组转录调节因子，它们是成熟的候选驱动因素。我们将表演基于 CRISPR 的功能获得研究可在体外激活这些因子。利用基因表达和通过电生理学分析，我们将确定促进成熟的最佳组合。如果成功的话，这些研究将解决基于干细胞的心脏再生的最大障碍，并使我们更接近于临床试验。