Mechanisms of Cell-Based Heart Regeneration

基于细胞的心脏再生机制

• 批准号: 10371893
• 负责人: Charles E Murry
• 金额: $ 87.12万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-04-01 至 2025-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10371893
• 关键词: Action Potentials; Acute; Address; Animals; Area; Arrhythmia; Attenuated; Automobile Driving; Biotechnology; Blood Vessels; CRISPR/Cas technology; Calcium; Cardiac; Cardiac Myocytes; Cell Therapy; Cell Transplantation; Cells; Chronic; Clinical Trials; Clustered Regularly Interspaced Short Palindromic Repeats; Collagen; Complex; Contracts; EFRAC; Engineering; Engraftment; Fibroblasts; Gel; Genes; Guinea; Heart; Heart Diseases; Human; Hydrogels; Immobilization; Immune response; Infarction; Inferior; Knock-out; Knowledge; Ligands; Macaca; Macaca mulatta; Magnetic Resonance Imaging; Mechanics; Microcirculation; Modeling; Monkeys; Mus; Myocardial; Myocardial Infarction; Myocardium; Myofibrils; Natural regeneration; Operative Surgical Procedures; Paracrine Communication; Pathway interactions; Patients; Pluripotent Stem Cells; Population; Primates; Protocols documentation; Rattus; Recovery; Recovery of Function; Research; Role; Signal Transduction; Smooth Muscle Myocytes; System; Telemetry; Testing; Time; Tissue Engineering; Tissues; Transplantation; Troponin; Vascularization; Ventricular Arrhythmia; Ventricular Function; Work; base; cardiac implant; cardiac regeneration; cardiac repair; cardiac tissue engineering; cell type; design; heart function; human embryonic stem cell; human embryonic stem cell transplantation; improved; improved functioning; nonhuman primate; notch protein; paracrine; prevent; progenitor; repaired; skeletal; therapeutically effective; treatment optimization

Project Summary Recent studies from our group demonstrate that human embryonic stem cell-derived cardiomyocytes (hESC- CM) can improve the function of infarcted hearts of macaque monkeys. These improvements of as much as 20 ejection fraction points are associated with robust remuscularization, often giving centimeter-scale grafts that are visible by MRI. As we progress toward clinical trials, however, several outstanding questions remain unanswered. What is the mechanism of hESC-CM action? How can the relatively low efficiency of cardiac engraftment be improved? Will we get more complete regeneration if we add key myocardial cell types in addition to cardiomyocytes? In Aim 1 we address whether the mechanism of hESC-CM action is related to direct cell replacement, or if there is a significant paracrine component. To test this, we have used CRISPR- Cas9 to delete cardiac and skeletal TNNI genes in hiPSCs, yielding non-contractile cardiomyocytes with intact myofibrils, action potentials and calcium transients. These “paracrine-only” cardiomyocytes will be compared to wild type cells for their ability to repair the infarcted rat heart. In Aim 2 we will test the hypothesis that a “smart hydrogel”, designed to signal through the Notch pathway, can improve cardiac regeneration with hiPSC- CMs. This Notch gel stimulates hiPSC-CM proliferation after engraftment and promotes vascular ingrowth from the surrounding host microcirculation. We will test if these structural benefits are accompanied by enhanced ventricular function. Finally, Aim 3 follows up on recently completed studies in the rat, where we observed that hESC-derived epicardial cells (hESC-Epi) are synergistic with hESC-CMs in terms of promoting enhanced remuscularization and functional recovery of the infarcted heart. We will utilize our macaque monkey model to test whether hESC-Epi augment hESC-CM-based heart regeneration, with the hypothesis that these cells will promote hESC-CM maturation and enhance their proliferation, resulting in less arrhythmogenic grafts that more completely remuscularize the infarct. Studies in this proposal will impact directly on our upcoming clinical trials of cardiac repair.

项目摘要 我们小组的最新研究表明，人类胚胎干细胞衍生的心肌细胞（HESC- CM）可以改善猕猴梗塞心脏的功能。这些改进和 20个射血分数与强大的回复相关，通常给予厘米尺度的移植物 MRI可见。然而，随着我们朝着临床试验的进展，仍然存在几个出色的问题 未得到答复。 HESC-CM动作的机制是什么？心脏相对低效率如何 改善植入？如果我们在中添加关键心肌细胞类型，我们会获得更完整的再生吗 增加心肌细胞？在AIM 1中，我们解决了hESC-CM动作的机制是否与 直接替代细胞，或者是否存在明显的旁分泌成分。为了测试这一点，我们使用了CRISPR- Cas9删除HIPSC中的心脏和骨骼TNNI基因，产生完整的非合同心肌细胞 肌原纤维，作用电位和钙瞬变。将比较这些“仅旁分泌”心肌细胞 到野生型细胞修复梗塞大鼠心脏的能力。在AIM 2中，我们将检验以下假设。 “智能水凝胶”旨在通过Notch途径发出信号，可以通过HIPSC-改善心脏再生。 CMS。这种缺口凝胶刺激植入后HIPSC-CM增殖并促进血管内部 从周围的主体微循环中。我们将测试这些结构性益处是否伴随 增强的心室功能。最后，AIM 3跟进了最近在老鼠中完成的研究，我们在这里 观察到hESC衍生的心外膜细胞（HESC-EPI）在促进方面与HESC-CMS协同作用 梗塞心脏的重新震动和功能恢复增强。我们将利用猕猴 猴子模型测试HESC-EPI是否增加了基于HESC-CM的心脏再生，并具有假设 这些细胞将促进hESC-CM成熟并增强其增殖，从而降低心律失常 移植更完全震撼了梗塞。该提案中的研究将直接影响我们 即将进行的心脏修复临床试验。