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.

项目概要 我们小组最近的研究表明，人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。 20 个射血分数点与强健的肌肉重建相关，通常会产生厘米级的移植物 然而，随着临床试验的进展，仍然存在一些悬而未决的问题。 hESC-CM的作用机制为何相对较低？ 如果我们添加关键的心肌细胞类型，植入会得到改善吗？ 除了心肌细胞之外？在目标 1 中，我们讨论了 hESC-CM 的作用机制是否与以下因素有关： 直接细胞替换，或者是否存在显着的旁分泌成分 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，产生完整的非收缩心肌细胞 将比较这些“仅旁分泌”心肌细胞的肌原纤维、动作电位和钙瞬变。 在目标 2 中，我们将测试野生型细胞修复梗塞大鼠心脏的假设。 “智能水凝胶”旨在通过 Notch 通路发出信号，可以通过 hiPSC 改善心脏再生 这种 Notch 凝胶可在植入后刺激 hiPSC-CM 增殖并促进血管向内生长。 我们将测试这些结构性益处是否伴随着周围宿主微循环。 最后，目标 3 是我们最近完成的大鼠研究的后续研究。 观察到 hESC 衍生的心外膜细胞 (hESC-Epi) 与 hESC-CM 在促进 我们将利用我们的猕猴来增强梗塞心脏的再肌肉化和功能恢复。 猴子模型测试 hESC-Epi 是否增强基于 hESC-CM 的心脏再生，假设 这些细胞将促进 hESC-CM 成熟并增强其增殖，从而减少致心律失常 该提案中的研究将直接影响我们的研究。 即将进行的心脏修复临床试验。