Subtype Spec.& Arrhythmogenica Potential of Stem Cell Derived Cardiomyocytes

子类型规格

• 批准号: 8676868
• 负责人: MICHAEL Alan LAFLAMME
• 金额: $ 29.07万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至 2017-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8676868
• 关键词: Address; Agonist; Animals; Arrhythmia; Behavior; Biological Models; Biological Pacemakers; Cardiac; Cardiac Myocytes; Cardiovascular system; Cell Culture Techniques; Cell Transplantation; Cells; Coupled; Coupling; Data; Disease; Electrocardiogram; Elements; Event; Genes; Genetic; Goals; Heart; Heart Atrium; Heart Transplantation; Image; Incidence; Infarction; Label; Lead; Mediating; Methods; Modeling; Molecular; Monitor; Mus; Muscle; Myocardial Infarction; Myocardium; NRG1 gene; Neuregulins; Nodal; Oryctolagus cuniculus; Pacemakers; Phenotype; Play; Population; Property; Proteins; Rattus; Receptor Cell; Reporter; Reporting; Role; Sick Sinus Syndrome; Signal Transduction; Stem cells; System; Testing; Transplantation; Uncertainty; Ventricular; Ventricular Arrhythmia; Work; calcium indicator; cardiac repair; cell preparation; cell type; electrical property; genetic selection; human embryonic stem cell; in vivo; nodal myocyte; novel; promoter; repaired; response

Project 2: Subtype Specification and Arrhythmogenic Potential of Stem Cell Derived Cardiomyocytes. Cardiomyocytes derived from human embryonic stem cells (hESCs) have tremendous promise for cardiac repair, both for replacing ventricular myocardium after an infarct and serving as a "biologic pacemaker" in diseases like sick-sinus syndrome. We have reported methods to generate large quantities of highly purified cardiomyocytes from hESCs, but these preparations include cells with distinct nodal/pacemaker and "working" (i.e. chamber-like) phenotypes. For these cells to be useful and safe, they must have electrophysiological properties matched to each application. Hence, this project will develop approaches to control the cardiac subtype of hESC derivatives and then will determine their electrophysiological behavior following transplantation in intact and infarcted hearts. In Aim 1, we will develop a genetic selection system to follow the lineage relationships of cardiac subtypes derived from hESCs. This work builds on our preliminary findings that a promoter element from the cGATA6 gene identifies hESC-derived cardiomyocytes with the nodal phenotype. In Aim 2, we will elucidate the molecular events by which neuregulin/ErbB signaling regulates the cardiac subtype of hESC-derived cardion\yocytes. This work will employ the genetic reporters from Aim 1 but will also lead to complementary pharmacological approaches to control cardiac subtype specification. In Aim 3, we will address two uncertainties regarding the electrophysiological behavior of hESC-derived cardiomyocytes following transplantation. First, we will test whether these cells are electrically coupled and beat synchronously with host muscle in normal and infarcted hearts. Second, we will test the hypothesis that their transplantation will modulate the incidence of infarct-related arrhythmias, while the incidence of graft-associated arrhythmias can be "tuned" by controlling the phenotype of the input cell preparation.

项目 2：干细胞来源的心肌细胞的亚型规范和致心律失常潜力。 源自人胚胎干细胞 (hESC) 的心肌细胞在心脏领域具有巨大的前景 修复，既可用于更换梗死后的心室心肌，又可作为“生物起搏器” 病态窦房结综合征等疾病。我们已经报道了产生大量高度 从 hESC 纯化心肌细胞，但这些制剂包括具有不同淋巴结/起搏器的细胞 和“工作”（即类似室）表型。为了使这些细胞有用且安全，它们必须具有 与每个应用相匹配的电生理特性。因此，该项目将开发方法 控制hESC衍生物的心脏亚型，进而决定其电生理行为 在完整心脏和梗塞心脏移植后。在目标 1 中，我们将开发遗传选择系统 追踪源自 hESC 的心脏亚型的谱系关系。这项工作建立在我们的 初步发现，来自 cGATA6 基因的启动子元件可识别 hESC 来源 具有结节表型的心肌细胞。在目标 2 中，我们将阐明分子事件 神经调节蛋白/ErbB 信号传导调节 hESC 衍生的心肌细胞的心脏亚型。这项工作将采用目标 1 中的遗传报告基因，但也将导致控制心脏亚型规范的补充药理学方法。在目标 3 中，我们将解决移植后 hESC 衍生心肌细胞电生理行为的两个不确定性。首先，我们将测试这些细胞是否在正常和正常状态下与宿主肌肉电耦合并同步跳动。 梗塞的心脏。其次，我们将测试以下假设：它们的移植将调节梗塞相关心律失常的发生率，而移植物相关心律失常的发生率可以通过控制输入细胞制剂的表型来“调节”。