Myocardial Repair with a Novel Engineered Cardiac Muscle Patch

使用新型工程心肌补片修复心肌

• 批准号: 10229464
• 负责人: Wuqiang Zhu
• 金额: $ 41.28万
• 依托单位: MAYO CLINIC ARIZONA
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-01 至 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10229464
• 关键词: Address; Animal Model; Anions; Area; Arrhythmia; Bioenergetics; CCND2 gene; Cardiac; Cardiac Myocytes; Cell Cycle; Cell Death; Cell Line; Cell Proliferation; Cell Survival; Cell Therapy; Cells; Chronic; Cicatrix; Clinical; Coupling; Data; Development; Donor person; Electric Stimulation; Electrophysiology (science); Engineering; Engraftment; Exhibits; Exposure to; Family suidae; Fibrin; Ganciclovir; Genetic Induction; Goals; Heart; Heart Injuries; Histology; Human; Incidence; Infarction; Knowledge; Light; Magnetic Resonance Imaging; Magnetic Resonance Spectroscopy; Mediating; Modeling; Monitor; Mus; Muscle Cells; Myocardial; Myocardial Infarction; Myocardial Ischemia; Myocardium; Myosin Heavy Chains; Natural regeneration; Organ; Predisposition; Protocols documentation; Publications; Regenerative research; Reporting; Rhodopsin; Risk; Rodent Model; Safety; Simplexvirus; Structure; Surface; Tachyarrhythmias; Techniques; Testing; Tetracyclines; Therapeutic; Tissue Engineering; Tissue Grafts; Tissues; Transgenes; Transgenic Organisms; Translating; Transplantation; Ventricular; Ventricular Cardiac alpha-Myosin; adverse outcome; beneficiary; cardiac regeneration; cardioprotection; clinical implementation; clinically relevant; experimental study; functional improvement; functional outcomes; functional restoration; heart function; heart rhythm; human embryonic stem cell; immunosuppressed; improved; in vivo; induced pluripotent stem cell; injured; ischemic injury; loss of function; mouse model; nonhuman primate; novel; novel strategies; overexpression; porcine model; promoter; regenerative; repaired; suicide gene; tumor

Project Summary Cellular transplantation has emerged as a promising therapeutic approach for myocardial repair. However, several critical issues remain to be addressed which include, but are not limited to: 1) low donor cell engraftment rate (ranging from 0.1-10% in previous publications); 2) lack of knowledge on the mechanisms underlying the in vivo beneficiary effects of grafted cells. Understanding the in vivo effects of grafted cells may promote the development of more effective cardioprotective strategies. Several groups reported that applying prefabricated cardiac tissue, a "cardiac muscle patch" (CMP) made of hiPSC-derived cardiac cells, effectively increased engraftment rate. We recently established a novel strategy which has been demonstrated to significantly enhance engraftment rate. Specifically, we established a human induced pluripotent stem cell (hiPSC) line which carries a transgene encoding for the human CCND2 (Cyclin D2) driven by the cardiomyocyte specific α-myosin heavy chain (α-MHC) promoter. CCND2-overexpressing hiPSC- derived cardiomyocytes (hiPSC-CCND2OECMs) exhibits increased cell cycle activity and cell proliferation compared with genetically naïve hiPSC-CMs expressing wild-type levels of CCND2 (hiPSC-CCND2WTCMs). In a mouse model of myocardial infarction (MI), the number of engrafted cells was tripled in hearts injected with hiPSC-CCND2OECMs compared to those receiving hiPSC-CCND2WTCMs 4 weeks post MI and transplantation, resulting in significantly smaller infarct size and improved cardiac function. These data suggests that transgenic CCND2 overexpression in hiPSC-CM grafts constitutes a viable approach to enhance engraftment and restore function in ischemic heart disease. The proposal will develop a novel human cardiac muscle patch with hiPSC-CCND2OECMs (designated as hCMP- CCND2OECMs), assess their capability to continuously remuscularize the injured heart in the long term and ultimately replace the scar tissue, and test their safety and translational potential in a large animal model (the pig MI model). In the case transplanted hCMPs improve cardiac function in the chronically infarcted pig hearts, the proposal also determine if this functional improvement is attributable to remuscularization or other mechanisms. Specifically, we aimed to address the yet unanswered, but profoundly important, question whether hCMPs improve cardiac function via direct donor-host coupling in pig MI model. Our long term goal is to develop a heart regeneration strategy that can be translated to humans. Two Specific Aims are proposed. Specific Aim 1 will test the hypothesis that this novel hCMP continuously remuscularize injured myocardium, replacing transmural scar tissue, and improve cardiac function in infarcted pig hearts. We will determine (i) the impact of patch transplantation on cardiac structure and function, and (ii) safety of patch transplantation (susceptibility to inducible arrhythmias, and risk of tumor formation). Specific Aim 2 test the hypothesis that the magnitude of functional changes positively correlates with the number of donor cell-derived CMs. We will determine whether (i) donor cell survival is required for sustained improvement of cardiac function, and (ii) whether improvement in cardiac function is mediated, at least in part, by electromechanical coupling of transplanted donor myocytes.

项目摘要 细胞移植已成为心肌修复的一种有前途的治疗方法。但是，在这方面， 几个关键问题仍有待解决，包括但不限于：1）低供体细胞移植 （2）缺乏对感染机制的了解;（3）缺乏对感染机制的了解;（4）缺乏对感染机制的了解。 移植细胞的体内受益效应。了解移植细胞的体内效应可能会促进 开发更有效的心脏保护策略。 几个研究小组报告说，应用预制的心脏组织，一种由 hiPSC衍生的心肌细胞，有效地增加了植入率。我们最近制定了一个新的策略 其已被证明显著提高植入率。具体来说，我们建立了一个人类 携带编码人CCND 2（细胞周期蛋白D2）的转基因的诱导多能干细胞（hiPSC）系 由心肌细胞特异性α-肌球蛋白重链（α-MHC）启动子驱动。CCND 2-过表达hiPSC- 衍生的心肌细胞（hiPSC-CCND 2 OECMs）表现出增加的细胞周期活性和细胞增殖 与表达野生型水平的CCND 2的遗传幼稚hiPSC-CM（hiPSC-CCND 2 WTCM）相比。在 一种心肌梗死（MI）的小鼠模型，在注射了 与MI和移植后4周接受hiPSC-CCND 2 WTCM的那些相比， 导致显著更小的梗塞面积和改善的心脏功能。这些数据表明， hiPSC-CM移植物中的CCND 2过表达构成了增强植入和恢复的可行方法。 在缺血性心脏病中的作用 该提案将开发具有hiPSC-CCND 2 OECMs（命名为hCMP-2 OECMs）的新型人类心肌贴片。 CCND 2 OECMs），评估其长期持续重建受损心脏肌肉的能力， 最终替代疤痕组织，并在大型动物模型中测试其安全性和转化潜力（ 猪MI模型）。在移植的hCMP改善慢性梗塞猪心脏的心脏功能的情况下， 该建议还确定这种功能改善是否可归因于肌肉再生或其他 机制等具体来说，我们的目标是解决尚未回答的，但非常重要的问题，hCMP是否 在猪MI模型中通过直接供体-宿主偶联改善心脏功能。我们的长期目标是培养一颗 可以转化为人类的再生策略。提出了两个具体目标。 特异性目的1将检验这种新型hCMP持续地使受损心肌肌化的假设， 替换透壁瘢痕组织，并改善梗塞猪心脏的心脏功能。我们将确定（i） 补片移植对心脏结构和功能影响及补片移植的安全性 （对可诱导心律失常的易感性和肿瘤形成的风险）。 具体目标2检验功能变化的幅度与数量正相关的假设。 供体细胞来源的CM。我们将确定（i）持续改善是否需要供体细胞存活 以及（ii）心脏功能的改善是否至少部分由以下因素介导： 移植供体肌细胞的机电耦合。