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有前途的治疗方法。然而， 仍有几个关键问题要解决，其中包括但不限于：1）低供体细胞植入 利率（以前出版物的0.1-10％范围）； 2）缺乏对IN的机制的知识 接枝细胞的体内受益人作用。了解接枝细胞的体内影响可能会促进 制定更有效的心脏保护策略。 几个小组报告说，采用预制心脏组织，一种由制成的“心肌斑块”（CMP） HIPSC衍生的心细胞有效提高了参与率。我们最近建立了一种新颖的策略 已证明可以显着提高植入率。具体来说，我们建立了一个人 诱导多能干细胞（HIPSC）系列，该系带有编码人CCND2的转基因（Cyclin d2） 由心肌细胞特异性α-肌球蛋白重链（α-MHC）启动子驱动。 CCND2过表达的hipsc- 衍生的心肌细胞（HIPSC-CCND2OECM）表现出增加的细胞周期活性和细胞增殖 与表达野生型CCND2水平（HIPSC-CCND2WTCMS）的普通幼稚的HIPSC-CM相比。在 心肌梗死的小鼠模型（MI），刻有三倍的雕刻细胞数量 与接受HIPSC-CCND2WTCM的HIPSC-CCND2OECM相比，MI和移植后4周 导致梗死大小和改善心脏功能的明显较小。这些数据表明转基因 HIPSC-CM图形中的CCND2过表达构成了一种可行的方法来增强植入和恢复 缺血性心脏病的功能。 该提案将使用HIPSC-CCND2OECM开发一种新型的人类心肌斑（被指定为HCMP- CCND2OECMS），评估其能力在长期内持续地震惊的心脏不断地震惊的心脏 最终取代疤痕组织，测试其安全性并在大型动物模型中转化潜力（该模型 猪MI模型）。在移植的HCMP中，改善了慢性梗塞的猪心的心脏功能， 该提案还确定了这种功能改进是否归因于Rebuscularization或其他 机制。特别是，我们旨在解决尚未解决但非常重要的问题，质疑HCMPS是否存在 通过猪MI模型中的直接供体主持耦合改善心脏功能。我们的长期目标是发展心 可以转化为人类的再生策略。提出了两个具体目标。 具体目标1将检验以下假设：这种新型的HCMP不断地雷神化的心肌， 替换透经疤痕组织，并改善梗塞猪心脏的心脏功能。我们将确定（i） 斑块移植对心脏结构和功能的影响，以及（ii）斑块移植的安全性 （抑制诱导性心律不齐和形成肿瘤的风险）。 特定目标2检验了功能变化的大小与数量正相关的假设 供体细胞来源的CM。我们将确定（i）供体细胞存活是否需要持续改善需要 心脏功能以及（ii）至少部分地介导心脏功能的改善是否是由 移植供体肌细胞的机电耦合。