Human Pluripotent Stem Cell and Progenitor Models of Cardiac and Blood Diseases

描述（由申请人提供）：晚期心力衰竭是一种主要的未满足的临床需求，由存活或功能完整的心肌细胞丧失引起。尽管有最佳的药物治疗，心力衰竭是美国死亡的主要原因。从可再生的患者特异性来源产生功能性心肌组织将允许开发用于途径鉴定和药物开发的疾病特异性细胞模型。这也将为治疗性心脏再生医学奠定基础。我们建议整合心脏干细胞生物学和开发（Chien/Wu），人类ES和IPS技术（Melton）和心脏组织工程（帕克）四个领先实验室的独特专业知识，试剂和方案，以产生患者特异性心肌组织作为人类心血管疾病的细胞模型，并作为基于细胞的再生治疗的基础。因此，我们拟议项目的具体目标如下。目的1：确定控制心血管祖细胞（CVP）扩增和分化的非细胞自主信号通路。我们推测，特定的非细胞自主信号通路控制小鼠CVP的扩增和分化。目的2：确定小鼠iPS衍生的CVP是否重现正常心脏发育，并可用于体外产生功能性心肌组织。我们假设IPS衍生的心脏祖细胞在发育潜力和功能特征方面与ESC衍生的心脏祖细胞相似。目的3：从人ES和IPS细胞中分离和鉴定CVP。我们假设在人ES和iPS细胞分化过程中，心肌细胞是由心脏祖细胞定向分化为CVP产生的，CVP然后分化为完全成熟的心室肌细胞。目标4：生成人类心血管疾病的细胞模型，如X连锁心肌病（即由肌营养不良蛋白突变引起的心肌病，如Duchenne和Becker肌营养不良症）。我们假设疾病特异性CVP将重现病变心肌表型以及病变心肌表型。这将作为遗传途径发现，药物设计和再生心血管医学的平台。

DESCRIPTION (provided by applicant): Advanced heart failure represents a major unmet clinical need, arising from the loss of viable or fully functional cardiac muscle cells. Despite optimum drug therapy, heart failure is a leading cause of mortality in America. Generating functional myocardial tissue from a renewable patient-specific source would allow for the development of disease specific cellular models for pathway identification and drug development. It would also lay the foundation for therapeutic cardiac regenerative medicine. We propose to integrate the unique expertise, reagents, and protocols from four leading laboratories in cardiac stem cell biology and development (Chien/Wu), human ES and IPS technology (Melton), and cardiac tissue engineering (Parker), to generate patient-specific myocardial tissue as cellular models for human cardiovascular disease, and as a foundation for cell-based regenerative therapy. Accordingly, the specific aims of our proposed project are the following. AIM 1: Identify non-cell autonomous signaling pathways that control cardiovascular progenitor cell (CVP) expansion and differentiation. We hypothesize that specific non-cell autonomous signaling pathways control murine CVP expansion and differentiation. AIM 2: Determine if murine iPS-derived CVPs recapitulate normal cardiac development and can be used to generate functional myocardial tissue in vitro. We hypothesize that IPS derived cardiac progenitors are similar to ESC-derived cardiac progenitors in developmental potential and functional characteristics. AIM 3: Isolate and characterize CVPs from human ES and IPS cells. We hypothesize that during human ES and iPS cell differentiation, cardiac myocytes are generated by the commitment of cardiac progenitors into CVPs which then differentiate into fully mature ventricular myocytes. AIM 4: Generate cellular models for human cardiovascular disease such as X-linked cardiomyopathy (i.e. cardiomyopathies caused by dystrophin mutations such as those in Duchenne's and Becker's muscular dystrophy). We hypothesize that disease specific CVP will recapitulate diseased myocardial phenotype, as well as diseased myocardial phenotype. This will serve as a platform for genetic pathway discovery, drug design, and regenerative cardiovascular medicine.