Induced Pluripotent Stem Cells in the Understanding and Treatment of Heart Diseas

描述(由申请人提供)： 在美国，每年有100多万美国人患有急性心肌梗死，在幸存者中，有500万人患有心力衰竭。此外，细胞谱系确定或形态发生的缺陷是先天性心脏畸形的基础，先天性心脏畸形是人类最常见的出生缺陷。在美国，先天性心脏病的幸存者超过100万人，他们也经常患有心力衰竭。不幸的是，心脏在受伤后几乎没有再生能力。最近人类诱导多能干细胞(IPS)的发现为人类疾病的新方法打开了大门，包括开发用于疾病机制和药物发现的人类细胞模型，以及基于自体细胞的治疗的可能性。我们建议在格拉德斯通研究所和斯坦福大学组建一个研究团队，以开发和利用IPS细胞在心脏病治疗和理解中的潜力。避免DNA基因组整合的IPS生成方法正在迅速发展，但在人类使用iPS细胞之前仍需要改进；这一障碍将在本应用中解决。随着IPS细胞生成方法的改进，该团队将共同努力，更有效地生成iPS来源的心肌细胞，用于未来的治疗，利用他们在染色质重塑和微RNA(MiRNA)生物学以及G蛋白偶联受体信号方面的专业知识。该团队将使用细菌人工染色体(BAC)策略，为心脏分化的进展阶段产生带有荧光标记的iPS细胞系。我们还将尝试将体细胞直接重新编程为心脏祖细胞。通过我们在斯坦福大学的合作伙伴，我们将在啮齿动物和大型动物身上检查体内细胞的存活和植入。将产生疾病特异的iPS细胞，以揭示人类祖细胞生物学的新方面。这个多学科团队将为NHLBI前体细胞联盟带来广泛和关键的专业知识，努力积极利用iPS细胞在心脏病方面的前景和潜力。与我们中心内的斯坦福大学团队的互动将协同并利用每个研究小组在与iPS细胞相关的重点工作上的特定优势。其具体目标是：1)开发无整合高效的人IPS细胞的生成方法，用于未来的细胞治疗；2)开发高效的人IPS细胞的定向分化和直接重编程的方法；3)开发用于未来细胞治疗的人IPS细胞。开发在心血管疾病动物模型中使用IPS细胞来源的心脏祖细胞的方法。利用疾病特异性IPS细胞发现人类心脏祖细胞生物学和心血管疾病机制。

DESCRIPTION (provided by applicant): Over 1 million Americans suffer acute myocardial infarctions each year in the US, and among the survivors, 5 million are afflicted with heart failure. In addition, defects in cell lineage determination or morphogenesis underlie congenital heart malformations, the most common human birth defect. Survivors of congenital heart disease, who number over 1 million in the US, also often suffer from heart failure. Unfortunately, the heart has little regenerative capacity after injury. The recent discovery of human induced pluripotent stem (IPS) cells has opened the door for novel approaches to human disease, including the development of human cellular models for disease mechanisms and drug discovery, along with the potential for autologous cell-based therapies. We propose to assemble a team of investigators at the Gladstone Institutes and Stanford University to develop and capitalize on the potential of IPS cells in the treatment and understanding of heart disease. Methods of IPS generation avoiding genomic integration of DNA are developing rapidly, but continue to require refinement before use of iPS cells in humans; this hurdle will be addressed in this application. As methods for generating IPS cells are improved the team will work together to more efficiently generate iPS-derived cardiac cells for future therapy, capitalizing on their expertise in chromatin remodeling and microRNA (miRNA) biology and G-protein coupled receptor signaling. The team will generate iPS cell lines with fluorescent markers for progressive stages of cardiac differentiation using bacterial artificial chromosome (BAC) strategies. We will also attempt to reprogram somatic cells directly into cardiac progenitors. Survival and engraftment of cells in vivo will be examined in rodents and in large animals through our partners at Stanford. Disease-specific iPS cells will be generated to reveal novel aspects of human progenitor cell biology. This multidisciplinary team will bring broad and critical expertise to the NHLBI Progenitor Cell Consortium in an effort to aggressively capitalize on the promise and potential of iPS cells for heart disease The interaction with the Stanford group within our Hub will synergize and leverage the specific strengths of each group of investigators on the focused effort related to iPS cells. The specific aims are: 1) To develop integration-free and efficient methods of human IPS cell generation for future cell-based therapies; 2) To develop efficient directed differentiation of human IPS cells and methods of direct reprogramming; 3). To develop methods to use IPS cell-derived cardiac progenitors in animal models of cardiovascular disease and 4). To use disease-specific IPS cells for discovery of human cardiac progenitor biology and cardiovascular disease mechanisms.