Human Chemically induced Pluripotent Stem Cells (CiPSC) generated Cardiomyocytes

描述(申请人提供)：通过诱导成人体细胞直接重编程产生的人类干细胞，称为诱导多能干细胞(IPS)细胞，为研究人类疾病特定/个性化模型提供了“范式转换”机会，并为实际的自体细胞疗法、再生医学和预测毒理学应用提供了个性化的可再生细胞来源。本提案中概述的研究旨在探索CiPSC在生成个性化心肌细胞方面的应用，这些细胞可用于心血管疾病的建模、毒理学测试以及最终用于心血管再生细胞治疗。我们建议使用一种新的稳健而高效的iPS方法，该方法使用一种小分子诱导剂的鸡尾酒，称为化学诱导多能干细胞(CiPSC)，消除了对任何外源基因转导的需要(即，其他人所描述的iPS方法的主要障碍)。在这项建议中，这种方法的可行性将通过使用成熟的从iPS和hESC体外获得心肌细胞的程序获得心肌细胞(CiPSC-CM)来证明。这些CiPSC是在不对其他DNA载体诱导的iPS方法所观察到的细胞和分子机制进行任何异常和永久性修改的情况下获得的。我们的长期目标(第二阶段及以后)是利用CiPSC-CMS产生宿主相容的CM替代疗法，产生体外疾病特异性模型，并促进在动物和人类模型系统中进行体外治疗和毒性筛选。 公共卫生相关性：通过诱导成人体细胞直接重编程产生的人类干细胞，称为诱导多能干细胞(IPS)，为研究人类疾病特定/个性化模型提供了“范式转换”机会，并为实际的自体细胞疗法、再生医学和预测毒理学应用提供了个性化的可再生细胞来源。然而，使用iPS细胞作为开发此类患者和疾病特异性模型系统的工具的可能性充其量仍具有挑战性，由于该领域的缺陷，仍存在明显的未得到满足的需求。本提案中概述的研究旨在探索化学诱导多能干细胞(CiPSC)在生成个性化心肌细胞方面的应用，这些细胞可用于心血管疾病的建模、毒理学测试以及最终用于心血管再生细胞治疗。CiPSC消除了任何外源基因转导的需要，这是迄今为止其他人描述的iPS方法的主要障碍。

DESCRIPTION (provided by applicant): Human stem cells generated by induced direct reprogramming of adult somatic cells, termed induced pluripotent stem (iPS) cells, offer "paradigm shifting" opportunities for studying human disease specific/ personalized models, and provide a personalized renewable source of cells for practical autologous cell therapies, regenerative medicine, and predictive toxicology applications. Studies outlined in this proposal are designed to explore application of CiPSC for the generation of personalized cardiomyocytes which can be used for modeling of cardiovascular diseases, toxicology testing and ultimately for cardiovascular regenerative cell therapy. We propose to use a novel robust and efficient iPS methodology which employs a cocktail of small molecule inducers, termed "Chemically induced Pluripotent Stem (CiPSC), eliminating the need for any exogenous gene transduction (i.e. a major impediment of iPS methodologies described by others). In this proposal, feasibility of this approach will be demonstrated via derivation of cardiomyocytes (CiPSC-CM) using well established procedures for in-vitro derivation of cardiomyocytes from iPS and hESC. These CiPSC are obtained without any abnormal and permanent modifications to the cellular and molecular machinery typically observed by other DNA vector induced iPS methodologies. Our long-term goals (Phase II and beyond) are to use CiPSC-CMs in generating host- compatible CM replacement therapy, producing in-vitro disease-specific model and facilitating in-vitro therapeutic and toxicity screen in animal and human model systems. PUBLIC HEALTH RELEVANCE: Human stem cells generated by induced direct reprogramming of adult somatic cells, termed induced pluripotent stem (iPS) cells, offer "paradigm shifting" opportunities for studying human disease specific/ personalized models, and provide a personalized renewable source of cells for practical autologous cell therapies, regenerative medicine, and predictive toxicology applications. The possibility of using iPS cells as a tool for development of such patient and disease specific model systems, however, remains at best challenging and stills a clear unmet need due to the shortcomings in this field. Studies outlined in this proposal are designed to explore application of Chemically induced Pluripotent Stem Cells (CiPSC) for the generation of personalized cardiomyocytes which can be used for modeling of cardiovascular diseases, toxicology testing and ultimately for cardiovascular regenerative cell therapy. CiPSC eliminates the need for any exogenous gene transduction, which is a major impediment of iPS methodologies described to date by others.