Chemically Induced Pluripotent Stem (CiPS) Cells Alzheimer's Patient Specific Cel

描述(由申请人提供)：由于缺乏真正的体外模型，研究阿尔茨海默病(AD)等人类疾病的病理生理学的努力受到阻碍。通过诱导成体细胞直接重编程产生的干细胞，被称为诱导多能干细胞(IPS)，通过为研究人类疾病提供特定/个性化的模型，以及为实际的自体细胞治疗和再生医学应用提供个性化的可再生细胞来源，从而避免免疫排斥，从而提供了“范式转换”的机会。然而，使用iPS细胞作为开发此类AD患者特定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，仍是一个明显的未得到满足的需求。这项提案中概述的研究旨在探索这一未得到满足的需求的解决方案，最终目标是开发AD患者特有的细胞，作为研究这种疾病的病理生理学和药物测试的工具。我们建议使用一种新的稳健和高效的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(CiPSTM)细胞，消除了对任何外源基因转导的需要(即，其他人描述的iPS方法的主要障碍)。在这项第一阶段研究中，将通过对少量AD患者的皮肤活检、成纤维细胞样本进行CIPS衍生来证明该方法的可行性。这些CIPS患者特异性细胞将被获得，而不会对其他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不正常和永久性的修改。拟议研究开发的工具和技术可直接应用于研究其他神经退行性疾病和年龄相关疾病的分子和细胞途径。 公共卫生相关性：由于缺乏真正的体外模型，研究阿尔茨海默病等人类疾病的病理生理学的努力受到阻碍。干细胞的医学和生物技术潜力已被认为是了解基本疾病机制、筛选药物发现以及组织工程治疗退行性疾病和致残性损伤的前所未有的机会。特别是，通过诱导成体细胞直接重编程产生的干细胞，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”的机会，并为实际的自体细胞治疗和再生医学应用提供了个性化的可再生细胞来源，从而避免了免疫排斥。我们建议使用一种新的可靠而有效的诱导多能干细胞的方法，该方法使用小分子诱导剂的鸡尾酒，被称为“化学诱导多能干细胞(CiPSTM)细胞，它不需要任何外源基因转导。患者特异性的多能干细胞被获得，而不需要对细胞和分子机制进行任何异常和永久性的修改(这是迄今为止报道的所有其他DNA载体介导的诱导多能干细胞的主要缺陷)。该研究开发的工具和技术直接应用于其他神经退行性疾病和年龄相关疾病的分子和细胞通路的研究。

DESCRIPTION (provided by applicant): Efforts to investigate the pathophysiology of human diseases such as Alzheimer's disease (AD) are hampered by the lack of genuine in-vitro models. Stem cells generated by induced direct reprogramming of adult somatic cells, termed induced pluripotent stem (iPS) cells, offer "paradigm shifting" opportunities by providing specific/ personalized models for studying human disease, and personalized renewable source of cells for practical autologous cell therapies and regenerative medicine applications, that avoid immune rejection. The possibility of using iPS cells as a tool for development of such AD patient specific model systems, however, remains at best challenging and still a clear unmet need due to the shortcomings in this field. Studies outlined in this proposal are designed to explore resolution of this unmet need with the ultimate goal of developing AD patient specific cells as a tool for study of the pathophysiology of this disease and drug testing. We propose to use a novel robust and efficient iPS methodology which employs a cocktail of small molecule inducers, termed "Chemically induced Pluripotent Stem (CiPSTM) cells, eliminating the need for any exogenous gene transduction (i.e. a major impediment of iPS methodologies described by others). In this Phase I study, feasibility of this approach will be demonstrated via CiPS derivation of a small number of AD patients', skin biopsy, fibroblasts samples. These CiPS patient specific cells will be obtained without any abnormal and permanent modifications to the cellular and molecular machinery typically observed by other DNA vector induced iPS methodologies. Tools and technologies developed by the proposed study have direct applications to the study of molecular and cellular pathways of other neurodegenerative and age related diseases. PUBLIC HEALTH RELEVANCE: Efforts to investigate the pathophysiology of human diseases such as Alzheimer's disease are hampered by the lack of genuine in-vitro models. Medical and biotechnological potential of stem cells have been recognized as unprecedented opportunities for understanding basic disease mechanisms, screens for drug discovery, and tissue engineering for degenerative diseases and crippling injuries. In particular, 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 and regenerative medicine applications, that avoid immune rejection. We propose to use a novel robust and efficient iPS methodology that uses a cocktail of small molecule inducers, termed "Chemically induced Pluripotent Stem (CiPSTM) cells, which eliminates the need for any exogenous gene transduction. Patient specific pluripotent cells are derived, without any abnormal and permanent modifications to the cellular and molecular machinery (i.e. a major shortcoming of all other DNA vector mediated iPS inductions reported to date). Tools and technologies developed by the proposed study have direct applications to the study of molecular and cellular pathways of other neurodegenerative and age related diseases.