Derivation of human pluripotent stem cells using small molecules

描述(由申请人提供)：本申请涉及广泛的挑战领域(14：干细胞)和特定的挑战主题(14-NS-101：逆向工程人类神经疾病)。病毒或质粒导入转录因子四元组被证明是一种有效的策略，可以触发体细胞重新编程为诱导的多能干细胞(IPS)，而不需要胚胎或卵子。然而，使用转基因进行基因操作是将这些iPS细胞用于治疗应用的一个严重障碍。虽然转基因基因的重新激活可能导致肿瘤的发生，但转基因的泄漏表达可能会抑制iPS细胞的分化，增加未成熟畸胎瘤形成的风险。解决这一问题的一种方法是在不进行基因转移的情况下识别能够诱导内源性多能调节的小分子。此外，利用现有技术通过病毒转导或重编程因子的质粒转染来产生iPS细胞是一个效率很低的过程。这项研究的主要目标是利用小分子获得无转基因的人类iPS细胞，并鉴定能够提高重编程效率的化合物。这项研究将为高效获得多能人类干细胞系开发新的方法，并为产生患者和疾病特异性多能干细胞开辟一条新的途径。这些化学来源的iPS细胞将成为发育生物学、药物发现和再生医学的宝贵工具。 公共卫生相关性：以无病毒和无转基因的方式化学衍生人类诱导性多能干细胞将开辟一条高效产生患者和疾病特异性多能干细胞的新途径。这些未经基因改造的iPS细胞将应用于基于干细胞的替代疗法，用于治疗神经退行性疾病，如阿尔茨海默氏症和帕金森氏症，以及脑损伤。

DESCRIPTION (provided by applicant): This application addresses broad Challenge Area (14: Stem Cells) and specific Challenge Topic (14-NS-101: Reverse Engineering Human Neurological Disease). Viral or plasmid introduction of a transcription factor quartet is proven a powerful strategy to trigger reprogramming somatic cells into induced pluripotent stem (iPS) cells without the need of embryos or eggs. However, genetic manipulation using transgenes represents a serious hurdle to the use of these iPS cells for therapeutic application. While reactivation of trangenes could lead to tumorigenesis, leaky expression of transgenes may inhibit iPS cell differentiation, increasing the risk of immature teratoma formation. One way to solve this problem is to identify small molecules that induce endogenous pluripotent regulators without gene transfer. Furthermore, generation of iPS cells using existing technology by viral transduction or plasmid transfection of the reprogramming factors is a process with very low efficiency. The main goal of this research is to derive transgene-free human iPS cells using small molecules and to identify compounds that enhance reprogramming efficiency. This study will lead to the development of new methods for the derivation ofpluripotent human stem cell lines with high efficiency and open a new avenue to generate patient- and disease-specific pluripotent stem cells. These chemically derived iPS cells will become valuable tools for developmental biology, drug discovery, and regenerative medicine. PUBLIC HEALTH RELEVANCE: Chemical derivation of human iPS cells in virus-free and transgene-free means will open a new avenue to generate patient- and disease-specific pluripotent stem cells with high efficiency. These genetically unmodified iPS cells will be applicable in stem cell-based cell replacement therapies for the treatment of neurodegenerative diseases, such as Alzheimer's and Parkinson's diseases, and brain injuries.