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：逆向工程人类神经系统疾病）。病毒或质粒引入转录因子四重奏被证明是一种有效的策略，可以触发体细胞重编程为诱导多能干细胞（iPS），而不需要胚胎或卵子。然而，使用转基因的基因操作代表了将这些iPS细胞用于治疗应用的一个严重障碍。虽然转基因基因的再激活可能导致肿瘤发生，但转基因基因的泄漏表达可能抑制iPS细胞分化，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确定不需要基因转移就能诱导内源性多能调节因子的小分子。此外，利用现有技术通过病毒转导或质粒转染重编程因子来生成iPS细胞是一个效率非常低的过程。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，并鉴定提高重编程效率的化合物。本研究将为多能性人类干细胞系的高效衍生提供新方法，并为多能性干细胞的产生开辟一条新的途径。这些化学衍生的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。

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.