Generation of human iPS cells by chemically defined conditions

通过化学定义的条件生成人类 iPS 细胞

• 批准号: 8145688
• 负责人: SHENG DING
• 金额: $ 37.39万
• 依托单位: J. DAVID GLADSTONE INSTITUTES
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-09-17 至 2015-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8145688
• 关键词: Autologous; Biochemical; Biological Assay; Cardiovascular Diseases; Cells; Chemicals; Development; Diabetes Mellitus; Disease; Ectopic Expression; Epigenetic Process; Exposure to; Generations; Genetic Materials; Human; In Vitro; Kinetics; Lead; Malignant Neoplasms; Methods; Molecular; Neurodegenerative Disorders; Peptides; Principal Investigator; Process; Production; Proteins; Recombinant Proteins; Recombinants; Regulation; Reporting; Rodent; Series; Signal Transduction; Somatic Cell; Source; Specificity; Stem cells; Structure-Activity Relationship; Testing; Therapeutic; Tissues; Viral; base; c-Myc Staining Method; cell type; genetic manipulation; high throughput screening; human disease; improved; in vivo; induced pluripotent stem cell; novel; organ regeneration; pluripotency; programs; public health relevance; self-renewal; small molecule; stem cell fate; tool; transcription factor

DESCRIPTION (provided by applicant): Although stem cells hold considerable promises for the treatment of a number of devastating diseases (e.g. cardiovascular diseases, neurodegenerative diseases, diabetes and cancers), obstacles such as control of stem cell fate/function, immuno-rejection, and limited cell sources must be overcome before their therapeutic potentials can be realized. Recent studies have suggested that tissue-specific somatic cells may overcome their intrinsic lineage-restriction to de-differentiate or trans-differentiate (i.e. reprogramming) upon exposure to a specific set of signals (or ectopic expression of master transcription factors) in vitro and in vivo. Reprogramming of various rodent and human somatic cells to pluripotent ESC-like cells (i.e. induced pluripotent/iPS cells) has recently been achieved by viral transduction of four transcription factors (e.g. Oct4, cMyc, Sox2 and Klf4). More recently, we have reported that small molecules can replace certain transcription factors as well as substantially improve reprogramming efficiency and kinetics in generating iPS cells, and developed a method of generating and using recombinant cell-penetrating proteins to generate iPS cells without using any genetic materials and genetic manipulation (please see our preliminary results). With those proof-of-principle demonstrations, we propose to further develop/optimize the chemically defined, protein and small molecule-based reprogramming process in human cells to have highly robust and efficient method and to further in-depth characterize those cells. PUBLIC HEALTH RELEVANCE: Reprogramming of various rodent and human somatic cells to pluripotent ESC-like cells (i.e. induced pluripotent/iPS cells) has recently been achieved by viral transduction of four transcription factors (e.g. Oct4, cMyc, Sox2 and Klf4). More recently, we have reported that small molecules can replace certain transcription factors as well as substantially improve reprogramming efficiency and kinetics in generating iPS cells, and developed a method of generating and using recombinant cell-penetrating proteins to generate iPS cells without using any genetic materials and genetic manipulation (please see our preliminary results). With those proof-of-principle demonstrations, we propose to further develop/optimize the chemically defined, protein and small molecule-based reprogramming process in human cells to have highly robust and efficient method and to further in-depth characterize those cells. Collectively, the studies described in this proposal will provide novel chemical tools for producing unlimited amount of (autologous) pluripotent cells from differentiated/lineage-restricted cells for various applications as well as studying the underlying molecular mechanisms of pluripotency and epigenetic regulations, and may ultimately facilitate development of small molecule therapeutics to treat human diseases and stimulate tissue/organ regeneration in vivo.

描述（由申请人提供）：尽管干细胞对多种毁灭性疾病的治疗（例如心血管疾病，神经退行性疾病，糖尿病和癌症）的治疗有很大的承诺，但是诸如干细胞命运/功能的控制，限制细胞的障碍必须在其更具范围的潜在范围之前实现。最近的研究表明，组织特异性的体细胞可以在暴露于特定的信号（或主转录因子的异位表达）时，可以克服其内在的谱系限制，以去分化或反差异（即重编程）。 最近通过病毒转导四个转录因子（例如Oct4，Cmyc，Sox2和Klf4）来实现各种啮齿动物和人体细胞对多能ESC样细胞（即诱导多能/IPS细胞）的重编程。最近，我们报告说，小分子可以替代某些转录因子，并在产生IPS细胞时实质上提高了重编程效率和动力学，并开发了一种生成和使用重组细胞培训蛋白来生成IPS细胞的方法，以生成IPS细胞，而无需使用任何遗传材料和遗传操纵（请参阅我们的初步结果）。通过这些原则证明，我们建议在人类细胞中进一步开发/优化基于化学定义的，蛋白质和基于小分子的重编程过程，以具有高度鲁棒和有效的方法，并进一步深入地表征这些细胞。 公共卫生相关性：最近通过病毒转导四个转录因子（例如OCT4，CMYC，SOX2和KLF4）来实现各种啮齿动物和人类体细胞对多能ESC样细胞（即诱导多能/IPS细胞）的重编程。最近，我们报告说，小分子可以替代某些转录因子，并在产生IPS细胞时实质上提高了重编程效率和动力学，并开发了一种生成和使用重组细胞培训蛋白来生成IPS细胞的方法，以生成IPS细胞，而无需使用任何遗传材料和遗传操纵（请参阅我们的初步结果）。通过这些原则证明，我们建议在人类细胞中进一步开发/优化基于化学定义的，蛋白质和基于小分子的重编程过程，以具有高度鲁棒和有效的方法，并进一步深入地表征这些细胞。 总的来说，该提案中描述的研究将提供新的化学工具，用于从分化/谱系限制的细胞中生产无数量的（自体）多能细胞，以用于各种应用，并研究多能性和表观远期法规的潜在分子机制，并可能最终促进小分子的构造和刺激人类疾病的发展。