Generation of Induced Pluripotent Stem Cells with Novel Small Molecule Regulator

描述（由申请人提供）：该申请涉及广泛的挑战领域（14）：干细胞，并满足一些特定的挑战主题：14- eb -101：生成多能干细胞的合成递送系统14- pk -101：诱导多能干细胞-疾病的细胞和人源化小鼠模型14- pk -102：干细胞或祖细胞编程方法的发现06- gm102：一系列研究报道，通过引入一些转录因子，可以将完全分化的体细胞重编程为未分化的多能干细胞。这些诱导多能干细胞（iPS）与胚胎干细胞（ES）具有几乎相同的多能性。然而，对于任何将外源遗传因素引入体细胞的方法，特别是涉及到可能诱发肿瘤或其他不良反应的病毒整合或外源基因时，都存在严重的担忧。虽然最近已经开发出非病毒方法来产生人类iPS细胞，但细胞的来源仅限于人类胚胎成纤维细胞，因此不能用于产生用于研究的疾病特异性iPS细胞或用于细胞治疗的自体iPS细胞。由于这些是iPS细胞相对于胚胎干细胞的主要优势，因此需要额外的来源和更安全的重编程方法。在撰写本文时，我们已经成功地使用小分子将小鼠成纤维细胞重新编程为iPS细胞。此外，我们还利用病毒方法从正常志愿者和冠心病（CAD）患者的外周血中生成了iPS细胞。因此，在本提案中，我们将寻求开发新的方法来产生人类iPS细胞，使用外周血作为来源，结合小分子表观遗传调节剂。我们预计这项研究将对人类iPS细胞的生成方法产生根本性的改变，并对iPS细胞的疾病研究和临床应用产生巨大的影响。

DESCRIPTION (provided by applicant): This application addresses broad Challenge Area (14): Stem Cells, and meets a few specific Challenge Topics: 14-EB-101: Synthetic Delivery Systems for Generating Pluripotent Stem Cells 14-DK-101: Induced pluripotent stem cells - cellular and humanized mouse models of disease 14-DK-102: Discovery of methods to program stem or progenitor cells 06-GM-102: Chemist/biologist collaborations facilitating tool development A series of studies reported that reprogramming of fully differentiated somatic cells into undifferentiated, pluripotent stem cells could be achieved by introducing a number of transcription factors. These induced pluripotent stem (iPS) cells have almost identical pluripotency to embryonic stem (ES) cells. However, there are serious concerns about any approach in which exogenous genetic factors are introduced into somatic cells, and especially when viral integration or foreign genes, which may induce tumors or other adverse effects, are involved. Although non-viral approaches have recently been developed to generate human iPS cells, the source of cells was limited to human embryonic fibroblasts and therefore could not be used to generate disease-specific iPS cells for research or autologous iPS cells for cell therapy. As these are the main advantages of iPS cells over ES cells, additional sources and safer reprogramming methods are needed. At the time of writing, we have successfully reprogrammed mouse fibroblasts into iPS cells using only small molecules. In addition, we have generated iPS cells from peripheral blood of normal volunteers and coronary artery disease (CAD) patients using viral approaches. Accordingly, in this proposal, we will seek to develop novel methods to generate human iPS cells using peripheral blood as a source in combination with small molecular epigenetic regulators. We anticipate that this study will effect a fundamental change in the method of generating human iPS cells and will have enormous impact on disease investigation and clinical application of iPS cells. PUBLIC HEALTH RELEVANCE: A series of studies reported that reprogramming of fully differentiated somatic cells into undifferentiated, pluripotent stem cells, referred to as induced pluripotent stem (iPS) cells, could be achieved by forced expression of pluripotency-related transcription factors. However, the current technology is limited to the use of genetic material (viral or plasmid vectors) for delivery and fibroblasts for the source of parental cells. Accordingly, in this proposal, we will seek to develop novel methods to generate human iPS cells using combinations of small molecular epigenetic regulators with peripheral blood as the cell source.