Reprogramming using small molecules

描述（由申请人提供）：四种转录因子（Oct 4、Sox 2、cMyc和Klf 4）的组合活性可以将成体细胞重编程为诱导多能干（iPS）细胞。这些重新编程的细胞应该为移植医学、药物发现和人类疾病研究提供无限的遗传定制细胞类型。不幸的是，用于递送重编程因子的方法已经引起了关于所得干细胞的未来效用的担忧，或者可能与临床相容的干细胞系的工业规模生产不相容。在这里，我们建议确定小分子的作用机制，我们已经证明，可以通过增加其效率或允许省略一个或多个重编程因子来促进重编程过程。我们的最终目标是确定一种化学制剂，可以单独将成体细胞重新编程为多能状态。这些重新编程的细胞将不受遗传操纵，并将成为生物医学应用的最佳多能细胞。 具体目标是：目的1）确定新鉴定的小重编程分子在重编程过程中取代Klf 4的机制;目的2）确定新鉴定的可取代Oct 4的小重编程分子在重编程过程中的作用机制;目的3）为了确定我们已经鉴定的重编程分子是否可以协同作用以替代多种重编程转录因子，从而使我们更接近于一种完全化学的方法来重新编程。

DESCRIPTION (provided by applicant): The combined activity of four transcription factors (Oct4, Sox2, cMyc and Klf4) can reprogram adult cells into induced pluripotent stem (iPS) cells. These reprogrammed cells should provide a limitless supply of genetically tailored cell types for transplantation medicine, drug discovery and the study of human disease. Unfortunately, the methods used to deliver reprogramming factors have either raised concerns regarding the future utility of the resulting stem cells, or may not be compatible with industrial scale production of clinically compatible stem cell lines. Here, we propose to determine the mechanisms of action of small molecules that we have shown can facilitate the reprogramming process by either increasing its efficiency or by allowing the omission of one or more reprogramming factors. Our final goal is to move towards the identification of a chemical formulation that can alone reprogram adult cells to a pluripotent state. These reprogrammed cells would be free of genetic manipulation and would be the optimal pluripotent cells for biomedical applications. Specifically the aims are: aim 1) To determine the mechanisms by which newly identified small reprogramming molecules act to replace Klf4 in the reprogramming process; aim 2) To determine the mechanisms by which newly identified small reprogramming molecules that can replace Oct4 act in the reprogramming process; aim 3) To determine whether the reprogramming molecules that we have identified can act synergistically to replace multiple reprogramming transcription factors, thus moving us closer to a completely chemical method for reprogramming.