Project 1: Establishment & maintenance of the pluripotent state

发育生物学和生物医学界长期以来一直有兴趣了解 具有相同基因组的细胞如何建立和维持显著不同的行为和基因表达 模式. 试图定义足以将一种细胞类型重编程为另一种细胞类型的特定因子， 鉴定的单个或有限的转录因子组合。例如，胚胎干细胞的引入- 表达的转录因子Oct 4/Sox 2/Klf 4/c-Myc足以将成纤维细胞重编程为多能性细胞。 状态转录因子的异位表达可能是一种通用和实用的方法， 在任意细胞状态之间重新编程。但是，我们从初步研究中也知道， 这些因素本身无法引起完全的转换，我们发现了大量的异质性， 一旦多能状态建立，就可以改变。 在这里，我们的目标是大大提高我们对重编程和多能性本身的一般理解。 为了实现这一点，我们将首先利用重编程过程的方向性， 确定细胞环境相关的重塑能力（aim 1），然后生成多维数据 描述了多能群体（目标2），并最终建立了工具，使我们能够进一步表征 存在于多能细胞培养物中的亚群（aim 3）。拟议的研究提供了一个机会， 显著改变和加速细胞重编程在基础、转化和临床上的应用， 应用. 相关性（参见说明）： 拟议的研究工作的目的是提供一个详细的机制了解的关键蛋白质因素 影响细胞状态转变（体细胞到多能性）以及随后的维持， 多能国家更好地理解多能性的建立和维持， 改变生物医学研究和医疗服务。

The developmental biology and biomedical communities have had a long-standing interest in understanding how cells with identical genomes establish and maintain remarkably different behaviors and gene expression patterns. Attempts to define specific factors that are sufficient for reprogramming one cell type into another have identified single, or limited combinations of, transcription factors. For example, introduction of the ES cell- expressed transcription factors Oct4/Sox2/Klf4/c-Myc are sufficient to reprogram fibroblasts to a pluripotent state. Ectopic expression of transcription factors is likely to be a general and practical method for reprogramming between arbitrary cellular states. However, we also know from our preliminary studies that the factors themselves are unable to induce the full conversion and we find a great deal of heterogeneity and variability once the pluripotent state is established. Here we aim to dramatically advance our general understanding of reprogramming and pluripotency itself. To accomplish this we will first take advantage of the directionality of the reprogramming process and determine the cell context dependent remodeling capacity (aim 1), and then generate multidimensional data that describe the pluripotent population (aim 2), and finally built tools that enable us to further characterize subpopulations present within pluripotent cell cultures (aim 3). The proposed study presents an opportunity to dramatically alter and accelerate the utility of cellular reprogramming for basic, translational and clinical applications. RELEVANCE (See instructions): The proposed research effort aims to provide a detailed mechanistic understanding of the key protein factors that influence the cell state transitions (somatic to pluripotent) as well as the subsequent maintenance of the pluripotent state. A better understanding of the establishment and maintenance of pluripotency could transform biomedical research and health care delivery.