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对理解 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新编程为另一种细胞类型的特定因素 识别出单一的或有限的转录因子组合。例如，ES细胞的引入- 表达的转录因子Oct4/Sox2/Klf4/c-Myc足以将成纤维细胞重新编程为多能细胞 州政府。转录因子的异位表达可能是一种通用和实用的方法 在任意蜂窝状态之间重新编程。但是我们也从我们的初步研究中得知， 这些因素本身不能诱导完全转化，我们发现存在很大的异质性和 一旦建立了多能性状态，就会发生变化。 在这里，我们的目标是极大地提高我们对重新编程和多能性本身的总体理解。 为了实现这一点，我们将首先利用重新编程过程的方向性，并 确定依赖于单元格上下文的重塑能力(目标1)，然后生成多维数据 描述了多能人口(目标2)，并最终建立了工具，使我们能够进一步表征 多能细胞培养中存在亚群(目标3)。拟议的研究提供了一个机会 极大地改变和加速基础、翻译和临床细胞重编程的效用 申请。 相关性(请参阅说明)： 拟议的研究工作旨在提供对关键蛋白质因素的详细机制理解。 这影响了细胞状态的转变(从体细胞到多能性)以及随后细胞状态的维持 多能状态。更好地理解多能性的建立和维持可以 转变生物医学研究和医疗保健服务。

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.