Project 1: Establishment & maintenance of the pluripotent state

项目一：建立

• 批准号: 8535279
• 负责人: Alexander Meissner
• 金额: $ 40.55万
• 依托单位: HARVARD UNIVERSITY
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8535279
• 关键词: Affect; Behavior; Binding; Biological Markers; Biomedical Research; Biotin; Cell Culture Techniques; Cell Line; Cells; Chromatin; Color; Communities; DNA Methylation; Data; Defect; Development; Developmental Biology; Ectopic Expression; Embryo; Engineering; Epigenetic Process; Fibroblasts; Fluorescence-Activated Cell Sorting; Gene Expression Profile; Genome; Heterogeneity; Human; Immunoprecipitation; Instruction; Knock-in Mouse; Label; Link; Maintenance; Maps; Mass Spectrum Analysis; Methods; Modification; Mus; Pharmacology; Phenotype; Play; Pluripotent Stem Cells; Population; Principal Investigator; Process; Property; Proteins; Regenerative Medicine; Regulation; Regulatory Pathway; Reporter; Reporting; Research; Role; Signal Pathway; Somatic Cell; Stem cells; Testing; Transcriptional Activation; Variant; base; c-Myc Staining Method; c-myc Genes; cell type; chromatin remodeling; clinical application; embryonic stem cell; health care delivery; histone modification; induced pluripotent stem cell; insight; interest; pluripotency; programs; stem; tool; transcription factor

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.

发育生物学和生物医学界长期以来对理解 具有相同基因组的细胞如何建立并维持显着不同的行为和基因表达 模式。 尝试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素 鉴定出单个转录因子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编程为多能性 状态。转录因子的异位表达可能是一种通用且实用的方法 任意细胞状态之间的重编程。然而，从我们的初步研究中我们也知道 这些因素本身无法诱导完全转化，我们发现存在很大的异质性和 一旦建立多能状态，就会发生变异。 在这里，我们的目标是极大地增进我们对重编程和多能性本身的一般理解。 为了实现这一点，我们首先要利用重编程过程的方向性， 确定细胞上下文相关的重塑能力（目标1），然后生成多维数据 描述多能群体（目标 2），并最终构建了使我们能够进一步表征的工具 多能细胞培养物中存在的亚群（目标 3）。拟议的研究提供了一个机会 显着改变和加速细胞重编程在基础、转化和临床方面的效用 应用程序。 相关性（参见说明）： 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 影响细胞状态转变（体细胞到多能）以及随后的维持 多能状态。更好地理解多能性的建立和维持可以 改变生物医学研究和医疗保健服务。