INDUCTION AND MAINTENANCE OF PLURIPOTENT-PLASTIC STATE

多能塑性状态的诱导和维持

• 批准号: 6794033
• 负责人: ADAM S ASCH
• 金额: $ 38.14万
• 依托单位: WEILL MEDICAL COLL OF CORNELL UNIV
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-09-30 至 2006-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6794033
• 关键词: cell differentiation; genetic translation; high throughput technology; messenger RNA; pluripotent stem cells; tissue /cell culture

DESCRIPTION (provided by applicant): Recently, the traditional view that somatic stem cells remain faithful to their tissue of origin has been challenged. The capability of muscle-derived, and neural-derived cultures to produce cells capable of hematopoietic reconstitution following transplant, as well as the ability of hematopoietic cultures to give rise to muscle cells raises profound questions about the plasticity of somatic cells. The possibility that such plasticity might be exploited for therapeutic potential is compelling and makes a better understanding of its nature and the ways in which might be regulated of great interest. Plasticity, or the ability of cells that we think of as being lineage committed, to generate an alternative tissue or cell type must require the cells to reprogram and take an alternative developmental path. This must be true even of the stem cell populations within a tissue. An understanding of the basic mechanisms that define the pluripotent and plastic state somatic stem cells is the focus of this proposal. Our hypothesis is that the plasticity of stem cells a function of the translational repression of critical transcription factors present in the pluripotent stem cell. Plasticity, we also hypothesize, involves inhibition of expression of lineage defining transcripts already present within a cell resetting the developmental program along an alternative developmental pathway. To date, a proteomic approach to defining gene expression has not been undertaken. We have explored a novel and robust method for defining the change in translation upon the differentiation of stem cells. Using this approach we will be able to identify and study transcripts that are translationally repressed in the plastic/pluripotent state and gain an understanding of the basic mechanisms that underly plasticity and pluripotency. Specifically we will 1) identify critical translationally regulated mRNA transcripts and define an accurate picture of overall gene expression in the pluripotent cell by quantitating polysome-associated transcripts and their translational state using a rapid throughout screening assay in an in vitro model of stem cell plasticity; 2) define the mechanisms of translational regulation in transcripts critical to plasticity and pluripotency; and 3) define the role of translational regulation in modulating and maintaining pluripotency.

描述（由申请人提供）： 最近，传统的观点认为，体干细胞仍然忠实于 它们的组织来源受到了质疑 肌肉衍生的能力， 和神经衍生的培养物来产生能够造血的细胞 移植后的重建，以及造血的能力， 培养产生肌肉细胞提出了深刻的问题 体细胞的可塑性 这种可塑性可能是 开发治疗潜力是令人信服的， 了解它的性质和方式，其中可能受到管制的伟大 兴趣 可塑性，或者说细胞的能力， 为了生成替代组织或细胞类型，必须要求 细胞重新编程，并采取另一种发展道路。这必须 甚至对组织内的干细胞群体也是如此。 的理解 定义多能性和可塑性状态的基本机制 干细胞是这项建议的重点。 我们的假设是 干细胞的可塑性是一种翻译抑制功能， 多能干细胞中存在的关键转录因子。 可塑性，我们还假设，涉及抑制表达的谱系 定义已经存在于细胞内的转录本， 沿着另一条发展道路进行规划。 迄今为止，蛋白质组学 定义基因表达的方法还没有被采用。 我们有 探索了一种新的和强大的方法来定义翻译的变化， 干细胞的分化。 使用这种方法，我们将能够 识别和研究转录本，这些转录本在转录过程中受到抑制。 可塑性/多能性状态，并了解基本机制 这是可塑性和多能性的基础。 具体而言，我们将1）识别 关键的转录调控的mRNA转录，并定义一个准确的 通过定量获得多能细胞中总体基因表达的图片 多核糖体相关的转录本及其翻译状态， 在干细胞可塑性的体外模型中的整个筛选测定中; 2） 定义转录本中的翻译调控机制， 可塑性和多能性; 3）定义翻译的作用 在调节和维持多能性中的调节。