Exploring the Transcriptional Network regulating ES cell Pluripotency

探索调节 ES 细胞多能性的转录网络

• 批准号: 7392268
• 负责人: Sridhar Rao
• 金额: $ 13.31万
• 依托单位: DANA-FARBER CANCER INST
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-07-01 至 2012-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7392268
• 关键词: Affect; Affinity Chromatography; Area; Basic Science; Binding; Biological; Boston; Cancer Center; Cellular biology; Classification; Clinical; Clinical Medicine; Collaborations; Complex; Data; Development; Disease; Fostering; Gene Expression; Gene Targeting; Generations; Genes; Genetic Engineering; Genetic Transcription; Genome; In Vitro; Laboratories; Maintenance; Maps; Mass Spectrum Analysis; Medicine; Methods; Mus; Numbers; Pathway Analysis; Pathway interactions; Physicians; Play; Property; Proteins; Proteomics; Recording of previous events; Regenerative Medicine; Research; Research Personnel; Research Project Grants; Role; Role playing therapy; STAT3 gene; Scientist; Signal Transduction; Somatic Cell; Specificity; Stem cells; Systems Biology; Time; Tissues; Training; Transcriptional Regulation; Undifferentiated; Work; Yeasts; abstracting; base; career; embryonic stem cell; improved; insight; member; novel; pluripotency; prevent; programs; protein protein interaction; tool; transcription factor; yeast two hybrid system

DESCRIPTION (provided by applicant): Embryonic Stem cells provide both a unique biological tool for answering developmental questions, but also hold significant clinical promise in the area of regenerative medicine. Murine Embryonic Stem Cells (mESCs), which have existed for almost 25 years, are now being studied extensively for their ability to be maintained in vitro for extended periods of time in a pluripotent state (i.e. able to recreate all the tissues within a mouse). Understanding the mechanisms used to maintain pluripotency may allow for the reprogramming of somatic cells, which opens entirely new treatment options for a variety of diseases. This proposal involves a five-year training plan to foster a career in academic medicine, combining basic science with clinical medicine. This proposal utilizes a combination of stem-cell biology, proteomics, and systems biology to answer questions about mESC pluripotency. It will be performed within the Laboratory of Dr. Stuart Orkin, a leader in the field of stem cell biology who has trained numerous physician scientists. The research project will focus on the transcriptional mechanisms used to maintain mESCs in a pluripotent state. Recent work has shown that a small number of key transcription factors, Nanog, Oct4, and StatS appear to play a key role by regulating a large number of target genes. The mechanisms of transcriptional regulation, which are likely based upon key protein: protein interactions, remains to be elucidated. Initial work has revealed some of the key protein complexes formed in mESCs, although the binary interactions remain to be mapped out. In addition, how these protein: protein interactions occur on a genome-wide scale and affect target gene transcription are currently unknown. For this reason, a systematic yeast two-hybrid approach will be used to first confirm the binary interactions that are used to form these transcriptional complexes. Next, the key complexes formed by STATS will be identified using both new and established proteomics approaches. Lastly, to better understand the protein interactions critical for maintenance of mESCs, multiple yeast two-hybrid screens will be used to generate an "interactome", thereby facilitating a systems biology approach to the problem with significant network analysis. This proposal is ideally performed at the DFCI and CHB in Boston, given the long history of physician-scientist training, but also the unique collaboration with the Center for Cancer Systems Biology. (End of Abstract)

描述（由申请人提供）： 胚胎干细胞不仅为回答发育问题提供了独特的生物学工具，而且在再生医学领域也具有重要的临床前景。小鼠胚胎干细胞（mESC）已经存在了近25年，现在正在广泛研究它们在体外以多能状态长时间维持的能力（即能够重建小鼠体内的所有组织）。了解用于维持多能性的机制可能允许体细胞的重编程，这为各种疾病开辟了全新的治疗选择。该提案涉及一项五年培训计划，以促进学术医学的职业生涯，将基础科学与临床医学相结合。该提案利用干细胞生物学，蛋白质组学和系统生物学的组合来回答有关mESC多能性的问题。它将在Stuart Orkin博士的实验室内进行，Stuart Orkin博士是干细胞生物学领域的领导者，他培养了许多医生科学家。该研究项目将重点关注用于维持mESC多能状态的转录机制。最近的研究表明，少数关键转录因子Nanog、Oct 4和StatS似乎与转录因子的表达相关。 通过调控大量靶基因发挥关键作用。转录调控的机制，这可能是基于关键蛋白质：蛋白质相互作用，仍有待阐明。最初的工作已经揭示了一些在mESC中形成的关键蛋白质复合物，尽管二元相互作用仍有待确定。此外，这些蛋白质：蛋白质相互作用如何在全基因组范围内发生并影响靶基因转录目前尚不清楚。出于这个原因，将使用系统的酵母双杂交方法来首先确认用于形成这些转录复合物的二元相互作用。接下来，将使用新的和已建立的蛋白质组学方法来鉴定由STATS形成的关键复合物。最后，为了更好地理解对维持mESC至关重要的蛋白质相互作用，将使用多个酵母双杂交筛选来产生“相互作用组”，从而促进系统生物学方法来解决具有显著网络分析的问题。鉴于医生-科学家培训的悠久历史，以及与癌症系统生物学中心的独特合作，该提案在波士顿的DFCI和CHB进行了理想的执行。 (End摘要）