Motor neuron selector genes and mechanism of their action

运动神经元选择基因及其作用机制

• 批准号: 8537518
• 负责人: Hynek Wichterle
• 金额: $ 53.88万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-09-01 至 2017-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8537518
• 关键词: Address; Adopted; Algorithms; Axon; Binding; Biochemical; Biological Assay; Cell physiology; Cells; ChIP-seq; Chromatin; Chromatin Structure; Clinical; Competence; Computer Analysis; DNA; DNA Binding; DNA-Protein Interaction; Development; Developmental Process; Disease model; Enhancers; Event; Failure; Gene Expression; Generations; Genes; Genetic Transcription; Genomics; Individual; Maps; Methods; Molecular; Molecular Conformation; Motor Neuron Disease; Motor Neurons; Muscle; Mutation Analysis; Ontology; Output; Pattern; Pluripotent Stem Cells; Population; Process; Recruitment Activity; Refractory; Regulation; Reporter; Signal Transduction; Source; Specificity; Spinal Cord; Staging; System; Testing; Transplantation; Universities; cell motility; cell type; cholinergic; clinical application; clinically relevant; design; drug discovery; embryonic stem cell; genome-wide; in vitro Assay; in vivo; insight; network models; novel; progenitor; programs; stem cell biology; synaptogenesis; syntax; transcription factor

DESCRIPTION (provided by applicant): Transcriptional programming of cell identity is gaining importance at both basic developmental and clinical level. While the phenomenology of cell programming and reprogramming by forced expression of transcription factors is well described, the mechanism of action of programming factors or the sequence of regulatory events resulting in a cell adopting a new identity are largely unknown. We propose to combine the strengths of stem cell biology with genomic and computational approaches to characterize the process of transcriptional programming of motor neuron (MN) identity. We developed efficient methods for the induction of MN identity in differentiating embryonic stem cells (ESCs) by the expression of defined transcription factors. Using the system we will combine biochemical, genomic and computational analysis to address following questions: i) whether programming factors directly regulate terminal motor neuron effector genes or initiate a cascade of intermediate transcription programs; ii) whether recruitment of programming factors to DNA is cooperative and which factors determine the specificity of DNA binding; iii) whether identified MN enhancers are inaccessible for programming factor binding in cell types refractory to MN programming; iv) whether identified secondary binding motifs for Onecut and Ebf transcription factors contribute to productive regulation of MN specific gene expression; v) whether additional secondary motifs recruit ancillary transcription factors to NIL bound enhancers that contribute to productive regulation of MN specific gene expression. Together these studies will provide fundamental insight into the developmental processes underlying specification of defined cell identity and will provide novel and efficient source of motor neurons for disease modeling, study and drug discovery.

描述（由申请人提供）：细胞身份的转录编程在基础发育和临床水平上都变得越来越重要。虽然通过转录因子的强制表达进行细胞编程和重编程的现象学已得到很好的描述，但编程因子的作用机制或导致细胞采用新身份的调节事件序列在很大程度上是未知的。我们建议将干细胞生物学的优势与基因组和计算方法相结合，以表征运动神经元（MN）身份的转录编程过程。我们开发了通过表达特定转录因子来诱导分化胚胎干细胞 (ESC) 中 MN 身份的有效方法。使用该系统，我们将结合生化、基因组和计算分析来解决以下问题：i）编程因子是否直接调节末端运动神经元效应基因或启动一系列中间转录程序？ ii) 向 DNA 招募编程因子是否是合作性的以及哪些因素决定 DNA 结合的特异性； iii) 已识别的 MN 增强子是否无法在 MN 编程难治的细胞类型中与编程因子结合； iv) Onecut 和 Ebf 转录因子的二级结合基序是否有助于 MN 特异性基因表达的有效调控； v)是否有额外的二级基序将辅助转录因子募集到NIL结合的增强子中，从而有助于MN特异性基因表达的有效调节。这些研究将共同​​提供对特定细胞身份规范背后的发育过程的基本见解，并将 为疾病建模、研究和药物发现提供新颖有效的运动神经元来源。