Transcriptional Regulation of Phrenic Motor Neuron Identity

膈运动神经元身份的转录调节

• 批准号: 10570824
• 负责人: Ritesh KC
• 金额: $ 3.95万
• 依托单位: CASE WESTERN RESERVE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-12-01 至 2023-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10570824
• 关键词: Address; Amino Acids; Area; Axon; Binding; Binding Sites; Biological Assay; Breathing; Cells; Cellular biology; Cervical spinal cord structure; Cessation of life; ChIP-seq; Chromatin; Co-Immunoprecipitations; DNA; DNA Binding; DNA Sequence; Development; Disease; Enhancers; Ensure; Environment; Excision; Family; Gene Expression; Genes; Genetic; Genetic Diseases; Genetic Models; Genetic Transcription; Global Change; HOX protein; High-Throughput Nucleotide Sequencing; Homeobox Genes; In Vitro; Intramuscular; Lateral; Lead; Life; Limb structure; Luciferases; Mammals; Maps; Mediating; Methodology; Molecular; Molecular Biology Techniques; Molecular Profiling; Motor; Motor Neurons; Muscular Dystrophies; Nervous system structure; Neurons; Pathway interactions; Pattern; Process; Proteins; Reproducibility; Respiration; Respiration Disorders; Respiratory Diaphragm; Respiratory physiology; Rett Syndrome; Role; Site; Sleep Apnea Syndromes; Specificity; Spinal Cord; Sudden infant death syndrome; Testing; Therapeutic; Transcriptional Regulation; cell type; cofactor; combinatorial; developmental disease; effective therapy; homeodomain; improved; in vivo; insight; motor neuron development; neural circuit; neuron loss; new therapeutic target; paralogous gene; programs; promoter; respiratory; single-cell RNA sequencing; spatiotemporal; therapeutic target; tissue oxygenation; transcription factor; transcription factor Oct-6

PROJECT SUMMARY Breathing is essential for terrestrial life. In mammals, phrenic motor neurons (MNs) form a single motor column (Phrenic Motor Column or PMC) and innervate the diaphragm in a stereotypical manner to control its contraction. Alterations in the development of phrenic MNs are likely a cause of respiratory dysfunction, as observed in sleep apneas and sudden infant death syndrome (SIDS). Despite the essential role of phrenic MNs, the molecular determinants that establish phrenic MN identity are not fully understood. Our previous studies have shown that Hox5 (Hoxa5 and Hoxc5) transcription factors (TFs) are critical for the specification of PMC, but the transcriptional and regulatory mechanisms that control PMC specification are yet to be elucidated. Deciphering how Hox5 proteins can selectively control their targets is fundamental in order to understand phrenic MN development. Cooperative binding of Hox proteins with select cofactors influences their target site selectivity. We have identified additional TFs that interact with Hoxa5. In this proposal I will investigate the function of Hoxa5 protein and its interaction with other cofactors in determining PMC identity. In Aim 1 I will determine how Hoxa5 specifically regulates its target effectors during the specification of phrenic MNs. In Aim 2 I will map the interaction between Hoxa5 and other cofactors and evaluate if this interaction is essential for the activation of putative phrenic enhancers and promoters. In Aim 3 I will define Hox5-dependent phrenic MN diversity. I have developed an integrative methodology encompassing genetic models, cell and molecular biology techniques, and high throughput sequencing in order to address these questions in vivo. Elucidating the molecular mechanisms underlying transcriptional regulation of phrenic motor neuron identity will allow us to identify potential therapeutic targets and bring us closer to the development of effective treatments for respiratory dysfunction.

项目概要 呼吸对于陆地生命至关重要。在哺乳动物中，膈运动神经元 (MN) 形成单个运动 柱（膈运动柱或 PMC）并以典型的方式支配隔膜以控制其 收缩。膈 MN 发育的改变可能是呼吸功能障碍的原因，因为 在睡眠呼吸暂停和婴儿猝死综合症（SIDS）中观察到。尽管膈 MN 发挥着重要作用， 建立膈 MN 身份的分子决定因素尚未完全了解。我们之前的研究 已经表明 Hox5（Hoxa5 和 Hoxc5）转录因子（TF）对于 PMC 的规范至关重要， 但控制 PMC 规范的转录和调控机制尚未阐明。 破译 Hox5 蛋白如何选择性控制其靶标对于理解膈神经至关重要 明尼苏达的发展。 Hox 蛋白与选定辅助因子的协同结合影响其靶位点选择性。 我们已经确定了与 Hoxa5 相互作用的其他 TF。在这个提案中我将研究 Hoxa5 的功能 蛋白质及其与其他辅助因子的相互作用在确定 PMC 身份中的作用。 在目标 1 中，我将确定 Hoxa5 在膈规范期间如何具体调节其目标效应器 MN。 在目标 2 中，我将绘制 Hoxa5 和其他辅因子之间的相互作用，并评估这种相互作用是否必要 用于激活假定的膈增强子和启动子。 在目标 3 中，我将定义 Hox5 依赖的膈 MN 多样性。 我开发了一种涵盖遗传模型、细胞和分子生物学的综合方法 技术和高通量测序，以在体内解决这些问题。阐明 膈运动神经元身份转录调控的分子机制将使我们能够 确定潜在的治疗靶点，使我们更接近开发有效的呼吸系统治疗方法 功能障碍。