Transcriptional Regulation of Phrenic Motor Neuron Identity

膈运动神经元身份的转录调节

• 批准号: 10593163
• 负责人: Ritesh KC
• 金额: $ 3.97万
• 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/10593163
• 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; Neuronal Differentiation; Neurons; Pathway interactions; Pattern; Process; Proteins; Reproducibility; Respiration; Respiration Disorders; Respiratory Diaphragm; Respiratory physiology; Rett Syndrome; Role; Site; Sleep Apnea Syndromes; Specific qualifier value; Specificity; Spinal Cord; Sudden infant death syndrome; Testing; Therapeutic; Transcriptional Regulation; cell type; cofactor; combinatorial; developmental disease; domain mapping; 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.

项目摘要 呼吸是地球生命的基本要素。在哺乳动物中，膈运动神经元（MN）形成单个运动神经元， 膈运动柱（Phrenic Motor Column或PMC），并以常规方式神经支配膈，以控制其运动。 收缩。膈神经的发育变化可能是呼吸功能障碍的原因， 在睡眠呼吸暂停和婴儿猝死综合征（SIDS）中观察到。尽管膈神经的重要作用， 建立膈神经MN同一性的分子决定因素还没有完全了解。我们以前的研究 已经表明Hox5（Hoxa5和Hoxc5）转录因子（TF）对于PMC的特化是关键的， 但是控制PMC特化的转录和调节机制还有待阐明。 破译Hox 5蛋白如何选择性控制其靶点对于了解膈神经至关重要 MN发展。Hox蛋白与选择辅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。在这个提议中，我将研究Hoxa5的功能 蛋白质及其与其他辅因子的相互作用，以确定PMC身份。 在目标1中，我将确定Hoxa 5在膈神经特异性表达过程中如何特异性调节其靶效应子。 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依赖性膈神经MN多样性。 我开发了一种综合方法，包括遗传模型，细胞和分子生物学 技术和高通量测序，以便在体内解决这些问题。阐明 膈运动神经元特性的转录调节的分子机制将使我们能够 确定潜在的治疗靶点，使我们更接近呼吸道疾病的有效治疗方法的发展。 功能障碍