Investigating the pathomechanisms underlying Charcot-Marie-Tooth Disease

研究腓骨肌萎缩症的病理机制

• 批准号: 10541701
• 负责人: Julia Alexis Jones
• 金额: $ 3.48万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-07-01 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10541701
• 关键词: Address; Affect; Amino Acids; Amino Acyl-tRNA Synthetases; Antioxidants; Auditory system; Axon; Binding Sites; Biological Assay; Brain Stem; Cell Line; Cell Nucleus; Cell physiology; Cells; Charcot-Marie-Tooth Disease; Clinical; Cochlea; Cytosol; DNA Damage; Defect; Development; Disease; Educational process of instructing; Educational workshop; Enzymes; Epithelial; Exhibits; Family; Family member; Fibroblasts; Gene Expression; Gene Family; Genes; Genetic Transcription; Goals; HSPB1 gene; Hereditary Motor and Sensory Neuropathy Type I; Hereditary Motor and Sensory-Neuropathy Type II; Hindlimb; Human; Image; Impaired cognition; Inherited; Investigation; Link; Longitudinal Studies; Maps; Metabolism; Midbrain structure; Mitochondria; Modeling; Molecular; Molecular Profiling; Molecular Target; Morphology; Motor; Motor Neurons; Mus; Mutate; Mutation; Myelin Sheath; Neuroglia; Neurologic Effect; Neurons; Neurosciences; Nuclear; Nuclear Localization Signal; Organelles; Organism; Oxidative Stress; PMP22 gene; Pathogenesis; Pathogenicity; Pathology; Pathway interactions; Patients; Peripheral Nervous System Diseases; Persons; Phase; Phenotype; Physiological; Physiology; Play; Production; Property; Protein Biosynthesis; Proteins; Research; Research Project Grants; Role; Schwann Cells; Sensory; Signal Pathway; Specificity; Stress; Technical Expertise; Therapeutic; Tissue Sample; Transfer RNA; Transfer RNA Aminoacylation; Tyrosine-tRNA Ligase; United States; Writing; auditory pathway; axon injury; base; biological adaptation to stress; career; cell type; druggable target; fly; hearing impairment; in vivo; induced pluripotent stem cell; insight; kidney cell; knock-down; member; mitochondrial dysfunction; mouse model; mutant; response; sex; spiral ganglion; symposium; therapeutic development; transcriptome sequencing

PROJECT SUMMARY Charcot-Marie-Tooth (CMT) disease is a genetically and clinically heterogeneous group of inherited peripheral neuropathies that is characterized by damage to long motor and sensory axons. The overall goal of this project is to identify the molecular and cellular processes that are shared between different CMT genes to help address a common target for treatment for this currently incurable disease. Our lab extensively studies aminoacyl-tRNA synthetases (aaRS), the largest gene family involved in CMT, and discovered the nuclear function of one aaRS, TyrRS, was shown to be associated to CMT pathogenesis while acting as a transcriptional regulator under oxidative stress. Aim 1 sets out to understand how the nuclear aaRS function in oxidative stress response plays a role in mouse physiology and if it is the commonality between CMT-linked aaRS. Findings so far suggest nuclear TyrRS in mice leads to an increased metabolism and disruption in the auditory system. This proposal sets out to perform a longitudinal study to determine if the hearing loss is degenerative and where in the auditory pathway there is a defect (e.g., sensory epithelium, spiral ganglion, brainstem, midbrain) while also examining the molecular targets responsible using RNA-Sequencing on tissue samples. Additionally, we uncovered that CMT-linked aaRS share a similar nuclear localization response upon oxidative stress treatment in neuronal cells only. Aim 1 also sets out to investigate this apparent cell type specificity, examine more cell lines, and evaluate the factor responsible for the translocation property. More broadly, I want to understand the connection between multiple CMT genes and subtypes to have a holistic view of the cellular organelles and signaling pathways underlying CMT pathomechanisms. Therefore, in Aim 2 as a postdoctoral trainee I will study a range of CMT proteins across subtypes of varying phenotypes. Patient derived, induced pluripotent stem cells will be differentiated into two relevant CMT cell types for each mutant: motor neurons and Schwann cells. Because there is evidence suggesting mitochondrial defects and oxidative stress in some CMT subtypes, I will specifically investigate mitochondrial morphology, dynamics, and function using imaging and cell-based assays, as well as, exploring the levels of oxidate stress, stress-related damage, and potential pathways affected in CMT in a cell type specific manner. During this proposal, activities such as teaching, writing workshops, mastery of technical skills, and conference participation will be emphasized to equip me with the professional development needed for an independent career in neuroscience.

项目概要 腓骨肌萎缩症 (CMT) 是一组遗传和临床异质性的疾病 遗传性周围神经病，其特征是长运动和感觉受损 轴突。该项目的总体目标是确定分子和细胞过程 不同 CMT 基因之间共享，以帮助解决该疾病的共同治疗目标 目前为不治之症。我们的实验室广泛研究氨酰基-tRNA 合成酶 (aaRS)， 参与 CMT 的最大基因家族，并发现了一个 aaRS 的核功能， TyrRS，被证明与 CMT 发病机制相关，同时充当转录因子 氧化应激下的调节剂。目标 1 旨在了解核 aaRS 如何在 氧化应激反应在小鼠生理学中发挥着重要作用，如果它是两者之间的共性 CMT 连接的 aaRS。迄今为止的研究结果表明，小鼠体内的核 TyrRS 会导致 新陈代谢和听觉系统的破坏。该提案旨在执行一项 纵向研究以确定听力损失是否是退行性的以及听觉的哪个部位 通路存在缺陷（例如感觉上皮、螺旋神经节、脑干、中脑） 还使用组织样本上的 RNA 测序来检查负责的分子靶点。 此外，我们发现 CMT 连接的 aaRS 具有相似的核定位反应 仅在神经元细胞中进行氧化应激处理。目标 1 也着手对此进行调查 明显的细胞类型特异性，检查更多细胞系，并评估负责的因素 易位性质。更广泛地说，我想了解多个之间的联系 CMT 基因和亚型可全面了解细胞器和信号传导 CMT 病理机制的潜在途径。因此，在目标2中，作为博士后实习生我将 研究一系列跨不同表型亚型的 CMT 蛋白。患者衍生、诱导 对于每个突变体，多能干细胞将分化为两种相关的 CMT 细胞类型： 运动神经元和雪旺细胞。因为有证据表明线粒体缺陷 和某些 CMT 亚型中的氧化应激，我将专门研究线粒体 使用成像和基于细胞的检测来研究形态、动力学和功能，以及探索 氧化应激水平、应激相关损伤以及 CMT 中受影响的潜在途径 细胞类型特定方式。在此提案期间，诸如教学、写作研讨会等活动， 将强调掌握技术技能和参加会议，以使我具备 神经科学领域独立职业所需的专业发展。