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.

项目摘要 Charcot-Marie-Tooth（CMT）疾病是遗传和临床上异质的一组 遗传性周围神经病的特征是对长电动机和感觉损害 轴突。该项目的总体目标是确定分子和细胞过程 在不同的CMT基因之间共享，以帮助解决这一治疗的共同靶标 目前无法治愈的疾病。我们的实验室广泛研究氨基酰基-TRNA合酶（AARS），， 参与CMT的最大基因家族，发现了一个AAR的核功能， Tyrrs被证明与CMT发病机理有关，同时充当转录 调节剂在氧化应激下。 AIM 1开始了解核AAR的功能 氧化应激反应在小鼠生理学中起作用，如果是 CMT链接AARS。到目前为止的发现表明，小鼠的核pyrrs导致增加 听觉系统中的代谢和破坏。该提议开始执行 纵向研究以确定听力损失是否退化以及在听觉中的位置 途径有一个缺陷（例如感觉上皮，螺旋神经节，脑干，中脑） 还检查使用组织样品上的RNA测序负责的分子靶标。 此外，我们发现CMT链接的AAR共享类似的核定位响应 仅在神经元细胞中进行氧化应激治疗。 AIM 1还开始调查此事 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 转运属性。更广泛地，我想了解多个 CMT基因和亚型具有细胞细胞器和信号传导的整体视图 CMT病理机理的基础途径。因此，在AIM 2作为博士后学员，我将 研究跨不同表型亚型的一系列CMT蛋白。患者得出，诱导 多能干细胞将分为每种突变体的两种相关的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亚型的氧化应激，我将特别研究线粒体 使用成像和基于细胞的测定法以及探索的形态，动力学和功能 牛的应力水平，与应力相关的损伤以及在CMT中影响的潜在途径 细胞类型特定的方式。在此提案中，教学，写研讨会等活动， 掌握技术技能和会议参与将被强调为我配备 神经科学领域独立职业所需的专业发展。