Investigating the pathomechanisms underlying Charcot-Marie-Tooth Disease

项目摘要 腓骨肌萎缩症（CMT）是一组遗传和临床异质性的 以长运动和感觉受损为特征的遗传性周围神经病 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，以帮助解决治疗这种疾病的共同目标。 目前无法治愈的疾病。我们实验室广泛研究氨酰-tRNA合成酶（阿尔斯）， 最大的基因家族参与CMT，并发现了一个阿尔斯的核功能， TyrRS与CMT发病机制相关，同时作为转录调节因子发挥作用。 氧化应激下的调节剂。目的1旨在了解核阿尔斯如何在 氧化应激反应在小鼠生理学中起作用，如果它是 CMT相关阿尔斯。到目前为止的研究结果表明，小鼠的核TyrRS导致了小鼠细胞内TyrRS水平的增加。 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，以及在听觉 存在缺陷的路径（例如，感觉上皮、螺旋神经节、脑干、中脑）， 还使用RNA测序对组织样本进行分子靶点检测。 此外，我们发现CMT连锁的阿尔斯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。Aim 1也开始研究这一点 明显的细胞类型特异性，检查更多的细胞系，并评估负责的因素 易位性质。更广泛地说，我想了解多个 CMT基因和亚型对细胞器和信号传导有一个整体的看法 CMT病理机制的潜在途径。因此，在目标2中，作为博士后培训生，我将 研究一系列不同表型亚型的CMT蛋白。患者衍生，诱导 对于每种突变体，多能干细胞将分化成两种相关的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 和氧化应激在一些CMT亚型，我将专门研究线粒体 形态学、动力学和功能，使用成像和基于细胞的测定，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， 细胞类型特异性方式。在此期间，活动，如教学，写作研讨会， 掌握技术技能，并参加会议将被强调，以装备我与 专业发展需要在神经科学的独立职业生涯。

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.