权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Mechanisms of disease pathogenesis in neurofilament linked Charcot-Marie-Tooth di

神经丝相关夏科-马里-图思病的发病机制

基本信息

批准号：
7565896
负责人：
MICHAEL L GARCIA
金额：
$ 7.23万
依托单位：
UNIVERSITY OF MISSOURI-COLUMBIA
依托单位国家：
美国
项目类别：
财政年份：
2008
资助国家：
美国
起止时间：
2008-02-15 至 2010-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7565896
关键词：
Address Adult Affect American Amyotrophic Lateral Sclerosis Animal Model Appearance Arginine Axon Axonal Neuropathy Biological Assay Cell Culture Techniques Charcot-Marie-Tooth Disease Classification Cytoskeletal Proteins Development Disease Disease model Electron Microscopy Experimental Designs Functional disorder Gait Gene Targeting Genes Glutamates Hoarseness Human Infant Inherited Knock-in Mouse Larynx Lead Light Link Lysine Mediating Missense Mutation Monitor Motor Mus Muscle Mutant Strains Mice Mutate Mutation Nerve Neural Conduction Neurons Organism Pathogenesis Pathology Patients Perception Peripheral Nerves Peripheral Nervous System Diseases Phenotype Phosphorylation Proline Radial Relative (related person)Reporting Schwann Cells Sensory Series Severities Severity of illness Site Symptoms System Tertiary Protein Structure Therapeutic Intervention Time Tooth structure Walking Weight Withdrawal afferent nerve axon growth axonopathy cell type foot gene replacement genome wide association study insight interest mutant myelination neurofilament neuronal cell body neurotoxicity novel public health relevance respiratory sciatic nerve therapy development wasting

项目摘要

DESCRIPTION (provided by applicant): Jean Martin Charcot, Pierre Marie and Howard Henry Tooth characterized Charcot-Marie-Tooth (CMT) in 1886. Today, CMT is the most common inherited disease of the peripheral nervous system, affecting approximately 150,000 Americans. Four major forms of CMT are now recognized (referred to as CMT 1-4). The major differences between forms are the linked gene and the primary cell type affected (Schwann cells versus nerves). Charcot-Marie-Tooth type 2E (CMT2E) is a sub-type of CMT2. CMT2E is an autosomal dominant disorder that affects peripheral nerve axons. A series of recent reports linked 16 mutations in neurofilament light (NF-L) to CMT2E. NF-L, with CMT2E linked mutations, expressed in cell culture disrupts neurofilament organization and transport. Additionally, mutant NF-L functions in a dominant manner in primary neuronal cultures. Interestingly, complete loss of all axonal neurofilaments, through targeted deletion of NF-L in mouse, does not result in overt pathology. Therefore, little is known about the mechanism(s) involved in the pathogenesis of NF-L linked CMT2E. Our objective is to develop two animal models of CMT2E so that we can analyze disease pathogenesis in NF-L linked CMT. We will achieve this by developing two independent lines of gene knock-in mice. One line of mice will express NF-L with proline 8 mutated to arginine, and the other will express NF-L with glutamate 397 mutated to lysine. We will analyze these mice for pathological changes at both the cellular and organism level. At the cellular level, we will look for alterations in neurofilament accumulation and organization in axons, alterations in radial axonal growth and alterations in myelination. We will, also, monitor the mice for the appearance of CMT-like symptoms. Specifically, we will analyze muscle wasting, gait, thermal perception and nerve conduction velocities. Generating two lines of gene-targeted mice will address important questions relevant to therapy development. For example, as our proposed mutations are in distinct functional domains of the protein, do these mutations result in disease through different mechanisms? Do different NF-L mutations result in variable onset or severity of disease? Additionally, we will analyze cellular phenotypes independently in motor and sensory axons to determine if both neuronal types are equally vulnerable to expression of mutant NF-L. The use of a time course will allow us to identify early changes associated with expressing mutant NF-L. Generating and analyzing animal models of CMT2E is the first key step in identifying novel sites and strategies for therapeutic intervention. PUBLIC HEALTH RELEVANCE: Charcot-Marie-Tooth (CMT) is the most common inherited disease of the peripheral nervous system affecting approximately 150,000 Americans with severe cases of CMT2 presenting with respiratory dysfunction in infants and with laryngeal weakness, hoarseness and respiratory difficulties in adults. Mutations in a cytoskeletal protein, neurofilament light (NF-L), have been linked to CMT2E. We are interested in developing animal models of CMT2E to determine how mutations in NF-L lead to the development of disease and to identify novel sites and strategies for therapeutic intervention.

描述（由申请人提供）：Jean Martin Charcot、Pierre Marie 和 Howard Henry Tooth 在 1886 年描述了 Charcot-Marie-Tooth (CMT) 的特征。如今，CMT 是周围神经系统最常见的遗传性疾病，影响大约 150,000 名美国人。现已认可 CMT 的四种主要形式（称为 CMT 1-4）。不同形式之间的主要区别在于相关基因和受影响的主要细胞类型（雪旺细胞与神经）。腓骨肌萎缩症 2E 型 (CMT2E) 是 CMT2 的一个亚型。 CMT2E 是一种常染色体显性遗传疾病，影响周围神经轴突。最近的一系列报告将神经丝光 (NF-L) 的 16 个突变与 CMT2E 联系起来。细胞培养物中表达的具有 CMT2E 相关突变的 NF-L 会破坏神经丝组织和运输。此外，突变型 NF-L 在原代神经元培养物中以主导方式发挥作用。有趣的是，通过靶向删除小鼠中的 NF-L，所有轴突神经丝的完全丧失并不会导致明显的病理学。因此，人们对 NF-L 相关 CMT2E 发病机制所涉及的机制知之甚少。我们的目标是开发两种 CMT2E 动物模型，以便我们能够分析 NF-L 相关 CMT 的疾病发病机制。我们将通过开发两个独立的基因敲入小鼠品系来实现这一目标。一组小鼠将表达脯氨酸8突变为精氨酸的NF-L，另一组小鼠将表达谷氨酸397突变为赖氨酸的NF-L。我们将在细胞和有机体水平上分析这些小鼠的病理变化。在细胞水平上，我们将寻找轴突中神经丝积累和组织的改变、径向轴突生长的改变和髓鞘形成的改变。我们还将监测小鼠是否出现 CMT 样症状。具体来说，我们将分析肌肉消耗、步态、热感知和神经传导速度。产生两个基因靶向小鼠品系将解决与治疗开发相关的重要问题。例如，由于我们提出的突变位于蛋白质的不同功能域中，这些突变是否通过不同的机制导致疾病？不同的 NF-L 突变是否会导致不同的疾病发作或严重程度？此外，我们将独立分析运动和感觉轴突的细胞表型，以确定这两种神经元类型是否同样容易受到突变体 NF-L 表达的影响。时间进程的使用将使我们能够识别与表达突变型 NF-L 相关的早期变化。生成和分析 CMT2E 动物模型是确定新位点和治疗干预策略的关键第一步。公共卫生相关性：腓骨肌萎缩症 (CMT) 是最常见的周围神经系统遗传性疾病，影响约 150,000 名美国人，严重的 CMT2 病例表现为婴儿呼吸功能障碍，成人表现为喉部无力、声音嘶哑和呼吸困难。细胞骨架蛋白神经丝光 (NF-L) 的突变与 CMT2E 相关。我们有兴趣开发 CMT2E 动物模型，以确定 NF-L 突变如何导致疾病的发展，并确定治疗干预的新位点和策略。