Alpha-Tubulin Mutation and Motor Neuron Disease

α-微管蛋白突变和运动神经元疾病

• 批准号: 9297882
• 负责人: Mahmoud Kiaei
• 金额: $ 7.45万
• 依托单位: UNIV OF ARKANSAS FOR MED SCIS
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-03-01 至 2019-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9297882
• 关键词: Actins; Adult; Affect; American; Amyotrophic Lateral Sclerosis; Animal Model; Area; Axon; Back; Biological Assay; Body Weight decreased; Brain; C9ORF72; Cells; Code; Communities; Cytoskeleton; Defect; Development; Dimerization; Disease; Dynein ATPase; Familial Amyotrophic Lateral Sclerosis; Family; Functional disorder; Gait; Genes; Genome; Goals; Human; In Vitro; Inflammation; Investigation; Kinesin; Lead; Length; Limb Tremors; Link; Microtubule Polymerization; Microtubules; Mitochondria; Modeling; Monitor; Motor; Motor Neuron Disease; Motor Neurons; Mus; Muscle; Muscle Weakness; Muscular Atrophy; Mutant Strains Mice; Mutate; Mutation; Nerve Degeneration; Neurites; Neuroglia; Neuromuscular Junction; Neurons; Oxidative Stress; Pathogenesis; Pathogenicity; Pathologic; Pathology; Patients; Performance; Phenotype; Polymers; Process; Production; Proteins; Research; Role; Severities; Signs and Symptoms; Skeletal Muscle; Spinal Cord; Stress Fibers; Structure; Tail; Testing; Therapeutic; Toxic effect; Transgenes; Transgenic Mice; Tubulin; Ursidae Family; Work; alpha Tubulin; animal model development; anterograde transport; axon injury; base; cohort; dimer; dynactin; effective therapy; exome sequencing; gene discovery; glial activation; in vivo; interest; mitochondrial dysfunction; motor deficit; motor neuron degeneration; mouse model; mutant; nervous system disorder; neuromechanism; neuromuscular; neuron loss; neurotoxicity; novel; overexpression; prevent; protein expression; reduce symptoms; retrograde transport; superoxide dismutase 1; therapeutic development; therapeutic evaluation; tool; trend

PROJECT SUMMARY/ABSTRACT Amyotrophic lateral sclerosis (ALS) or Lou Gehrig's disease, is a fatal disease resulting from progressive degeneration of motor neurons and affects ~30,000 American each year. Currently, the mechanisms responsible for motor neuron death in ALS are not fully understood and there is no effective therapy to cure or slow the progression of this inexorable disease. The recent discovery of TUBA4A mutations in familial ALS patients added a new gene with implication in cytoskeleton dynamics in motor neurons. We became highly interested to investigate the role of mutant -tubulin in the cytoskeleton dynamics and defects in ALS. Toward that end, we have planned to generate novel transgenic mice overexpressing mutant human wildtype and mutant -tubulin. Our long-term goal is to understand the mutant -tubulin neurotoxicity in motor neuron degeneration and develop strategy to block its toxicity. Our immediate goal is to construct, develop a novel animal model that bears a mutation identical to that found in multiple families with ALS. We hypothesize that the overexpression of mutant -tubulin protein in mice will result in development of cardinal phenotypes and pathologies that resemble those in ALS patients. The first aim is to generate -tubulin transgenic mice and assess for progressive motor deficits and phenotypes resembling ALS. Adult mice will be studied for signs and symptoms that attributes to functional motor neuron degeneration, correlating the severity of the dysfunction to TUBA4A transgene copy number, protein expression level, and protein activity. The second aim is to characterize hTUBA4AR320C and hTUBA4AWT transgenic mice and compare neuromuscular pathologies with those of SOD1G93A and our recent hPFN1G118V transgenic mice. Brain, spinal cord and skeletal muscle tissues will be studied to determine pathological anomalies including damage to neuronal loss, axonal damage, production of oxidative stress, and glial activation. Microtubule polymerization and a-tubulin activity assays will be performed to determine if the R320C mutation disrupts the structure of the microtubule. The impact of this work could be highly significant for its potential in creating a new animal model for research areas complementary to existing ALS research. This study will provide a new tool to determine the central importance of mutant -tubulin as a basic mechanism that may contribute to the neurodegeneration found in ALS patients. We expect that this mouse model may also facilitate development of therapeutic strategies for ALS. Thus, the results are expected to have a significant impact because our model may provide new targets for discovery of basic mechanisms and therapies to prevent or stop progression of ALS-associated neurodegeneration.

项目总结/摘要 肌萎缩侧索硬化症（ALS）或卢伽雷氏病（Lou Gehrig's disease）是一种由进行性肌萎缩性侧索硬化（ALS）引起的致命疾病。 运动神经元退化，每年影响约30，000美国人。目前，这些机制 导致ALS中运动神经元死亡的原因尚未完全了解，并且没有有效的治疗方法来治愈或治疗ALS。 减缓这种不可阻挡的疾病的发展。家族性ALS中TUBA 4A突变的最新发现 患者增加了一个新的基因，暗示运动神经元的细胞骨架动力学。我们变得高度 感兴趣的是研究突变的微管蛋白在ALS细胞骨架动力学和缺陷中的作用。朝向 为此，我们计划产生新的过表达突变的人野生型的转基因小鼠， 突变型β-微管蛋白。我们的长期目标是了解运动神经元中的突变微管蛋白神经毒性 退化和发展战略，以阻止其毒性。我们的近期目标是构建、发展一部小说 携带与多个ALS家族中发现的突变相同的突变的动物模型。我们假设 突变型β-微管蛋白在小鼠中过表达将导致主要表型的发展， 类似于ALS患者的病理。第一个目的是产生β-微管蛋白转基因小鼠， 评估进行性运动缺陷和类似ALS的表型。将研究成年小鼠的体征， 归因于功能性运动神经元变性的症状，将功能障碍的严重程度与 TUBA 4A转基因拷贝数、蛋白表达水平和蛋白活性。 第二个目的是表征hTUBA 4AR 320 C和hTUBA 4AWT转基因小鼠，并比较 神经肌肉病理学与SOD 1G 93 A和我们最近的hPFN 1G 118 V转基因小鼠的神经肌肉病理学相似。脑、脊髓 将研究脊髓和骨骼肌组织，以确定病理异常，包括 神经元损失、轴突损伤、氧化应激的产生和神经胶质活化。微管聚合 并且将进行α-微管蛋白活性测定以确定R320 C突变是否破坏微管蛋白的结构。 微管 这项工作的影响可能是非常重要的，因为它有可能创造一种新的动物模型， 与现有ALS研究互补的研究领域。该研究将为确定 突变型β-微管蛋白作为可能导致神经变性的基本机制的重要性 在ALS患者中发现。我们希望这种小鼠模型也可以促进治疗性药物的开发。 ALS的策略因此，预计结果将产生重大影响，因为我们的模型可以提供 发现基本机制和治疗的新靶点，以预防或阻止ALS相关疾病的进展。 神经变性