A Drosophila model of motor neuron disease using mutations in P150 / Dynactin.

描述（由申请人提供）：肌萎缩侧索硬化症（ALS）是一种致死性运动神经元神经退行性疾病，病因不明，治疗无效。我们对ALS发病机制的认识不足，部分原因是缺乏简单的运动神经元变性动物模型。候选人医学博士托马斯·劳埃德博士建议使用强大的遗传生物果蝇来表征和利用运动神经元疾病的新模型。新出现的数据表明，轴突运输或其他囊泡运输事件的缺陷可能是这种疾病的主要原因。为了研究囊泡转运在运动神经元疾病中的作用，我们将一种存在于罕见的ALS家族形式中的dynactin的P150亚基突变引入果蝇。表达突变P150的果蝇具有几种ALS的表型，包括突变蛋白的聚集、轴突运输缺陷、成年发病、进行性麻痹和早期死亡。该提案的目标是进一步描述运动神经元疾病的简单遗传模型，然后使用它来筛选运动神经元变性的遗传抑制因子。目的1研究疾病相关的P150突变对果蝇体内外囊泡转运的影响，以验证这些突变破坏特定囊泡转运过程的假设。目的2将检验疾病相关P150突变导致运动神经元和神经肌肉接头病理学类似于ALS患者的假设。目的3将首先表征突变型P150的已鉴定抑制剂对这些运动神经元表型的影响。然后，我们将筛选果蝇基因组中突变型P150的其他遗传修饰剂，希望确定对疾病发病机制至关重要的新基因，这是一种在小鼠模型中不可行的强大方法。确定的遗传相互作用是潜在的药物靶点，因此该提案的未来方向是验证ALS小鼠模型中确定的相互作用基因。拟议的研究将在果蝇和小鼠运动神经元连接专家Alex Kolodkin的实验室进行，并由Robert Packard Center for ALS Research主任Jeff罗斯坦指导。约翰霍普金斯大学的神经科学和神经病学系为学术神经学家的发展提供了一个特殊的环境。该补助金提供的培训和指导将为劳埃德博士提供成为一名成功的独立医生-科学家所需的专业知识和工具，他将致力于为ALS确定新的治疗方法。 相关性：ALS治疗的发展受到缺乏这种疾病的简单动物模型的阻碍。该提案将描述一种新的ALS果蝇模型，以帮助了解ALS的遗传原因，并帮助寻找这种毁灭性疾病的新药靶点。

DESCRIPTION (provided by applicant): Amyotrophic Lateral Sclerosis (ALS) is a fatal neurodegenerative disease of motor neurons without a known cause or effective treatment. Our poor understanding of ALS pathogenesis is partly due to a lack of simple animal models of motor neuron degeneration. The candidate, Thomas Lloyd M.D., Ph.D. proposes to characterize and utilize a novel model of motor neuron disease using the powerful genetic organism Drosophila melanogaster. Emerging data suggest that defects in axonal transport or other vesicle trafficking events may be a primary cause of this disease. To investigate the role of vesicle transport in motor neuron disease, we have introduced a mutation in the P150 subunit of dynactin into Drosophila that is present in a rare familial form of ALS. Flies expressing mutant P150 have several phenotypes reminiscent of ALS including aggregates of mutant protein, defects in axonal transport, and adult-onset, progressive paralysis and early death. The goal of this proposal is to further characterize this simple genetic model of motor neuron disease, and then to use it to screen for genetic suppressors of motor neuron degeneration. Aim 1 will investigate the effects of disease-associated P150 mutations on vesicle transport in Drosophila in vitro and in vivo to test the hypothesis that these mutations disrupt specific vesicle transport processes. Aim 2 will test the hypothesis that disease-associated P150 mutations cause motor neuron and neuromuscular junction pathology resembling that seen in ALS patients. Aim 3 will first characterize the effect of an identified suppressor of mutant P150 on these motor neuron phenotypes. We will then screen the Drosophila genome for additional genetic modifiers of mutant P150 in hopes of identifying novel genes critical to disease pathogenesis, a powerful approach not feasible in mouse models. Identified genetic interactors are potential drug targets, so a future direction of this proposal is to validate identified interacting genes in mouse models of ALS. The studies proposed will be carried out in the laboratory of Alex Kolodkin, an expert in motor neuron connectivity in Drosophila and mice, with mentorship from Jeff Rothstein, Director of the Robert Packard Center for ALS Research. The neuroscience and neurology departments at Johns Hopkins provide an exceptional environment for the development of academic neurologists. The training and mentorship provided by this grant will give Dr. Lloyd the expertise and tools needed to become a successful, independent physician-scientist who will devote his career to identifying new treatments for ALS. RELEVANCE: The development of treatments for ALS is hindered by the lack of simple animal models of this disease. This proposal will characterize a new fruitfly model of ALS to help understand the genetic causes of ALS and to help find new drugs targets for this devastating disease.