Genetic repair of muscular degeneration associated with Duchenne muscular dystrophy

杜氏肌营养不良症相关肌肉变性的基因修复

• 批准号: 10439290
• 负责人: Martin F. Engelke
• 金额: $ 37.54万
• 依托单位: ILLINOIS STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-06-01 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10439290
• 关键词: Acute; Address; Affect; Afferent Neurons; Animal Model; Animals; Area; Award; Behavior; Behavioral; Biological Assay; Biological Models; Brain; Caenorhabditis elegans; Calcium; Calmodulin; Canis familiaris; Cell Line; Cell membrane; Cell physiology; Cessation of life; Chemicals; Complement; Coupling; DNA Sequence Alteration; Defect; Degenerative Disorder; Development; Disease; Disease Progression; Duchenne muscular dystrophy; Dystrophin; Embryonic Development; Exertion; Extracellular Matrix; Failure; Fishes; Functional disorder; Genes; Genetic; Grant; Growth; Homeostasis; Human; Immunohistochemistry; Impairment; Individual; Intervention; Larva; Lesion; Link; Longevity; Mediating; Mitochondria; Modeling; Molecular Target; Mus; Muscle; Muscle Cells; Muscle Development; Muscular Dystrophies; Mutation; Myoblasts; Myopathy; Necrosis; Nematoda; Nerve Degeneration; Nervous system structure; Neurologic; Neurons; Nociceptors; Orthologous Gene; Outcome; Pathogenesis; Pathology; Pathway interactions; Patients; Phenotype; Physical activity; Play; Preclinical Testing; Prevention; Process; Proteins; Public Health; RNA Interference; RNA interference screen; Regimen; Reporter; Role; Sensory; Severity of illness; Signal Pathway; Signal Transduction; Structure; System; Testing; Therapeutic; Titrations; United States National Institutes of Health; Validation; Work; advanced system; base; behavioral phenotyping; burden of illness; fly; high throughput screening; improved; insight; link protein; loss of function mutation; male; muscle degeneration; mutant; myogenesis; nervous system development; patient response; pre-clinical therapy; relating to nervous system; repaired; screening; side effect; therapeutic candidate; therapeutic target; treatment response

Project Summary/Abstract: Duchenne Muscular dystrophy (Dmd) is a lethal degenerative disease affecting 1 in 5,000 males. Dmd is caused by mutations in the gene encoding dystrophin, a highly conserved protein linking muscle cell membranes with the extracellular matrix and the contractile machinery within them. Dystrophin has structural and signaling functions. Loss of dystrophin is linked to muscular and neural degeneration. While traditional analyses of mice, worms and other animals modeling Dmd genetically, through loss-of-function mutations in the dystrophin gene, resulted in great advances, these systems have only produced relatively mild muscular and behavioral phenotypes. To date there is no cure for Dmd. To model the acute muscle degeneration observed in Dmd patients in a model system amenable to genetics we developed a fast and inexpensive nematode assay. Our assay elicits strong behavioral and cellular phenotypes in dystrophic (dys-1) nematodes to a degree not previously attained in other systems. During our previous award cycle, we improved our assay to allow automatization and medium to high throughput screening of candidate treatments. We went on to characterize many dystrophic phenotypes and found that they first arise during embryogenesis. We also identified the first neurological impairments in dystrophic worms, where the sensory function of ASH neurons is impaired. A suppressor mutant, and an RNA-interference screen both pointed to calmodulin as a therapeutic target. The first specific aim of this project is to characterize the onset of dystrophic phenotypes during myogenesis, and to separate the contribution of dystrophin’s signaling and structural roles to these deficits. This will identify the mechanism by which muscles become impaired during development. The second aim is to characterize the role dystrophin plays in the structure and function of the ASH neurons. These well-studied neurons will provide an amenable springboard to study the neuropathophysiology of Dmd. In the third aim, we will use our assay to identify downstream effectors of calmodulin responsible for the prevention of dystrophic phenotypes observed following reduction of calmodulin function in dys-1 animals. Identifying these effectors will be key to finding safe treatment avenues, sparing additional processes mediated by calmodulin. To validate our findings and bridge the gap to humans, we will use humanized dystrophic nematodes and human myogenic cell lines. Completion of these aims will provide key insights into Dmd pathophysiology and identify new molecular targets and pathways that can be used to treat this disease.

项目概要/摘要： 杜兴氏肌营养不良症（Dmd）是一种致死性退行性疾病，每5,000名男性中就有1人患病。DMD 是由编码肌营养不良蛋白的基因突变引起的，肌营养不良蛋白是一种高度保守的蛋白质， 膜与细胞外基质和其中的收缩机制。抗肌萎缩蛋白具有 结构和信号功能。肌营养不良蛋白的丢失与肌肉和神经退化有关。而 对小鼠、蠕虫和其他动物的传统分析通过功能丧失在遗传上模拟DMD 肌营养不良蛋白基因的突变，导致了巨大的进步，这些系统只产生相对 轻度肌肉和行为表型。迄今为止，还没有治愈DMD的方法。 为了在一个模型系统中模拟在DMD患者中观察到的急性肌肉变性， 我们开发了一种快速而廉价的线虫检测方法。我们的试验激发了强烈的行为和 营养不良（dys-1）线虫的细胞表型达到以前在其它系统中未达到的程度。 在上一个授予周期中，我们改进了检测方法，使其能够实现自动化， 候选治疗的通量筛选。我们继续描述了许多营养不良的表型 并发现它们首先出现在胚胎发育过程中。我们还发现了第一个神经损伤 在营养不良的蠕虫中，ASH神经元的感觉功能受损。抑制突变体，和 RNA干扰筛选都指向钙调蛋白作为治疗靶点。第一个具体目标是 该项目的目的是描述肌发生过程中营养不良表型的发病特征，并分离出 肌营养不良蛋白的信号传导和结构作用对这些缺陷的贡献。这将确定 肌肉在发育过程中受损的机制。第二个目的是描述 抗肌萎缩蛋白在ASH神经元的结构和功能中起作用。这些经过充分研究的神经元 为研究DMD的神经病理生理学提供了一个可靠的跳板。在第三个目标中，我们将使用 我们的测定鉴定了负责预防营养不良的钙调蛋白的下游效应物， 在dys-1动物中钙调蛋白功能降低后观察到的表型。识别这些 效应器将是寻找安全治疗途径的关键，避免由 钙调素为了验证我们的发现并弥合与人类之间的差距，我们将使用人源化营养不良的 线虫和人肌细胞系。这些目标的完成将为DMD提供关键的见解 病理生理学，并确定新的分子靶点和途径，可用于治疗这种疾病。

项目成果

Factors that influence magnetic orientation in Caenorhabditis elegans

• DOI: 10.1007/s00359-019-01364-y
• 发表时间: 2020-05-01
• 期刊: JOURNAL OF COMPARATIVE PHYSIOLOGY A-NEUROETHOLOGY SENSORY NEURAL AND BEHAVIORAL PHYSIOLOGY
• 影响因子: 2.1
• 作者: Bainbridge, C.;Clites, B. L.;Vidal-Gadea, A. G.
• 通讯作者: Vidal-Gadea, A. G.

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• DOI: 10.7554/elife.31414
• 发表时间: 2018
• 期刊: eLife
• 影响因子: 7.7
• 作者: Vidal-Gadea,Andres;Bainbridge,Chance;Clites,Ben;Palacios,BridgitteE;Bakhtiari,Layla;Gordon,Vernita;Pierce-Shimomura,Jonathan
• 通讯作者: Pierce-Shimomura,Jonathan