Pathogenesis and Treatment of Muscular Dystrophy

肌营养不良症的发病机制和治疗

• 批准号: 7488889
• 负责人: LOUIS M KUNKEL
• 金额: $ 147.05万
• 依托单位: BOSTON CHILDREN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-09-25 至 2012-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7488889
• 关键词: Pathogenesis; Treatment; Muscular; Dystrophy

Description (provided by applicant): Over the last 5 years, we have been working to establish the zebrafish as a model for muscular dystrophy. In this capacity, we have published the phenotype of zebrafish lacking dystrophin and 5- sarcoglycan, completed a large genetic screen to isolate additional dystrophic mutants, and identified the mutant gene in the runzel mutant. Using our experiences in muscle research and in establishing the zebrafish as a disease model, we now propose to use the fish to investigate the pathogenesis of muscular dystrophy and evaluate cell therapy as a potential treatment option. The first aim in this project proposes to fully characterize one of our available dystrophic zebrafish models with the goal of better understanding the pathogenesis of muscular dystrophy. We have selected the emz mutant for further analysis since its mutation rough maps to a genomic interval void of any genes orthologous to those currently associated with muscular dystrophy. This mutant shows a phenotype very similar to the dystrophin mutant (sapje) suggesting that the emz phenotype of muscle degeneration is symptomatic of muscular dystrophy. We propose to identify the genetic mutation in this mutant using a traditional mapping approach and then sequencing candidate genes to identify the specific mutation. If the orthologous human gene is not currently associated with muscular dystrophy, the gene will be considered a disease candidate and sequenced in human patients for which the cause of muscular dystrophy is unknown. Since mutations in seemingly unrelated proteins can manifest as muscular dystrophy, the identification of additional genes would be helpful for establishing disease pathways. Secondly, we have established, methods to transplant cell populations in zebrafish at all developmental stages and now propose using this system to identify the cell population most capable of engrafting into and correcting the diseased muscle. Gene expression profiles of muscle engrafting cell populations will be compared with non-engrafting cells to identify genes expressed predominantly in the engrafting cells. Differentially expressed genes will be considered potential markers and used to purify analogous cell populations in mammals for future experimentation and therapy. Finally, we plan to dissect the lineage relationship of various stem cell populations by assaying the developmental potential of zebrafish muscle progenitor cells. This will be accomplished by transplanting limited populations of labeled cells early in development and then following their fate as the fish matures.

描述（由申请人提供）：在过去的5年中，我们一直在努力建立斑马鱼作为肌肉营养不良的模型。以这种能力，我们发表了缺乏肌营养不良蛋白和5-萨科利果的斑马鱼的表型，完成了一个大遗传筛选，以分离额外的营养不良突变体，并鉴定出Runzel突变体中的突变基因。利用我们在肌肉研究中的经验并确立斑马鱼作为疾病模型，我们现在建议使用鱼研究肌肉营养不良的发病机理，并将细胞疗法评估为潜在的治疗选择。 该项目的第一个目的是为了更好地理解肌肉营养不良的发病机理的目的是充分表征我们可用的营养不良斑马鱼模型之一。我们已经选择了EMZ突变体进行进一步分析，因为其突变图粗图与目前与肌肉营养不良相关的基因的基因组间隔无效。该突变体显示出与肌营养不良蛋白突变体（SAPJE）非常相似的表型，这表明肌肉退化的EMZ表型是肌肉营养不良的症状。我们建议使用传统的映射方法鉴定该突变体中的遗传突变，然后对候选基因进行测序以鉴定特定的突变。如果直系同源的人类基因目前与肌营养不良症无关，则该基因将被视为疾病候选者，并在人类患者中被测序，肌营养不良的原因尚不清楚。由于看似无关的蛋白质突变可以表现为肌肉营养不良，因此鉴定其他基因将有助于建立疾病途径。 其次，我们已经建立了在所有发育阶段移植细胞种群的方法，现在建议使用该系统来识别最有能力灌输和纠正患病肌肉的细胞群。将肌肉雕刻细胞群体的基因表达谱与非碎片细胞进行比较，以鉴定在雕刻细胞中主要表达的基因。差异表达的基因将被视为潜在标记，并用于纯化哺乳动物中的类似细胞群，以进行未来的实验和治疗。最后，我们计划通过分析斑马鱼肌肉祖细胞的发育潜力来剖析各种干细胞种群的谱系关系。这将通过在发育的早期移植有限的标记细胞种群，然后随着鱼的成熟而命运来实现。