Therapeutic target discovery in Drosophila models of Nemaline Myopathy

线形肌病果蝇模型中治疗靶点的发现

• 批准号: 9001905
• 负责人: MARY K BAYLIES
• 金额: $ 23.22万
• 依托单位: SLOAN-KETTERING INST CAN RESEARCH
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-03-01 至 2017-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9001905
• 关键词: ACTA1 gene; Address; Adult; Affect; Biological Assay; Cessation of life; Clinic; Collaborations; Data; Development; Diagnosis; Disease; Disease Progression; Disease model; Drosophila genus; Drug Combinations; Drug Targeting; Effectiveness; FDA approved; Future; Gene Expression; Gene Expression Profiling; Gene Mutation; Generations; Genes; Genetic; Goals; Health; Heterogeneity; Incidence; Libraries; Life; Light; Link; Location; Methods; Modeling; Molecular Profiling; Mus; Muscle; Muscle Weakness; Muscle function; Mutation; Myopathy; Nemaline Myopathies; Neonatal; Onset of illness; Orthologous Gene; Patients; Penetrance; Pharmaceutical Preparations; Phenotype; Play; Population; Proteins; Reproducibility; Research; Role; Sarcomeres; Severities; Severity of illness; Skeletal Muscle; Specific qualifier value; Sum; Symptoms; Testing; Therapeutic; Thin Filament; Time; Validation; Work; base; clinically relevant; cofilin 2; congenital myopathy; cost effective; disease heterogeneity; drug development; effective therapy; fly; genetic analysis; genetic manipulation; genetic signature; high throughput screening; improved; in vivo; insight; knock-down; muscle regeneration; new therapeutic target; novel; premature; screening; therapeutic target; tool

DESCRIPTION (provided by applicant): Our long-term goal is to find effective therapeutics for treatment of Nemaline myopathy (NM). NM is the most common form of non-dystrophic congenital myopathy, having an incidence of 1/20,000 in some populations. It is a slow or non-progressive disease, characterized by the presence of Nemaline bodies in the affected muscle tissue, by muscle weakness and by the absence of muscle regeneration. NM is a heterogeneous group of myopathies of variable severity: different types of NM can range from neonatal lethal to mild adult onset. To date, there is no effective treatment for this disease. Mutations in nine different genes have been linked to NM, with six of these encoding skeletal muscle sarcomere thin filament-associated proteins, including Cofilin 2 (CFL2). No relationships exist between gene mutations and disease severity or other phenotypes. Gene expression profiling of a heterogeneous group of NM patients uncovered a consistent signature for NM; how these changes in gene expression contribute to disease onset and/or severity is not clear. We have developed a novel Drosophila model of NM, using mutations in the Drosophila CFL2 ortholog, twinstar (tsr). This model displays the hallmarks of NM: Nemaline bodies, muscle weakness and, in the severe condition, premature death. In our less severe condition, muscle function is reduced without gross alterations to the sarcomere. We will use our novel model to address the critical gaps in both available treatments for this disease and in our understanding of the contributions of NM signature genes to disease development and severity. Taking advantage of the fast generation time, reproducibility and penetrance of the phenotype, the genetic tools available, and the ability to perform cost effective, relatively high throughput screens, we propose two aims: In the first, we will screen an FDA approved drug library to identify drugs that delay or eliminate NM symptoms. These studies depend on our robust model, our recently developed methods for drug application, and our assays streamlined for such a high throughput screen. We expect to find FDA approved drugs that impact the formation or severity of NM. In the second, we will determine how NM signature genes contribute to NM disease severity. We will first assess gene expression profiles in our mild and strong NM conditions. After validation, we will compare this set of genes with patient NM data generated by our collaborator, Dr. Alan Beggs. Conserved genes that are similarly regulated will then be analyzed for their contribution to disease in our fly model. These data will enhance our understanding of NM heterogeneity, provide insights to drug effectiveness and supply targets for future drug development. Together the work outlined in this proposal with shed new light on NM: we will identify a short list of drugs and clinically relevant genes that can be validated, via our collaboration with Dr. Beggs, first in vertebrate and mammalian disease models and, ultimately, in patients.

描述（由申请人提供）：我们的长期目标是找到治疗线状体肌病（NM）的有效疗法。NM是非营养不良性先天性肌病的最常见形式，在某些人群中的发病率为1/20，000。它是一种缓慢或非进行性疾病，其特征在于受影响的肌肉组织中存在线状体，肌肉无力和缺乏肌肉再生。NM是一组不同严重程度的肌病：不同类型的NM范围从新生儿致死到轻度成人发作。迄今为止，对这种疾病没有有效的治疗方法。九种不同基因的突变与NM有关，其中六种编码骨骼肌肌小节细丝相关蛋白，包括Cofilin 2（CFL 2）。基因突变与疾病严重程度或其他表型之间不存在关系。一组异质性NM患者的基因表达谱揭示了NM的一致特征;这些基因表达变化如何导致疾病发作和/或严重程度尚不清楚。我们已经开发了一种新的果蝇模型NM，在果蝇CFL 2直系同源物，双星（TSR）的突变。该模型显示了NM的特征：线状体，肌肉无力，在严重的情况下，过早死亡。在我们不太严重的情况下，肌肉功能降低，而肌节没有明显改变。我们将使用我们的新模型来解决这种疾病的现有治疗方法以及我们对NM特征基因对疾病发展和严重程度的贡献的理解中的关键差距。利用表型的快速生成时间、再现性和重复性、可用的遗传工具以及进行成本有效的、相对高通量筛选的能力，我们提出两个目标：首先，我们将筛选FDA批准的药物库以鉴定延迟或消除NM症状的药物。这些研究依赖于我们强大的模型，我们最近开发的药物应用方法，以及我们为这种高通量筛选而简化的分析。我们希望找到FDA批准的影响NM形成或严重程度的药物。在第二，我们将确定如何NM签名基因有助于NM疾病的严重程度。我们将首先评估我们的温和和强烈的NM条件下的基因表达谱。验证后，我们将比较这组基因与我们的合作者Alan Beggs博士生成的患者NM数据。然后将在我们的苍蝇模型中分析受到类似调控的保守基因对疾病的贡献。这些数据将增强我们对NM异质性的理解，为药物有效性提供见解，并为未来的药物开发提供目标。这项提案中概述的工作与NM的新进展一起：我们将通过我们的研究，确定一个可以验证的药物和临床相关基因的短名单。 与Beggs博士合作，首先在脊椎动物和哺乳动物疾病模型中，最终在患者中。