Screening for drug targets in a Drosophila model of muscle degeneration

在果蝇肌肉变性模型中筛选药物靶点

基本信息

批准号：
7800296
负责人：
Edward Owusu-Ansah
金额：
$ 5.22万
依托单位：
HARVARD MEDICAL SCHOOL
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-04-01 至 2011-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7800296
关键词：
Acquired Immunodeficiency Syndrome Address Apoptosis Atrophic Attenuated Autophagocytosis Biochemical Biological Assay Biological Markers Chromatin Clinical Code Data Degradation Pathway Diabetes Mellitus Drosophila genus Drug Delivery Systems Dystrophin Ecdysone Event Functional disorder Genes Genetic Genetic Screening Goals Heart failure Heat shock proteins Histones Human Human Genome Inherited Larva Link Luciferases Malignant Neoplasms Mediating Messenger RNA Modeling Molecular Molecular Profiling Molecular Target Muscle Muscle Weakness Muscular Atrophy Muscular Dystrophies Myopathy Organism Parkinsonian Disorders Pathway interactions Phosphoric Monoester Hydrolases Phosphotransferases Portraits Process Protein Isoforms Proteins Proteomics RNA Interference Relative (related person)Reporter Ribosomal Proteins Role Screening procedure Sepsis Signal Pathway Signal Transduction Sporadic Inclusion Body Myopathy Symptoms System Techniques Testing Transgenic Organisms Ubiquitin Uremia base ecdysone receptor fly genome-wide in vivo in vivo Model inhibitor/antagonist insulin signaling multicatalytic endopeptidase complex muscle degeneration muscle form parkin gene/protein protein expression public health relevance respiratory small molecule wasting

项目摘要

DESCRIPTION (provided by applicant): Muscular dystrophy refers to a diverse group of debilitating hereditary muscle disorders typified by a host of symptoms, including progressive muscular weakness and muscle wasting. To examine the molecular mechanisms that contribute to muscle loss, we have established a potent model of muscle degeneration in Drosophila. Constitutive expression of the ecdysone receptor (EcR) in somatic muscles of Drosophila larvae triggers their degeneration, thereby serving as a model for elucidating the molecular underpinnings of muscle atrophy/degeneration in a genetically tractable organism. Using this Drosophila system, we propose the following: Aim 1. Establish the relative contribution of catabolic and anabolic signals to EcR-mediated muscle wasting: Using a combination of genetic and biochemical techniques, we will examine the effect of inhibiting apoptosis, autophagy and degradation via the proteasomal pathway (i.e catabolic signals) as well as hyperstimulation of the insulin signaling network (anabolic signals) on EcR-mediated muscle degeneration; Aim 2. Identify universal biomarkers of muscle degeneration in flies and generate a "muscle degeneration reporter" for subsequent genetic screens: We will compare the mRNA and proteomic expression profiles of EcR-mediated muscle degeneration with that of three other well-established fly models of muscle degeneration (i.e. the dystrophin model, parkin/pinkl model and a sporadic inclusion-body myositis model) to identify several muscle pan-degeneration markers. Finally, we will develop a suitable assay for muscle degeneration, most likely a Luciferase transcriptional reporter based on one of the pan-degeneration markers; and Aim 3. Identify drug targets via a transgenic RNAi screen in muscles to identify suppressors of EcR- mediated muscle degeneration: Using publicly available RNAi transgenic lines, we will carry out a genome- wide in vivo RNAi screen (using the Luciferase-based reporter developed in Aim 2) in larval muscles to identify genes/pathways whose inhibition can ameliorate EcR-mediated muscle degeneration. PUBLIC HEALTH RELEVANCE: Given the high degree of conservation between the Drosophila and human genomes, we anticipate that molecular targets identified by this study will eventually provide leads for addressing muscle wasting/degeneration in humans. In summary, we seek to provide a genome-wide portrait of the molecular mechanisms that either trigger or suppress ecdysone-mediated muscle wasting in Drosophila, with the ultimate goal of identifying suitable drug targets for attenuating muscle degeneration in humans.

描述（由申请人提供）：肌肉营养不良是指一群具有多种症状的遗传性遗传性肌肉疾病，包括进行性肌肉无力和肌肉浪费。为了检查导致肌肉损失的分子机制，我们建立了果蝇中肌肉变性的有效模型。在果蝇幼虫的体细胞肌肉中，ecdysone受体（ECR）的组成型表达触发了它们的退化，从而是阐明遗传诱使生物中肌肉萎缩/退化的分子基础的模型。 Using this Drosophila system, we propose the following: Aim 1. Establish the relative contribution of catabolic and anabolic signals to EcR-mediated muscle wasting: Using a combination of genetic and biochemical techniques, we will examine the effect of inhibiting apoptosis, autophagy and degradation via the proteasomal pathway (i.e catabolic signals) as well as hyperstimulation of the insulin signaling network （合成代谢信号）ECR介导的肌肉变性；目的2。确定苍蝇中肌肉变性的普遍生物标志物，并产生一个“肌肉变性记者”，以进行后续的遗传筛查：我们将比较ECR介导的肌肉退化的mRNA和蛋白质组学表达谱与其他三种肌肉差异模型的肌肉模型和park蛋白模型A的肌肉模型，park蛋白模型，park蛋白模型，识别几个肌肉泛量化标记。最后，我们将开发一种适合肌肉变性的测定法，这很可能是基于泛量标记之一的荧光素酶转录报告基因。 and Aim 3. Identify drug targets via a transgenic RNAi screen in muscles to identify suppressors of EcR- mediated muscle degeneration: Using publicly available RNAi transgenic lines, we will carry out a genome- wide in vivo RNAi screen (using the Luciferase-based reporter developed in Aim 2) in larval muscles to identify genes/pathways whose inhibition can ameliorate EcR-mediated muscle degeneration.公共卫生相关性：鉴于果蝇和人类基因组之间的高度保护，我们预计这项研究确定的分子靶标最终将为解决人类的肌肉浪费/变性提供潜在客户。总而言之，我们试图提供全基因组的肖像，以触发或抑制果蝇中的ecdysone介导的肌肉浪费的分子机制，其最终目的是确定适合人类肌肉退化的药物靶标。