Dystrophin turnover after exon-skipping

外显子跳跃后肌营养不良蛋白的周转

基本信息

批准号：
9293874
负责人：
KANNEBOYINA NAGARAJU
金额：
$ 39万
依托单位：
STATE UNIVERSITY OF NY,BINGHAMTON
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-09-01 至 2019-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9293874
关键词：
Address Affect Antisense Oligonucleotides Attention Becker Muscular Dystrophy Bromodeoxyuridine Cells Chemistry Clinical Trials Confounding Factors (Epidemiology)Data Development Disease Duchenne muscular dystrophy Dystrophin Dystrophin-Associated Proteins Elements Environment Exhibits Exons Female Fiber Genes Genetic Genetic Transcription Genomics Human Individual Inherited Investigation Label Laboratories Length Link Longevity Maintenance Mass Spectrum Analysis Measures Methods Mus Muscle Muscle Fibers Muscle Proteins Mutation Myocardium Myopathy Oligonucleotides Open Reading Frames Pathology Patients Physiologic pulse Process Production Protein Analysis Proteins Proteomics Protocols documentation Reagent Sampling Severities Severity of illness Skeletal Muscle System Techniques Therapeutic Transcript Translating Translations Treatment Protocols Variant Vertebral column abstracting base boys design exon skipping improved in vivo knowledge base mdx mouse mouse model mutant protein function research study response restoration stable isotope success tool

项目摘要

Abstract Duchenne muscular dystrophy (DMD) is the commonest lethal inherited X-linked myopathy. The most promising approach to treating this condition, is by use of antisense agents that promote skipping of exons that disrupt open reading frame so as to provide a translatable form of the transcript, thus restoring expression of the missing dystrophin protein. Current clinical trials of this idea have given results that are compromised by low yields of dystrophin and inter-sample variation. This proposal aims to evaluate the fundamental elements of the processes that link exon-skipping to the production and maintenance of functional amounts of dystrophin in muscles. To do this we will investigate the effects of exon-skipping within the mutant dystrophin gene on the production of the 3 different truncated dystrophin proteins produced by skipping different exons and measure the stability and longevity of these proteins within skeletal and cardiac muscles of the dystrophic mdx mouse. We will use the mdx mouse in which exon 52 has been deleted from the dystrophin gene because this mutation presents a number of options for restoring open reading frame and dystrophin production by skipping of different exons that are the main targets of human trials. By use of Mass Spectroscopy techniques recently developed within our laboratory we can accurately measuring the amounts of dystrophin and dystrophin-associated proteins, and will follow the dynamics of their production, stability and turnover. Our aim is to relate these measures to the extent of alleviation of pathology associated with the restoration of 3 different these proteins. This will be the first investigation of the dynamics of this class of muscle proteins in muscles in vivo and will provide methods and protocols that will be more widely applicable. On the basis of this knowledge it should be possible to devise optimal protocols for application of exon-skipping to DMD boys in clinical trials. It will also provide an objective method of selecting the best exons and combinations of exons to skip as the progressively improving antisense reagents become available.

抽象的杜氏肌营养不良症 (DMD) 是最常见的致命性遗传性 X 连锁遗传病肌病。治疗这种情况最有希望的方法是使用反义促进破坏开放阅读框的外显子跳跃的试剂，从而提供转录物的可翻译形式，从而恢复缺失的肌营养不良蛋白的表达蛋白质。目前这一想法的临床试验给出的结果受到低浓度的影响抗肌萎缩蛋白的产量和样本间变异。该提案旨在评估将外显子跳跃与生产联系起来的过程的基本要素维持肌肉中抗肌营养不良蛋白的功能量。为此，我们将研究突变肌营养不良蛋白基因内外显子跳跃对通过跳过不同的步骤产生 3 种不同的截短肌营养不良蛋白外显子并测量这些蛋白质在骨骼和骨骼中的稳定性和寿命营养不良的 mdx 小鼠的心肌。我们将使用 mdx 鼠标，其中外显子 52 已从肌营养不良蛋白基因中删除，因为该突变呈现出用于恢复开放阅读框和肌营养不良蛋白产生的选项数量跳过作为人体试验主要目标的不同外显子。通过使用质量我们实验室最近开发的光谱技术我们可以准确地测量肌营养不良蛋白和肌营养不良蛋白相关蛋白的量，并将遵循他们的生产动态、稳定性和营业额。我们的目标是将这些联系起来采取措施减轻与恢复相关的病理状况 3 这些蛋白质不同。这将是对此类动态的首次调查体内肌肉中的肌肉蛋白，并将提供将被适用范围更广。在这些知识的基础上，应该可以设计出在临床试验中对 DMD 男孩应用外显子跳跃的最佳方案。它将还提供了选择最佳外显子和外显子组合的客观方法随着反义试剂的不断改进，可以跳过。