Antisense oligonucleotide suppression of non-deletion DMD causing mutations

反义寡核苷酸抑制非缺失 DMD 引起的突变

• 批准号: 8044694
• 负责人: STEPHEN D WILTON
• 金额: $ 22.83万
• 依托单位: UNIVERSITY OF WESTERN AUSTRALIA
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-01-01 至 2013-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8044694
• 关键词: Actins; Address; Affect; Animal Model; Antisense Oligonucleotides; Architecture; Becker Muscular Dystrophy; Binding; Binding Sites; Cell Line; Cells; Computer Simulation; Custom; DNA; Defect; Deletion Mutation; Development; Disease; Distant; Duchenne muscular dystrophy; Dystroglycan; Dystrophin; Excision; Exhibits; Exons; Gene Expression; Gene Expression Profile; Genes; Genetic; Genetic Polymorphism; Genomics; Glycoproteins; Human; In Vitro; Individual; Lead; Lesion; Life Expectancy; Link; Messenger RNA; Morbidity - disease rate; Morphology; Mus; Muscle; Mutate; Mutation; Neonatal; Nonsense Mutation; Patients; Phenotype; Process; Protein Isoforms; Proteins; RNA Splicing; Reading Frames; Respiratory Diaphragm; Severities; Site; Structure; Testing; Transcript; Translations; Variant; base; design; effective therapy; insertion/deletion mutation; mRNA Precursor; muscle strength; premature; public health relevance; restoration; wasting

DESCRIPTION (provided by applicant): Duchenne muscular dystrophy (DMD) is a fatal X-linked muscle-wasting disorder caused by protein truncating mutations in the dystrophin gene. Antisense oligomer induced removal of an exon carrying a nonsense mutation, or exons flanking frame-shifting deletions, the most common type of DMD mutation, has been shown to generate an in-frame message and an internally deleted, but functional protein. Becker muscular dystrophy (BMD) is an allelic disorder typically caused by in-frame deletions of one or more exons, most commonly in the first two thirds of the gene. The severity of BMD varies from borderline DMD to asymptomatic, and the dystrophin genes in mildly affected BMD patients provide an indication of functional exon combinations. At least one third of DMD cases result from duplications, micro-insertions/deletions and single base changes that alter splice site recognition or cause premature termination of translation. This project will address the design and application of antisense oligomers for induced exon skipping, for those DMD cases caused by non-deletion mutations. Patient cell lines will be transfected with test compounds and exon skipping assessed. Exon skipping strategies will be modified to maximize induced dystrophin quality and quantity, as permitted by the context of each particular dystrophin gene lesion. The specific aims are to: 7 Optimise antisense oligomers to remove exons carrying sequence variations (disease-causing or neutral polymorphims) that would otherwise compromise exon skipping. 7 Develop exon skipping strategies appropriate to DMD cases caused by pseudo-exon incorporation or duplications of one or more exons. 7 Develop transient animal models to identify functionally significant dystrophin domains, according to exon boundaries, to facilitate design of optimal exon skipping strategies. PUBLIC HEALTH RELEVANCE: Duchenne muscular dystrophy is a relentlessly progressive, fatal disease for which there is no effective treatment. Specific exon removal has the potential to greatly reduce the severity of DMD, and restoration of dystrophin expression, even of partial function in a DMD patient is expected to result in a BMD-like phenotype, and reduce morbidity and extend life expectancy. This application seeks to develop personalised exon skipping therapies for the one third of DMD patients who have non-deletion mutations. Exon skipping should be made available to all patients who could benefit, not only those with the more common exon deletion mutations.

描述（由申请人提供）： 杜氏肌营养不良症 (DMD) 是一种致命的 X 连锁肌肉萎缩性疾病，由肌营养不良蛋白基因的蛋白质截短突变引起。反义寡聚体诱导去除携带无义突变的外显子，或移码侧翼缺失（最常见的 DMD 突变类型）的外显子，已被证明可生成框内消息和内部删除但有功能的蛋白质。贝克尔肌营养不良症 (BMD) 是一种等位基因疾病，通常由一个或多个外显子的框内缺失引起，最常见的是基因的前三分之二。 BMD 的严重程度从临界 DMD 到无症状不等，轻度受影响的 BMD 患者的肌营养不良蛋白基因提供了功能性外显子组合的指示。至少三分之一的 DMD 病例是由重复、微插入/缺失和单碱基变化引起的，这些变化改变了剪接位点识别或导致翻译过早终止。该项目将针对非缺失突变引起的 DMD 病例，解决诱导外显子跳跃的反义寡聚物的设计和应用。将用测试化合物转染患者细胞系并评估外显子跳跃。在每个特定肌营养不良蛋白基因损伤的背景允许的情况下，将修改外显子跳跃策略，以最大限度地提高诱导肌营养不良蛋白的质量和数量。具体目标是： 7 优化反义寡聚物以去除携带序列变异（致病或多态性）的外显子，否则会影响外显子跳跃。 7 制定适合由假外显子掺入或一个或多个外显子重复引起的 DMD 病例的外显子跳跃策略。 7 开发瞬时动物模型，根据外显子边界识别功能上重要的肌营养不良蛋白结构域，以促进最佳外显子跳跃策略的设计。 公共卫生相关性： 杜氏肌营养不良症是一种持续进展的致命疾病，目前尚无有效的治疗方法。特定的外显子去除有可能大大降低 DMD 的严重程度，并且恢复肌营养不良蛋白的表达，甚至恢复 DMD 患者的部分功能，预计将导致 BMD 样表型，并降低发病率并延长预期寿命。该申请旨在为三分之一具有非缺失突变的 DMD 患者开发个性化的外显子跳跃疗法。外显子跳跃应该提供给所有可以受益的患者，而不仅仅是那些具有更常见的外显子缺失突变的患者。