Whole body single AAV microgene therapy in canine DMD

犬 DMD 全身单一 AAV 微基因治疗

• 批准号: 9325083
• 负责人: Dongsheng Duan
• 金额: $ 60.03万
• 依托单位: UNIVERSITY OF MISSOURI-COLUMBIA
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-09-01 至 2020-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9325083
• 关键词: Accelerometer; Address; Adult; Affect; Animal Model; Attenuated; Autopsy; Becker Muscular Dystrophy; Binding; Biological Assay; Biology; Biopsy; Blood flow; Body Weight; Canis familiaris; Chronic; Clinical; Clinical Trials; Code; Complementary DNA; Data; Dependovirus; Disease; Dose; Duchenne muscular dystrophy; Dystrophin; Echocardiography; Electrocardiogram; Engineering; Extensor; Fibrosis; Forelimb; Foundations; Future; Gene Transduction Agent; Gene Transfer; Genes; Genome; Goals; Heart; Hindlimb; Histology; Human; Immune; Individual; Infiltration; Inflammation; Injection of therapeutic agent; Intramuscular Injections; Knowledge; Laboratories; Lead; Length; Life; Light; Limb structure; Magnetic Resonance Imaging; Mammals; Methods; Modeling; Mus; Muscle; Muscular Atrophy; Muscular Dystrophies; Mutation; Myopathy; Patients; Performance; Phenotype; Physiological; Preparation; Proteins; Protocols documentation; Quality of life; Regional Disease; Reporter Genes; Research Personnel; Safety; Skeletal Muscle; Systemic Therapy; T-Lymphocyte; Testing; Time; Toxic effect; Translations; Treatment Efficacy; Viral Vector; Wasting Syndrome; Wheelchairs; animal data; base; disease-causing mutation; dosage; gait examination; gene replacement therapy; gene therapy; immunogenic; immunoreaction; improved; intravenous injection; micro-dystrophin; mouse model; non-invasive monitor; novel; particle; prototype; public health relevance; response; systemic intervention; vector

 DESCRIPTION (provided by applicant) Duchenne muscular dystrophy (DMD) is caused by dystrophin deficiency. Gene replacement therapy holds great promise to treat DMD. Adeno-associated virus (AAV) is the leading viral vector for muscle gene therapy. However AAV has a small packaging capacity (~ 5-kb) and it cannot carry the full-length dystrophin cDNA (~12-kb). A microgene is a super-small synthetic dystrophin gene. It contains one-third of the full-length dystrophin coding sequence. Studies from many laboratories suggest that micro-dystrophin can effectively ameliorate muscle disease in dystrophin-deficient mice. Unfortunately, translation to large mammals has so far not been very successful. We recently engineered a new ∆R2-15/∆R18-19/∆R20-23/∆C microgene (abbreviated as ∆R2 µDys) and tested it adult dystrophin-null dogs by single muscle injection using Y731F AAV-9. Two months later, we observed dramatic reduction of inflammation and fibrosis. Importantly, eccentric contraction-induced damage, a physiological hallmark of DMD, was significantly alleviated. Our study demonstrates for the first time that microdystrophin can treat dystrophinopathy in muscles of large mammals. Our results also suggest that the newly developed Y731F AAV-9 ∆R2 µDys vector may hold great translational potential. The overarching goal of this proposal is to address key questions related to future clinical translatio. Specifically, we will pursue two aims including (1) to test the hypothesis that regional intramuscular injection can lead to persistent protection in adult dystrophic dogs. This set of studies will allow us to evaluate the feasibility of life-quality improving therapy in late-stage wheelchair-bound patients in the future; and (2) to test the hypothesis that a single intravenous injection can lead to bodywide amelioration in young dystrophic dogs. DMD affects all muscles in the body. This set of studies will allow us to determine whether systemic intervention can radically change the disease course in affected individuals. Taking together, our studies will break through major barriers in the field and provide the much-needed large animal data to guide human trials in the future.

 描述（由申请人提供） 杜氏肌营养不良 (DMD) 是由肌营养不良蛋白缺乏引起的，腺相关病毒 (AAV) 是肌肉基因治疗的主要病毒载体，但 AAV 的包装能力较小（~ 5-）。 kb），并且不能携带全长肌营养不良蛋白 cDNA（~12-kb）。微基因是一种超小的合成肌营养不良蛋白基因，它包含全长肌营养不良蛋白编码序列的三分之一。许多实验室的研究表明，微型肌营养不良蛋白可以有效改善肌营养不良蛋白缺陷小鼠的肌肉疾病，不幸的是，迄今为止，我们最近设计了一种新的 ΔR2-15/ΔR18-19/Δ 。 R20-23/ΔC 微基因（缩写为 ΔR2 µDys）并通过以下方法对成年抗肌营养不良蛋白缺失狗进行了测试两个月后，我们观察到使用 Y731F AAV-9 进行单次肌肉注射，炎症和纤维化显着减少，这是 DMD 的一个生理标志，我们的研究首次证明，微肌营养不良蛋白可以显着减轻。我们的结果还表明，新开发的 Y731F AAV-9 ΔR2 µDys 载体可能具有巨大的转化潜力。该提案的目的是解决与未来临床转化相关的关键问题，具体而言，我们将追求两个目标，包括（1）测试区域肌肉注射可以对成年营养不良犬产生持久保护的假设。评估未来晚期坐轮椅患者生活质量改善治疗的可行性；(2) 检验单次静脉注射可导致年轻营养不良犬全身改善的假设。身体。这一系列研究将使我们能够确定系统干预是否可以从根本上改变受影响个体的疾病进程。总之，我们的研究将突破该领域的主要障碍，并提供急需的大动物数据来指导人体试验。未来。