Whole body single AAV microgene therapy in canine DMD

犬 DMD 全身单一 AAV 微基因治疗

基本信息

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
项目类别：
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

项目摘要

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） Duchenne肌肉营养不良（DMD）是由肌营养不良蛋白缺乏引起的。基因替代疗法有很好的希望治疗DMD。腺相关病毒（AAV）是肌肉基因治疗的主要病毒载体。但是，AAV具有较小的包装能力（〜5-kb），并且无法携带全长肌营养不良蛋白cDNA（〜12-kb）。小币是一种超级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。来自许多实验室的研究表明，微型障碍蛋白可以有效地改善肌营养不良蛋白缺陷型小鼠的肌肉疾病。不幸的是，到目前为止，转化为大型哺乳动物还不是很成功。我们最近设计了新的∆R2-15/∆R18-19/∆R20-23/∆C微胶（缩写为∆R2 µDys），并使用Y731F AAV-9通过单肌注射单肌肉注射来测试其成年肌营养不良的狗。两个月后，我们观察到感染和纤维化的显着减少。重要的是，偏心收缩引起的损害是DMD的物理标志，可显着缓解。我们的研究首次表明，微隔膜蛋白可以治疗大型哺乳动物肌肉中的肌营养不良。我们的结果还表明，新开发的Y731F AAV-9 ∆R2 µDys向量可能具有巨大的转化潜力。该提案的总体目标是解决与未来临床翻译有关的关键问题。具体而言，我们将追求两个目标，包括（1）测试肌肉内注射可以导致成人营养不良犬持续保护的假设。这套研究将使我们能够评估后期坐轮椅结合的患者的生活质量改善治疗的可行性；（2）测试单个静脉注射可以导致年轻营养不良犬的整体改善的假设。 DMD会影响体内的所有肌肉。这组研究将使我们能够确定系统性干预是否可以从根本上改变受影响的个体的疾病病程。综合起来，我们的研究将打破该领域的主要障碍，并提供急需的大型动物数据，以指导人类的试验。