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Safety and Efficacy of Systemic Gene Therapy in Informative Models for DMD

DMD 信息模型中系统基因治疗的安全性和有效性

基本信息

批准号：
9149074
负责人：
HANSELL H STEDMAN
金额：
$ 57.15万
依托单位：
UNIVERSITY OF PENNSYLVANIA
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：
2015-09-30 至 2020-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9149074
关键词：
Accounting Activities of Daily Living Address Affect Age of Onset Antigens Appearance Attention Benefits and Risks Biological Assay Blinded Body Weight Canis familiaris Capsid Cardiac Clinical Clinical Trials Cytotoxic T-Lymphocytes Dependence Dependovirus Disease Disease model Dog Diseases Dose Dose-Limiting Duchenne muscular dystrophy Dystrophin Dystrophin-Associated Protein Complex Educational workshop Event Exercise Family Caregiver Ferritin Fiber Forelimb Future Gene Deletion Gene Transfer Genes German population Growth Heart High Prevalence Hindlimb Human Immune response Immunosuppression Incidence Individual Inherited Invaded Measurement Measures Mendelian disorder Methods Modeling Motor Activity Movement Mus Muscle Muscle Fibers Muscle function Muscular Dystrophies Mutation Myositis National Institute of Neurological Disorders and Stroke Neonatal Newborn Animals Organ Peptides Peripheral Phase Phase I Clinical Trials Phenotype Physically Handicapped Principal Investigator Protein Deficiency Proteins Pseudogenes Reaction Recombinants Recovery Regulation Research Subjects Resources Respiratory Diaphragm Respiratory physiology Risk Running Safety Sequence Deletion Signs and Symptoms T-Lymphocyte Technology Testing Therapeutic Thymus Gland Time Tissues Toxic effect Transgenes Treatment Efficacy Utrophin adeno-associated viral vector base boys burden of illness cell mediated immune response central tolerance clinically relevant design fetal gene therapy gene therapy clinical trial grasp homologous recombination immunotoxicity in vivo indexing innovation mdx mouse micro-dystrophin miniaturize mortality muscle form muscle strength nonhuman primate novel peripheral tolerance polypeptide pre-clinical preclinical study prevent programs public health relevance respiratory response screening skeletal vector

项目摘要

DESCRIPTION (provided by applicant): Duchenne Muscular Dystrophy (DMD) is caused by inherited deficiency of the protein dystrophin. The overall aim of this project is to build towards rational clinical trials of gene therapy in DMD, following recent studies showing unprecedented protection of muscle fibers by a single dose of a novel vector in dystrophic mice and dogs. To minimize the risk of an immunological reaction against the transgene product, we have substituted utrophin for dystrophin. A systematically optimized, synthetic utrophin minigene is used with a muscle-targeting vector, AAV9, to achieve systemic gene transfer in newborn animals. In mdx mice following subsequent growth to maturity, the histological appearance of the treated muscle is restored to normal; in Aim 1 we now test whether this level of muscle fiber protection will correct muscle function in a wide range of clinically relevant assays. We use appropriate methods to study locomotor activity of the mice, types of movement, voluntary running times and distances, as well as forelimb and hindlimb muscle strength in grip tests. We assay for normalization of the CPK following exercise. Finally, we establish the function of the recombinant utrophin in isolated muscles, including the diaphragm and the heart, post mortem. To address the major safety issue facing potential subjects in clinical trials, in Aim 2 we turn ou attention to the immune response, using a unique disease model afforded by a naturally occurring deletion in the dog. The index mutation was identified in a German Short Haired Pointer (GSHPMD), and was shown by our lab to result from a homologous recombination event between ferritin-like pseudogenes flanking the entire canine dystrophin gene. We systematically investigate the immunological response to recombinant dystrophin and utrophin in this model to rigorously test the hypothesis that in the absence of central (thymic) immunological tolerance, host T cells will invade AAV-transduced muscle fibers, thereby eliminating recombinant dystrophin expression and causing a clinically severe myositis that exacerbates the clinical course of GSHPMD. We further hypothesize that central immunological tolerance will protect muscle fibers transduced with recombinant utrophin, thereby facilitating therapeutic efficacy. Finally, in Aim 3 we conduct a dose-finding study of systemic vector delivery in the dystrophic dog to estimate the appropriate dose for later use in a preclinical tria of efficacy. This will require the use of unprecedented titers of AAV in individual dogs, based on our studies thus far in mice, and will include screening tests for vector toxicity.

描述(申请人提供)：Duchenne肌营养不良症(DMD)是由Duchenne肌营养不良蛋白遗传缺陷引起的。该项目的总体目标是为实现 DMD基因治疗的合理临床试验，此前最近的研究表明，单剂新型载体对营养不良的小鼠和狗的肌肉纤维具有前所未有的保护作用。为了最大限度地减少针对转基因产物的免疫反应的风险，我们用优营养素代替了营养不良蛋白。一种系统优化的人工合成的微促性腺激素基因与肌肉靶向载体AAV9一起用于在新生动物中实现系统的基因转移。在随后生长到成熟的MDX小鼠中，经过处理的肌肉的组织外观恢复到正常；在目标1中，我们现在测试这种水平的肌肉纤维保护是否将在广泛的临床相关分析中纠正肌肉功能。我们使用适当的方法来研究小鼠的运动活动、运动类型、随意跑的时间和距离，以及握力测试中的前肢和后肢肌肉力量。我们测定运动后CPK的正常化程度。最后，我们在死后的分离肌肉中，包括横隔肌和心脏，建立了重组人促性腺激素的功能。为了解决临床试验中潜在受试者面临的主要安全问题，在目标2中，我们将注意力转向免疫反应，使用一种独特的疾病模型，该模型由狗体内自然发生的缺失提供。该索引突变是在一只德国短毛指针(GSHPMD)中发现的，我们的实验室证明该突变是由于犬营养不良蛋白基因两侧的铁蛋白样假基因之间的同源重组事件造成的。我们在这个模型中系统地研究了重组抗肌营养不良蛋白和中性粒细胞营养不良蛋白的免疫学反应，以严格检验这一假设，即在缺乏中枢(胸腺)免疫耐受的情况下，宿主T细胞将入侵AAV转导的肌肉纤维，从而消除重组抗肌营养不良蛋白的表达，并导致临床上严重的肌炎，从而加剧GSHPMD的临床病程。我们进一步假设中枢免疫耐受将保护重组人促性腺激素转导的肌肉纤维，从而促进治疗效果。最后，在目标3中，我们在营养不良的狗身上进行了一项全身媒介传递的剂量发现研究，以估计在临床前疗效试验中稍后使用的适当剂量。根据我们迄今在小鼠身上的研究，这将需要在单个狗身上使用前所未有的AAV滴度，并将包括对媒介毒性的筛选测试。