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（由应用提供）：Duchenne肌肉营养不良（DMD）是由蛋白质肌营养不良蛋白的遗传缺乏引起的。该项目的总体目的是建立最近的研究表明，DMD中基因治疗的理性临床试验表明，单剂量的营养不良小鼠和狗中的单剂量对肌肉纤维进行了前所未有的保护。为了最大程度地减少对转化产物的免疫反应的风险，我们用伊特罗蛋白代替了肌营养不良蛋白。系统优化的合成伊特罗芬微基因与肌肉靶向载体AAV9一起使用，以实现新生动物的全身基因转移。 MDX小鼠随后生长到成熟度后，治疗的肌肉的组织学外观恢复到正常状态。在AIM 1中，我们现在测试这种肌肉纤维保护水平是否会在广泛的临床相关测定中纠正肌肉功能。我们使用适当的方法来研究小鼠的运动活动，运动类型，自愿跑步时间和距离，以及握把测试中的前肢和后肢肌肉力量。我们分析运动后CPK的标准化。最后，我们在孤立的肌肉中建立了重组乌托蛋白的功能，包括diaphragm and the the the the the Mortem。为了解决临床试验中潜在受试者面临的主要安全问题，在AIM 2中，我们使用狗的自然缺失提供了独特的疾病模型，将其关注对免疫反应。指数突变是在德国短头发指针（GSHPMD）中鉴定出来的，我们的实验室表明，是由于铁蛋白样的假基因之间的同源重组事件而导致的。在该模型中，我们系统地研究了对重组肌营养不良蛋白和紫蛋白的免疫学反应，以严格检验以下假设：在缺乏中心（胸腺）免疫耐受性的情况下，宿主T细胞将入侵AAV转移的肌肉纤维，从而消除重组的临床上的临时性疾病，从而消除临床上的临床症状，并导致临床上的临床症状。我们进一步假设中央免疫耐受性将保护用重组乌托蛋白转移的肌肉纤维，从而支持治疗效率。最后，在AIM 3中，我们对营养不良狗的全身载体传递进行了剂量调查研究，以估算适当的剂量，以便以后在临床前效率三亚临床三学三脱。这将需要根据迄今为止在小鼠中的研究中使用AAV前所未有的滴度，并包括对媒介毒性的筛查测试。