Optimizing and validation of gene therapy vectors to treat limb girdle muscular dystophy

治疗肢带型肌营养不良症的基因治疗载体的优化和验证

• 批准号: 10219370
• 负责人: JEFFREY S CHAMBERLAIN
• 金额: $ 55.28万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-08-01 至 2025-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10219370
• 关键词: Affect; Animal Model; Antibodies; Area; Binding Sites; Calpain; Cardiac; Cardiotoxicity; Clinical Trials; Code; Codon Nucleotides; Collaborations; Complementary DNA; Development; Diagnosis; Disease; Dose; Duchenne muscular dystrophy; Dystrophin; Exercise Tolerance; Fiber; Gene Delivery; Gene Expression; Gene Transduction Agent; Gene therapy trial; Genes; Goals; Heart; Heart Block; Human; International; Knockout Mice; Knowledge; Limb structure; Limb-Girdle Muscular Dystrophies; Longitudinal Studies; MicroRNAs; Modification; Mus; Muscle; Muscle Weakness; Muscular Atrophy; Muscular Dystrophies; Mutation; Myocardium; Neuromuscular Diseases; Outcome; Outcome Measure; Pathogenesis; Patients; Peptide Hydrolases; Phenotype; Physiological; Property; Proteins; Series; Skeletal Muscle; Testing; Therapeutic; Therapeutic Effect; Therapeutic Intervention; Time; Toxic effect; Transgenes; Treatment Efficacy; Universities; Untranslated Regions; Validation; Variant; Viral Vector; Washington; Wasting Syndrome; Western Blotting; Wheelchairs; Work; adeno-associated viral vector; design; differential expression; experience; gene therapy; gene therapy clinical trial; in vitro testing; in vivo; laser capture microdissection; member; micro-dystrophin; mini-dystrophin; mitochondrial dysfunction; mouse model; novel; overexpression; pre-clinical; safety testing; therapeutic evaluation; vector

PROJECT SUMMARY We aim to develop a gene therapy for limb girdle muscular dystrophy type 2A (LGMD2A), an autosomal recessive (AR) muscle wasting disorder due to mutations in CAPN3. LGMD2A is considered to be the most prevalent of the AR LGMDs and yet there is currently no treatment for patients, who are usually wheelchair dependent a decade after diagnosis. We and others have shown that overexpression of CAPN3 can be accomplished in skeletal muscle without toxicity; a finding which makes the feasibility of gene therapy for LGMD2A a realistic goal; however, LGMD2A is unique from most other LGMDs and this fact warrants careful development of gene therapy vectors. One consideration is that CAPN3 cannot be expressed in the heart, due to cardiac toxicity, which is not the case with Duchenne muscular dystrophy and other LGMDs. Although a few pre-clinical, proof of concept studies have successfully accomplished AAV-Capn3 overexpression in mice, there has not been a systematic optimization of any gene therapy construct for humans with LGMD2A, especially one that considers the relative skeletal muscle vs cardiac gene expression issues. Furthermore, because LGMD2A preferentially impacts slow fibers, it is critical that the therapeutic construct that is ultimately carried into clinical trials is optimized for slow fiber expression. Because each patient can only be dosed one time, it is imperative that the construct used for gene delivery is ideal. In this application, we will use an iterative and systematic approach to optimize these vectors. We have assembled an expert, collaborative team with deep experience in the area of regulatory cassettes, AAV vector development and LGMD2A pathogenesis and calpainopathy animal models. The team will work together to generate and test the safety and efficacy of a series of AAV vectors to develop a treatment for LGMD2A. These vectors will be optimized for slow fiber expression while avoiding cardiac toxicity. One team member, Dr. Hauschka, is largely credited with creating the vast majority of regulatory cassettes being used in the current AAV-gene therapy trials for Duchenne muscular dystrophy. Dr. Chamberlain designed and optimized the first micro and mini dystrophins, which formed the basis for all constructs currently in gene therapy clinical trials for DMD. Drs. Spencer and Kramerova generated numerous mouse models that have led to the identification of outcome measures useful for testing therapeutic interventions for LGMD2A. Dr. Cannon, is an internationally known muscle physiologist who will carry out physiological assessments of contractile function. The team will apply their extensive and cumulative knowledge of LGMD2A, AAV vectors, regulatory cassettes and mouse muscle testing to create this gene therapy for LGMD2A.

项目摘要 我们旨在开发一种用于肢体腰围肌肉营养不良2A型（LGMD2A）的基因疗法，一种常染色体隐性膜片 （AR）由于CAPN3突变引起的肌肉浪费障碍。 LGMD2A被认为是最普遍的 AR LGMD，但目前尚无治疗治疗的患者，他们通常依赖轮椅 诊断后的十年。我们和其他人表明，可以在 没有毒性的骨骼肌；这是使LGMD2A基因疗法的可行性成为现实的发现 目标;但是，LGMD2A在大多数其他LGMD中都是独一无二的，这一事实值得仔细开发基因 治疗载体。一个考虑因素是，由于心脏毒性，CAPN3无法在心脏中表达 Duchenne肌肉营养不良和其他LGMD并非如此。虽然有一些临床前的证明 概念研究已成功完成了小鼠的AAV-CAPN3过表达，没有一个 对具有LGMD2A的人类的任何基因疗法构建体的系统优化，尤其是一种考虑的基因疗法 相对骨骼肌肉与心脏基因表达问题。此外，因为LGMD2A优先 影响较慢的纤维，至关重要的是，最终进行临床试验的治疗结构是 优化用于缓慢的纤维表达。因为每个患者只能服用一次，所以必须 用于基因输送的构造是理想的。在此应用程序中，我们将使用一种迭代和系统的方法来 优化这些向量。我们组建了一支专家，合作团队，在该领域具有丰富的经验 调节盒，AAV载体发展和LGMD2A发病机理和易钙疗病动物模型。 该团队将共同努力，生成和测试一系列AAV向量的安全性和功效，以开发一个 LGMD2A的处理。这些向量将优化用于缓慢的纤维表达，同时避免心脏毒性。 一位团队成员Hauschka博士在很大程度上归功于创建绝大多数监管盒 用于目前用于Duchenne肌肉营养不良的AAV-GENE治疗试验。张伯伦博士设计的 优化了第一个微型和迷你肌营养不良蛋白，这构成了当前基因治疗中所有构建体的基础 DMD的临床试验。博士。 Spencer和Kramerova生成了许多导致该模型的鼠标模型 鉴定结果指标可用于测试LGMD2A的治疗干预措施。坎农博士是一个 国际知名的肌肉生理学家将对收缩功能进行生理评估。 该团队将应用他们对LGMD2A，AAV矢量，监管盒的广泛和累积知识 和小鼠肌肉测试为LGMD2A创建这种基因疗法。