IMPROVED VIRAL VECTORS FOR GENE TRANSFER TO MUSCLE

改进的病毒载体用于基因转移到肌肉

• 批准号: 7404519
• 负责人: JEFFREY S CHAMBERLAIN
• 金额: $ 41.1万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-05-01 至 2008-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7404519
• 关键词: Adenovirus Vector; Affect; Age; Aging; Animals; Cardiovascular system; Cell Line; Cells; Clinical; Condition; Coupled; Development; Disease; Duchenne muscular dystrophy; Dystrophin; Elderly; Fiber; Gene Delivery; Gene Expression; Gene Transfer; Generations; Genes; Goals; Growth; Half-Life; Health; Helper Viruses; Hereditary Disease; Human; Individual; Insulin-Like Growth Factor I; Lead; Length; Mechanics; Mediating; Methods; Modeling; Mononuclear; Mus; Muscle; Muscle Weakness; Muscular Dystrophies; Myocardium; Myopathy; Numbers; Pathology; Patients; Personal Satisfaction; Preparation; Property; Protein Isoforms; Proteins; Relative (related person); Reporter Genes; Research Personnel; Sarcolemma; Serotyping; Signal Transduction; Skeletal Muscle; Skeletal system; Striated Muscles; System; Technology; Testing; Therapeutic; Toxic effect; Utrophin; Viral; Viral Vector; Work; adeno-associated viral vector; age related; combinatorial; experience; gene therapy; gutted adenoviral vector; improved; in vivo; mdx mouse; member; micro-dystrophin; model development; mouse model; prevent; programs; promoter; vector; young adult

The goals of this project are to develop and test improved viral vector systems for gene transfer to striated muscle. A particular focus is on the ability to transfer genes to muscles of adult and old mice, with an emphasis on models for Duchenne muscular dystrophy (DMD). DMD is among the most common human genetic diseases, and represents the most common genetic disease affecting skeletal muscle. Two vector systems will be explored: gutted adenoviral (Ad) and adeno-associated viral (AAV) vectors. Both of these systems have significant potential for use in gene transfer, but they each require additional development to enable clinical use in a safe and efficacious manner. Technological improvements to these vectors systems will therefore constitute a major component of the project goals. A second component of the project will explore the use of these vector technologies to modify muscles of adult and old animals, with a goal of correcting functional deficits that progressively increase with aging in both normal and dystrophic muscles. We will develop improved packaging cell lines, vectors and helper viruses to increase the yield, purity and ease of preparation of gutted Ad vectors. We will also explore the use of AAV serotype 6 for gene transfer to muscle. AAV serotype 6 displays highly efficient gene transfer to muscle, and we propose to optimize methods to obtain high titer stocks of highly purified AAV6 vectors displaying muscle-specific expression. Both types of vectors will be used to characterize the ability to prevent, halt or correct features of muscular dystrophy in the mdx mouse model of DMD by delivery of full-length, mini or micro-dystrophins. Studies will be conducted in young, adult and old mice, and will explore potential immunological consequences of delivery of these vectors to striated muscle. Finally, we will use these systems to explore additional phenotypic features of dystrophy in aging mice, with a goal of developing methods to arrest, and at least partially reverse, functional deficits of striated muscles by gene delivery of dystrophin and Igf-1. Studies that lead to improved methods for gene delivery and the amelioration of abnormalities of dystrophic muscles in adult and old animals will not only facilitate development of a treatment for DMD, but can also serve as a model for the development of genetic therapies for other genetic diseases and conditions associated with aging.

本项目的目标是开发和测试用于基因转移到横纹肌的改进的病毒载体系统。特别关注的是将基因转移到成年和老年小鼠肌肉中的能力，重点是杜氏肌营养不良症（DMD）模型。DMD是最常见的人类遗传性疾病之一，是影响骨骼肌的最常见遗传性疾病。将探索两种载体系统：内脏腺病毒（Ad）和腺相关病毒（AAV）载体。这两个系统都有巨大的潜力 用于基因转移，但它们各自需要额外的开发以使临床应用能够以安全有效的方式进行。因此，对这些病媒系统的技术改进将构成项目目标的一个主要组成部分。该项目的第二个组成部分将探索使用这些载体技术来修饰成年和老年动物的肌肉，目的是纠正随着正常和营养不良肌肉老化而逐渐增加的功能缺陷。我们将开发改进的包装细胞系、载体和辅助病毒，以提高产量、纯度和制备去内脏Ad载体的容易性。我们还将探索使用AAV血清型6将基因转移到肌肉中。AAV血清型6显示高效的基因转移到肌肉，我们建议优化方法，以获得高效价的股票高度纯化的AAV 6载体显示肌肉特异性表达。两种类型的载体将用于表征通过递送全长、小型或微型肌营养不良蛋白来预防、停止或纠正DMD的mdx小鼠模型中的肌营养不良特征的能力。研究将在年轻、成年和老年小鼠中进行，并将探索将这些载体递送至横纹肌的潜在免疫学后果。最后，我们将使用这些系统来探索衰老小鼠营养不良的其他表型特征， 通过肌营养不良蛋白和Igf-1的基因传递，开发阻止和至少部分逆转横纹肌功能缺陷的方法。导致改进基因递送方法和改善成年和老年动物营养不良肌肉异常的研究不仅将促进DMD治疗的开发，而且还可以作为开发与衰老相关的其他遗传疾病和病症的遗传疗法的模型。