AAV VECTOR DEVELOPMENT FOR LUNG SPECIFIC GENE THERAPY

用于肺部特异性基因治疗的 AAV 载体开发

• 批准号: 2771589
• 负责人: JAMES P TREMPE
• 金额: $ 14.32万
• 依托单位: UNIVERSITY OF TOLEDO HEALTH SCI CAMPUS
• 依托单位国家: 美国
• 财政年份: 1996
• 资助国家: 美国
• 起止时间: 1996-09-30 至 2000-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2771589
• 关键词: Adenoviridae; cystic fibrosis; gene expression; gene therapy; laboratory mouse; technology /technique development; transfection /expression vector

DESCRIPTION (Taken directly from the application) Cystic Fibrosis (CF) is the most common lethal genetic disease of Caucasians and is the result of a single gene autosomal recessive disorder. Hopes for curing CF are currently riding on experimental gene therapy procedures in which a normal CF gene is permanently introduced into the lungs of affected individuals. The human parvovirus, adeno-associated virus (AAV), shows significant promise as a gene therapy vector. AAV is naturally defective, nonpathogenic and relatively easily to manipulate. In the absence of helper virus infection, the wild-type form of the virus integrates its genome into the long arm of chromosome 19. AAV is capable of successful CF gene delivery to rabbit lungs. Buoyed by the very early successes of gene delivery in animal systems, a Phase I clinical trial to deliver the human cystic fibrosis transmembrane regulator (CFTR) cDNA into CF patient lungs is imminent. Although much is known about the biology of AAV when grown in tissue culture cells, very little is known about the potential effects (both good and bad) of AAV gene therapy vectors. Only a handful of reports have appeared in the literature describing in vivo gene delivery to animal models. A vast amount of information about AAV gene therapy vectors remains to be uncovered before this exciting vector system should be used on a wide-spread basis in humans. To further develop and characterize AAV vectors we will study transgene delivery to mouse lungs. The aspects that we will study include: efficiency and longevity of expression, targeted cells, pharmacological enhancement of expression, the use of various gene promoters, neonatal gene delivery, repeated vector delivery and host responses. We have recently found that the AAV Rep78 protein, which is believed to be essential in establishment of latency, is covalently associated with virion DNA. We will study the potential of this particle-associated Rep78 to mediate vector integration. These experiments will provide essential information regarding the safety and efficacy of AAV vectors and lead to more widespread use of this important vector system.

描述（直接取自应用程序） 囊性纤维化（CF）是白种人最常见的致死性遗传病 并且是单基因常染色体隐性遗传病的结果。 的希望 治疗CF目前依靠实验性基因治疗程序， 正常的CF基因被永久地引入受影响的肺中， 个体 人细小病毒，腺相关病毒（AAV），显示 作为基因治疗载体具有重要前景。 AAV是天然缺陷的， 无致病性且相对容易操作。 在没有帮手的情况下 当病毒感染时，病毒的野生型形式将其基因组整合到 19号染色体的长臂 AAV能够成功地将CF基因 输送到兔子肺中。 受到基因早期成功的鼓舞， 在动物系统中进行的I期临床试验， 将囊性纤维化跨膜调节因子（CFTR）cDNA导入CF患者肺中， 迫在眉睫 虽然我们对AAV的生物学有很多了解， 组织培养细胞，对潜在的影响知之甚少（两者都是 好的和坏的）。 只有少数报道 出现在描述体内基因递送到动物的文献中 模型 关于AAV基因治疗载体的大量信息仍然存在 在这个令人兴奋的矢量系统应该被用在一个 在人类中广泛传播。 进一步开发和表征AV 载体，我们将研究转基因传递到小鼠肺部。 的方面 我们将研究包括：表达的效率和寿命，有针对性的 细胞，药理学增强表达，各种基因的应用 启动子、新生儿基因递送、重复载体递送和宿主 应答 我们最近发现AAV Rep78蛋白， 被认为是建立潜伏期必不可少的，是共价的， 与病毒体DNA有关。 我们将研究这方面的潜力 颗粒相关的Rep78介导载体整合。 这些实验 将提供有关AAV安全性和有效性的基本信息 并导致更广泛地使用这一重要的载体系统。