DEVELOPMENT OF AN AAV CFTR VECTOR PRODUCTION SYSTEM

该项目的长期目标是开发一个可扩展的GMP 腺相关病毒（AAV）的生产方法 用于治疗囊性纤维化的载体。 AAV-CFTR载体构建体 将被引入到各种细胞系中，并对其进行评估。 在添加必要的 包装功能。 将对许多包装构建体进行分析 在含有载体的细胞系中， 直接产生矢量粒子。 转导载体的纯化 粒子将通过细胞裂解、梯度洗脱和/或生物降解等物理方法进行优化。 离心和柱层析。 将使用AAV-CFTR载体 在治疗囊性纤维化的临床试验中， 纠正这种疾病的潜在病理生理学。 发展 这项技术可能会导致治疗的实质性改善， 这种疾病。 开发生产临床级的有效方法 AAV载体也可能导致用于治疗其他疾病的新产品。 除了囊性纤维化。 拟议的商业应用：开发AAV-CFTR载体， 囊性纤维化的治疗可能会产生一种基因递送"药物"， 可以减缓或阻止疾病的进展。 该产品将服务于 现有市场3万人。 此外还 设想AAV载体技术在其它基因中也是有用的， 转让申请，包括其他遗传疾病的治疗， 心血管疾病和癌症。

The long term objective of this project is to develop a scalable GMP manufacturing process for the production of adeno-associated virus (AAV) vectors for the treatment of cystic fibrosis. AAV-CFTR vector constructs will be introduced into a variety of cell lines and evaluated for their ability to produce vector particles following addition of requisite packaging functions. A number of packaging constructs will be assayed in the vector-containing cell lines for their ability to efficiently direct vector particle production. Purification of transducing vector particles will be optimized by physical methods of cell lysis, gradient centrifugation, and column chromatography. AAV-CFTR vectors will be used in clinical trials for the treatment of cystic fibrosis in an attempt to correct the underlying pathophysiology of this disease. Development of this technology could lead to substantive improvements in treatment of this disease. Development of efficient methods to produce clinical grade AAV vectors could also lead to new products for the treatment of other diseases in addition to cystic fibrosis. PROPOSED COMMERCIAL APPLICATION: Development of AAV-CFTR vectors for the treatment of cystic fibrosis could result in a gene delivery 'drug' that could slow or halt disease progression. This product would serve an existing market of 30,000 affected individuals. In addition, it is envisioned that AAV vector technology would be of utility in other gene transfer applications, including treatments for other genetic diseases, cardiovascular disease, and cancer.