Improved Adenoviral Vectors for Hepatic Gene Therapy

描述(由申请人提供)： 重组腺病毒载体有过一段风雨飘摇的历史。尽管如此，基因缺失腺病毒的发展前景是光明的，因为它具有毒性低、基因转移效率高、DNA载量大等优点。该项目的目标是开发生产和评估保持异构体和/或整合到宿主染色体DNA中的基因缺失腺病毒载体所需的科学原理。新载体将在动物身上进行有效性和安全性评估，主要重点是肝脏基因转移。利用DNA转座子和位点特异性噬菌体整合酶，我们计划开发能够将表达盒整合到宿主染色体中的缺失载体。在血友病犬模型中，将比较上体型和整合型基因缺失载体在治疗性基因表达方面的持久性。这将使我们能够确定使用载体治疗遗传病的效用，在大多数情况下，遗传病需要终身基因表达。我们将开始尝试解开载体DNA的分子状态，并鉴定可能参与体内稳定外体型腺病毒载体DNA的细胞蛋白质。综上所述，这些研究将促进我们对与载体在RIVE中的持久性相关的载体-宿主相互作用的基本理解，以及促进临床前开发中的治疗应用。

DESCRIPTION (provided by applicant): Recombinant adenoviral vectors have had a stormy history. Nevertheless, there is promise in the development of gene-deleted adenoviruses because of reduced toxicity, efficient gene transfer, and large DNA carrying capacity. The goal of this project is to develop scientific principles required for production and evaluation of gene-deleted adenoviral vectors that remain episomal and/or integrate into host chromosomal DNA. The new vectors will be assessed in animals for efficacy as well as safety with the primary focus being on liver gene transfer. Using DNA transposons and site-specific phage integrases, we plan to develop gone-deleted vectors that can integrate an expression cassette into the host chromosome. Both episomal and integrating gene deleted vectors will be compared for longevity of therapeutic gene expression in a dog model of hemophilia. This will allow us to establish the utility of use of the vector for treating genetic diseases where life-long gene expression is required in most situations. We will begin to attempt to unravel the molecular state of the vector DNA and identify cellular proteins that may be involved in stabilizing episomal adenoviral vector DNAs in vivo. Taken together, these studies will advance our basic understanding of vector-host interactions related to persistence of vector in rive, as well as advancing therapeutic applications in preclinical development.