IMPROVED ADENOVIRAL VECTORS FOR HEPATIC GENE THERAPY

描述：重组腺病毒载体为人类基因研究提供了可能性 治疗，因为它们有能力将许多组织 体内效率。对使用这些载体的热情一直是 受到针对病媒的强大免疫反应的影响 由于含有细胞低水平合成的载体衍生而来 抗原。腺病毒介导的基因转移在人类中持续存在 缺乏抗原依赖免疫的动物需要产生一种 抗原性载体较少。凯博士和他的同事们最近开发了一种 一种创建无载体的高滴度腺病毒载体的方法 负责产生免疫反应的基因组序列。 出乎意料的是，这些缺失的载体虽然在基因转移方面效率很高， 毒性最小或没有毒性，在体内不存在免疫活性或 免疫缺陷的动物。发生这种情况是因为这些删除的载体可能 在体外和体内，不能在转导细胞中复制它们的基因组。因此， 假设腺病毒DNA的持久性与其内在联系 它的复制能力。这项建议的主要目标是：1) 确定稳定和稳定所需的最低数量的腺病毒基因 将这些放回向量中，以生成可以持久保存的最小向量； 2)确定允许载体基因组持续存在的机制(S)；3) 测定急性毒性和抗原依赖性免疫反应 针对新载体；4)开发删除的载体系统以 产生一个整合的载体系统。这些研究的结果将 对基因的腺病毒的发展具有重要意义 心理治疗。

DESCRIPTION: Recombinant adenovirus vectors offer potential for human gene therapy because of their ability to transduce many tissues at high efficiency in vivo. The enthusiasm for use of these vectors has been tempered by a powerful immunologic response directed against vector containing cells because of the low level synthesis of vector derived antigens. The fact that adenovirus-mediated gene transfer is persistent in animals lacking antigen-dependent immunity establishes the need to produce a less antigenic vector. Dr. Kay and his colleagues recently developed a method for creating high titer adenovirus vectors that were devoid of vector genomic sequences responsible for producing the immune response. Unexpectedly, these deleted vectors, although efficient at gene transfer, with minimal or no toxicity, do not persist in vivo in immunocompetent or immunodeficient animals. This occurred because these deleted vectors could not replicate their genome in transduced cells in vitro and in vivo. Thus it is hypothesized that persistence of adenoviral DNA is inherently related to its ability to replicate. The major goals of this proposal are: 1) to determine the minimal number of adenovirus genes needed for stability and place these back into the vector to make a minimal vector that can persist; 2) determine the mechanism(s) allowing for vector genome persistence; 3) determine the acute toxicity and antigen-dependent immunological responses directed against the new vector; and 4) develop the deleted vector system to produce an integrating vector system. The results of these studies will have important implications for the development of adenoviruses for gene therapy.