GENE MODIFICATION OF ADENOVIRUS CAPSID PROTEINS

特定靶细胞的摄取是基因的关键第一步 转移，在大多数制度中，这受到内生进入的限制 特定基因转移载体/病毒所使用的途径。我们有 成功地将Ad5载体与靶细胞结合，通过 一种不同于亚型通常使用的高亲和力受体- C组病毒。尽管我们在这方面取得了相当大的进展 在这方面，我们仍有相当大的需要继续努力 目标广告载体。如果我们要以组织为目标，这一点尤其正确 例如已经报道过高分化的呼吸道上皮 缺乏高亲和力和低亲和力的受体 被C亚群病毒用来进入。 腺病毒介导的基因转移的基本步骤：1)附着于细胞 通过高亲和力受体(与CAR结合的纤维，MHC-1)，2) 由五色子与细胞相互作用介导的促进内化 αvbeta3，5整合素，以及3)内体逃逸，这是 主要衣壳蛋白Hexon的构象变化是 Ad载体使其成为最有效的基因转移的几个方面 矢量可用。这些蛋白质也是先天的主要靶标。 和获得性免疫系统，这是缺乏 腺病毒载体基因转移的持久性和中和性 损害重复给药有效性的豁免权 广告载体。 这个项目的重点是对主要衣壳进行基因改造 腺病毒蛋白(Ad)：六邻体、纤维和五子体占优势 基因转移到呼吸道上皮细胞的基因转移。这是我们的 认为这些蛋白质是阳性和阴性的关键介质 Ad基因转移载体的负面属性和我们的能力 通过基因操纵来操纵它们将导致 更有效的基因转移，更大程度的靶细胞 特异性，最后是不纯性的下降。开发 我们载体中修饰衣壳蛋白的能力将直接 适用于任何广告载体系统，包括“无勇气载体”， 嵌合病毒载体和携带大DNA的Ad载体 分子。

Uptake by a specific target cell is a critical first step in gene transfer, which in most systems is limited by the endogenous entry pathway used by a particular gene transfer vector/virus. We have successfully engineered the Ad 5 vector to bind to a target cell through a different high affinity receptor than that normally used by the sub- group C viruses. Although we have made considerable progress in this area, there is still a considerable need for continuing our efforts to target Ad vectors. This is especially true if we are to target tissues such as well differentiated airway epithelium which have been reported to lack the high affinity as well as low affinity receptors that are used by subgroup C viruses for entry. Steps essential to Ad-mediated gene transfer: 1) attachment to the cell via the high affinity receptor (fiber binding to CAR, MHC-1), 2) facilitated internalization mediated by penton interaction with the alphavbeta3,5 integrins, and 3) endosomal escape which is a function of conformational changes in the major capsid protein hexon, are the aspects of Ad vectors which make them the most efficient gene transfer vector available. These proteins are also the primary targets of innate and acquired immune systems which are responsible for the lack of persistence of gene transfer by Ad vectors as well as the neutralizing immunity which compromises the effectiveness of repeat administration of Ad vectors. The focus of this project is to genetically modify the major capsid proteins of Adenovirus (Ad): hexon, fiber, and penton to the advantage of gene transfer of gene transfer to airway epithelial cells. It is our position that these proteins are the key mediators of both positive and negative attributes of Ad gene transfer vectors and our ability to genetically manipulate to genetically manipulate them will result in more efficient gene transfer, a greater degree of target cell specificity and finally a decreased in imunogenicity. Developing the capacity to modify the capsid proteins in our vectors will have direct application to any Ad vector system, including the "gutless vectors", chimeric virus vectors, and Ad vectors used to piggyback large DNA molecules.