Zielgerichtete Transduktion aktivierter Endothelzellen mit adenoviralen Vektoren großer DNA-Kapazität und Expression von angiostatischen Faktoren und Matrix Metalloproteinase (MMP) Inhibitoren (Targeted transduction of activated endothelial cells with hig

体内基因转移将有可能成为研究基因及其产物在生理和病理条件下的一个非常重要的工具。最近开发的第三代腺病毒载体具有有利的安全性特征，降低的毒性和免疫原性，在体内和体外有效地转导许多复制和非复制细胞类型，产生高滴度，并具有36 kb外源DNA容量的优点，允许多个表达盒的基因转移或包含大的调节序列。在这个项目中，我们计划开发一种基于这些新的腺病毒载体的技术，用于将靶向基因转移到特定的细胞类型中。这将通过消除天然腺病毒嗜性并同时将赋予新靶向特异性的新配体引入病毒衣壳来实现。对于原理验证实验，我们将靶向已知在活化的内皮细胞上高度表达的α v β 3整联蛋白。在与该项目的其他成员合作中，我们计划表达几种转基因，包括显性阴性VEGF受体（KDR/Flk-1）突变体和金属蛋白酶抑制剂。将在适当的体外和体内模型中测试靶向特异性和有效性。

In vivo gene transfer will likely become a very important tool to study genes and their products in physiological and pathological conditions. A recently developed third-generation adenoviral vector has favourable safety features, decreased toxicity and immunogenicity, transduces efficiently many replicating and non-replicating celltypes in vivo and in vitro, is produced to high titers and has the advantage of 36 kb capacity for foreign DNA, allowing gene transfer of multiple expression cassettes or the inclusion of large regulatory sequences. Within this program we plan to develop a technology that is based on these new adenoviral vectors and that ca be used for targeted gene transfer into specific cell types. This will be achieved by abolishing the natural adenoviral tropism and at the same time introducing new ligands into the viral capsid that confer new targeting specificities. For proof-of-principle experiments we will target the avß3 integrin that is known to be highly expressed on activated endothelial cells. In collaboration with other members of this program we plan to express several transgenes including a dominant-negative VEGF receptor (KDR/Flk-1) mutant and metalloproteinase inhibitors. Targeting specificities and efficacies will be tested in appropriate in vitro and in vivo models.