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

体内基因转移很可能成为研究生理和病理条件下基因及其产物的重要工具。新近开发的第三代腺病毒载体具有良好的安全性、低毒性和免疫原性，可有效转导体内和体外多种复制和非复制细胞类型，具有高滴度和36kb的外源DNA容量，允许多个表达盒的基因转移或包含大的调控序列。在这个项目中，我们计划开发一种基于这些新腺病毒载体的技术，这种技术可以用于将基因定向转移到特定类型的细胞中。这将通过取消自然腺病毒的趋向性，同时在病毒衣壳中引入新的配体来实现，从而赋予新的靶向特异性。对于原理验证实验，我们将针对已知在激活的内皮细胞上高表达的av？3整合素。与该计划的其他成员合作，我们计划表达几种转基因，包括显性阴性的血管内皮生长因子受体(KDR/Flk-1)突变体和金属蛋白酶抑制剂。靶向性和有效性将在适当的体外和体内模型中进行测试。

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.