Zytokin-vermitteltes Targeting von Adenovirusvektoren zur effizienten Transduktion hematopoietischer Zellen

腺病毒(Ad)载体打靶的目的是将基因有效地转移到特定类型的细胞和组织中。到目前为止，针对目标的努力收效甚微。最近，我们开发了一种新型的基因化学靶向平台，它允许灵活使用不同的配体，甚至与聚合物屏蔽相结合，以产生真正的重定向Ad颗粒。然而，我们的数据表明，不仅配体的类型，而且它在衣壳上的位置强烈地影响载体颗粒在细胞内的命运，从而影响转导效率。这可能是由于受体后在颗粒分解和/或颗粒运输水平上的限制。我们将使用基因化学靶向平台在分子和细胞生物学水平上表征Ad载体的受体后限制。我们将分析三种不同细胞内命运的配体：转铁蛋白、表皮生长因子和甘露聚糖对配体位置和配体拷贝数的影响。此外，还将表征屏蔽聚合物对Ad载体颗粒受体后命运的影响。此外，我们将研究生物响应键在将配体和屏蔽聚合物连接到载体颗粒时对受体后命运的影响。在分子水平上的详细分析可能形成合理设计靶向Ad载体的基础。所获得的结果可能会被转移到其他基因传递系统中。

Adenovirus (Ad) vector targeting aims for the efficient gene transfer to specific cell types and tissues. So far, targeting efforts have met with limited success. Recently, we developed a novel geneti-chemical targeting platform that allows for the flexible use of different ligands even in combination with polymer shielding for the generation of truly retargeted Ad particles. However, our data indicate that not only the type of ligand, but also its position on the capsid strongly influences the intracellular fate of the vector particle and, therefore, transduction efficiency. This is probably due to post-receptor restrictions at the level of particle disassembly and/or particle trafficking. We will use the geneti-chemical targeting platform to characterize post-receptor restrictions for Ad vectors at the molecular and cell biology level. We will analyze the influence of ligand positioning and ligand copy number for three ligands with different intracellular fates: transferrin, EGF, and mannans. In addition, the influence of shielding polymers on the post-receptor fate of Ad vector particles will be characterized. Further, we will investigate the effect of bioresponsive bonds on post-receptor fate when used to link ligands and shielding polymers to the vector particles. Detailed analysis at a molecular level may form a basis for rational design of targeted Ad vectors. The results obtained may likely be transferable to other gene delivery systems.