Hochdurchsatz-Analyse pathogener Eigenschaften Herpesvirus-8-kodierter Gene in Endothelzellen

卡波西肉瘤(KS)是一种血管增生性疾病。人类疱疹病毒8型(HHV-8)在KS中起病原学作用。到目前为止，还不清楚HHV-8的哪些基因可能触发KS的发生。在这个项目中，我们将系统地确定HHV-8编码的84个基因在原代培养的人内皮细胞中的作用。特别是影响细胞增殖、单核细胞黏附、迁移和凋亡的肿瘤相关功能。此外，每个基因的效果将与其他HHV-8基因以及原癌基因c-myc和ras一起测试。最后，整个方法将在受到炎性细胞因子、血管生成生长因子和HIV-1-Tat蛋白不同外部刺激的内皮细胞上进行。为了达到这一目标，阵列表达载体的反向转染法将被建立为一个高通量的转导系统，它可以并行地确定数百个基因的功能。这项研究将确定在内皮细胞中具有最高致瘤激活潜力的HHV-8基因。此外，我们的方法可能提供一个有用的平台技术来确定Schwerpenkt计划范围内任何一组病毒和细胞基因的血管病变功能。

Kaposi's sarcoma (KS) is an angioproliferative disease. Human herpesvirus-8 (HHV-8) has an etiologic role in KS. It is unknown, as yet, which genes of HHV-8 may trigger KS development. In this project we will systematically determine the effects of each of the 84 genes encoded by HHV-8 in primary human endothelial cell cultures. Particularly tumor relevant functions, affecting cell proliferation, adhesiveness for monocytes, migration and apoptosis will be investigated. In addition, the effect of each gene will be tested in combination with every other HHV-8 gene and with the proto-oncogenes c-myc and ras. Finally, the whole approach will be performed with endothelial cells that have been subjected to different external stimulations by inflammatory cytokines, angiogenic growth factors and the HIV-1-Tat protein. To this goal, reverse transfection of arrayed expression plasmids will be established as a high throughput transfection system which allows to determine the functions of hundreds of genes in parallel. This study will identify the genes of HHV-8 with the highest tumorigenic activation potential in endothelial cells. Moreover, our approach may provide a useful platform technology to determine the vasculopathogenic function of any set of viral and cellular genes in the scope of the Schwerpunktprogramm.