KINASE NETWORKS CONTROLLING FLAVIVIRUS REPLICATION

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 这个项目的目标是确定宿主细胞--黄病毒复制或抗黄病毒免疫反应所必需的途径。我们将通过使用系统的化学基因组学方法来识别调节黄病毒复制的宿主细胞激酶网络来识别这些途径。通过基于细胞的对登革病毒(DENV)复制抑制因子的激酶定向小分子文库的筛选，我们有望识别抑制DENV生命周期不同步骤的激酶抑制剂。在二次筛选中，我们将选择具有高选择性指数、广泛的细胞类型特异性和抗其他黄病毒(WNV、JEV、YFV)活性的化合物。通过结合计算、功能基因组学和生物化学方法，可以确定特定化合物所针对的激酶。因此确定的黄病毒复制的细胞途径的重要性将通过基因敲除给定宿主细胞途径的关键成分来评估。选定的化合物将使用西尼罗河病毒动物模型在体内进行测试。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. This goal of this project is to identify host cell-pathways essential for flavivirus replication or for anti-flavivirus immune responses. We will identify these pathways by using a systematic chemical genomics approach to identify host cell kinase networks that modulate flavivirus replication. By cell-based screening of kinase-directed small molecule libraries for inhibitors of Dengue-virus (DENV) replication, we expect to identify kinase inhibitors that inhibit different steps of the DENV life cycle. In secondary screens we will select compounds with a high selectivity index, broad cell type specificity and activity against other flaviviruses (WNV, JEV, YFV). The kinases targeted by a given compound will be identified by combining computational, functional genomics and biochemical approaches. The importance of cellular pathways thus identified for flavivirus replication will be evaluated by gene knockdown of key components of a given host cell pathway. Selected compounds will be tested in vivo using a West Nile Virus animal model.