Indentification of Cellular Pathways Involvedin Rift Valley Fever Virus Infection

感染人类和家畜的流行性虫媒病毒疾病的出现导致了 世界范围内的发病率和死亡率。关于所需的宿主因素知之甚少， 这些病毒的复制周期，而不是限制发病机制的先天免疫途径， 阻碍了抗病毒治疗的发展。病毒感染相关细胞因子的鉴定 由于缺乏适合遗传学的病毒宿主系统， 筛选我们的主要研究目标是使用高通量RNAi和小分子筛选， 果蝇和哺乳动物细胞，以确定细胞和病毒因子的抑制剂， 致病机制，包括被病毒复制劫持的那些因素，以及被病毒复制所使用的那些系统。 抵抗病毒入侵的宿主我们的研究将集中在裂谷热病毒（RVFV， A类本扬病毒），但将通过MARGE内的合作扩展到其他本扬病毒， 医学意义RVFV由于其未满足的医疗和农业需求而受到特别关注，其 由美国农业部确定为可能的农业恐怖主义代理，试剂和实验的可用性 系统，并因为有机会与MARCE的成员合作。我们基于细胞的 使用可复制病毒的筛选方法能够鉴定化合物或基因， 针对病毒复制所需的任何病毒或细胞因子。此外，我们的方法具有 能够通过二级检测分析病毒复制来阐明这一要求的机制 生命周期中的每一步。同时应用基因组规模的RNAi筛选， RVFV和小分子筛选，均使用基于384孔板的高通量筛选测定 (HTS)我们所开发的技术，应该可以更快地识别抗病毒药物的细胞靶点， 这种医学上重要的病毒以及这种正在研究的病毒的其他成员的发展 家人

The emergence of epidemic arboviral diseases infecting both humans and domestic animals has led to significant world-wide morbidity and mortality. Little is known about the host factors required for the replication cycles of these viruses, and less about the innate immune pathways that restrict pathogenesis, impeding the development of antiviral treatments. The identification of cellular factors involved in viral replication and pathogenesis has been difficult due to the lack of virus-host systems amenable to genetic screening. Our primary research goal is to use a high-throughput RNAi and small molecule screens in Drosophila and mammalian cells to identify inhibitors of both cellular and viral factors that regulate pathogenesis, including both those factors hijacked by the virus for replication, and those systems used by the host to combat the viral invader. Our studies will be focused on Rift Valley Fever Virus (RVFV, a Category A bunyavirus), but will be extended via collaborations within MARGE to other bunyaviruses of medical significance. RVFV is of particular interest due to its unmet medical and agricultural need, its identification by the USDA as a likely agroterrorism agent, the availability of reagents and experimental systems, and because of the opportunity to collaborate with members of the MARCE. Our cell-based screening approach using replication competent virus enables the identification of compounds or genes that target any factor¿viral or cellular¿that is required for viral replication. Furthermore, our method has the ability to elucidate the mechanism of this requirement by dissecting viral replication with secondary assays against each step in the lifecycle. The simultaneous application of genome-scale RNAi screens against RVFV and small molecule screens, both using a 384 well plate-based high throughput screening assay (HTS) that we have developed, should lead to more rapid identification of cellular targets for antiviral drug development for this medically important virus, as well as for other members of this under-studied virus family.