Molecular tools for Bunyavirus antiviral screening

描述(由调查人员提供)：布尼亚病毒科的几种病毒是重要的人类病原体和潜在的生物恐怖主义病原体。针对其中一些病毒的疫苗确实存在，但除了利巴瓦林外，没有广谱抗病毒化合物能够抑制该家族中一个以上成员的复制。使用LaCrosse病毒(LAC)作为该家族的原型病毒，我们计划开发一种感染依赖和感染非感染相结合的筛选工具，并启动对具有抗本亚病毒活性的化合物的筛选。筛选工具将基于由噬菌体T7 RNA聚合酶或哺乳动物RNA聚合酶I产生的病毒启动子驱动的报告基因结构。类似的系统已经开发出来，并用于其他负链病毒，包括呼吸道合胞病毒、埃博拉病毒和甲型流感病毒。我们计划在自动化的高通量筛选程序中使用LAC感染独立系统来识别小分子抗病毒化合物。表现出抗病毒活性的化合物将被测试对LAC感染细胞的有效性。然后，我们将使用病毒启动子驱动的报告基因评估活性化合物的广谱抗病毒活性，这些报告基因来自布尼亚病毒汉坦(NIAID A类试剂)、Sin Norbre(NIAID A类试剂)、裂谷热(NIAID A类试剂)和克里米亚刚果出血热(NIAID C类试剂)。鉴于LAC是NIAID C类药物，完成这项申请应该会鉴定出对一种或几种NIAID优先病原体有活性的抗病毒化合物。高通量筛选、快速的病毒特异性传染性读数和可用的BSL2/BSL3空间相结合，使该应用成为与其他NIAID优先病原体一起筛选广谱抗菌化合物的有吸引力的原型。

DESCRIPTION (provided by investigator): Several viruses in the family Bunyaviridae are significant human pathogens and potential agents of bioterrorism. Vaccines against some of these viruses do exist, but with the exception of ribavarin, there are no broad-spectrum antiviral compounds capable of inhibiting replication of more than one member of the family. Using LaCrosse virus (LAC) as a prototype virus for the family, we plan to develop a combination of infection-dependent and infection-independent screening tools and to initiate a screen for compounds with anti-bunyavirus activity. The screening tools will be based on virus promoter-driven reporter gene constructs generated by bacteriophage T7 RNA polymerase or mammalian RNA polymerase I. Similar systems have been developed and utilized for other negative strand viruses including respiratory syncytial virus, Ebola virus, and influenza A virus. We plan to use the LAC infection-independent system in an automated, high throughput screening program to identify small molecule antiviral compounds. Compounds that exhibit antiviral activity will be tested for effectiveness against LAC-infected cells. Active compounds will then be assessed for broad-spectrum antiviral activity using virus promoter driven reporter genes derived from the bunyaviruses Hantaan (NIAID category A agent), Sin Nombre (NIAID category A agent), Rift Valley Fever (NIAID Category A agent) and Crimean Congo Hemorhagic Fever (NIAID category C agent). Given that LAC is a NIAID Category C agent completion of this application should identify antiviral compounds active against one and perhaps several NIAID priority pathogens. The combination of high throughput screening, rapid virus specific readouts of infectivity and available BSL2/BSL3 space make this application an attractive prototype for screening of broad spectrum antimicrobial compounds with other NIAID priority pathogens.