Molecular tools for Bunyavirus antiviral screening

描述（由研究者提供）：布尼亚病毒科中的几种病毒是重要的人类病原体和潜在的生物恐怖主义因子。针对这些病毒中的一些的疫苗确实存在，但是除了ribavarin之外，没有能够抑制家族中一个以上成员的复制的广谱抗病毒化合物。使用长曲棍球病毒（LAC）作为原型病毒的家庭，我们计划开发一个组合的感染依赖性和感染无关的筛选工具，并开始筛选具有抗布尼亚病毒活性的化合物。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构建体。已经开发了类似的系统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、高通量筛选程序中使用LAC感染无关系统来鉴定小分子抗病毒化合物。将测试表现出抗病毒活性的化合物对LAC感染的细胞的有效性。然后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，所述病毒启动子驱动的报告基因来源于布尼亚病毒汉坦（NIAID A类病原体）、Sin Nombre（NIAID A类病原体）、裂谷热（NIAID A类病原体）和克里米亚刚果出血热（NIAID C类病原体）。鉴于LAC是NIAID C类制剂，完成本申请应确定对一种或几种NIAID优先病原体具有活性的抗病毒化合物。高通量筛选、感染性的快速病毒特异性读数和可用的BSL 2/BSL 3空间的组合使该应用成为筛选具有其他NIAID优先病原体的广谱抗微生物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.