Protection Against West Nile Virus by RNA Interference

通过 RNA 干扰预防西尼罗河病毒

• 批准号: 6882929
• 负责人: Raymond Allen Koski
• 金额: $ 35.19万
• 依托单位: L2 DIAGNOSTICS, LLC
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-05-01 至 2007-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6882929
• 关键词: West Nile virus; adeno associated virus group; animal morbidity; animal mortality; biotechnology; bioterrorism /chemical warfare; disease /disorder model; disease /disorder prevention /control; emerging infectious disease; encephalitis virus; gene delivery system; gene targeting; laboratory mouse; nonhuman therapy evaluation; small interfering RNA; therapy design /development; tissue /cell culture; transfection /expression vector; vaccine development; vaccine evaluation; vector vaccine; viral vaccines; virus load; virus replication

DESCRIPTION (provided by applicant): West Nile (WN) virus is a flavivirus that can cause fatal human encephalitis. Over the past four years, this virus has established itself across the United States. There are no approved vaccines or therapies to protect against WN virus. This project is designed to develop an effective anti-WN virus RNA interference therapy. Our hypothesis is that small interfering RNAs (siRNAs) targeting WN virus RNAs for degradation will inhibit viral replication and dissemination in vivo. Several siRNAs that inhibit WN virus in cultured cells will be identified in screening assays. The siRNAs showing robust interference in cellular assays will be evaluated in vivo using an established model of WN virus infection. The siRNAs will be expressed from adenovirus-associated virus vectors, including vectors designed to selectively target WN virus-infected cells. The AAV vectors will express siRNAs in mouse spleen, liver and brain, organs susceptible to WN virus. After WN virus challenge, the mice will be assessed for viral burden in different organs, and observed for morbidity, mortality. These experiments will provide the basis for Phase II experiments optimizing delivery of interfering RNAs to infected organs in animal models of WN virus infection.

描述（由申请方提供）：西尼罗河（WN）病毒是一种黄病毒，可导致致命的人类脑炎。在过去的四年里，这种病毒已经在美国各地建立了自己。目前还没有批准的疫苗或疗法来预防WN病毒。本项目旨在开发一种有效的抗WN病毒的RNA干扰疗法。我们的假设是，小干扰RNA（siRNA）靶向WN病毒RNA的降解将抑制病毒在体内的复制和传播。将在筛选测定中鉴定在培养细胞中抑制WN病毒的几种siRNA。将使用已建立的WN病毒感染模型在体内评价在细胞测定中显示出稳健干扰的siRNA。siRNA将从腺病毒相关病毒载体表达，包括设计用于选择性靶向WN病毒感染细胞的载体。AAV载体将在小鼠脾、肝和脑中表达siRNA，这些器官对WN病毒敏感。WN病毒攻击后，将评估小鼠不同器官中的病毒负荷，并观察发病率、死亡率。这些实验将为II期实验提供基础，以优化WN病毒感染动物模型中干扰RNA向感染器官的递送。