Developing Anti-viral Ribozymes to Suppress Arboviruses in Transgenic Mosquitoes

描述(申请人提供)：基孔肯雅热(CHIKV)、黄热病(YFV)和登革热(DENV)是每年导致数亿人感染和数十万人死亡的最麻烦的人类健康病原体。它们通过一种单一的蚊子物种埃及伊蚊传播给人类。目前还没有针对CHIK和DENV的有效人类疫苗，尽管有有效的疫苗可用，但YFV作为致命病原体重新出现，这表明持续根除的疫苗接种战略无效。对节肢动物媒介内孵化虫媒病毒的干扰正受到相当大的关注，因为这是一种中断病毒传播周期和降低传播给人类的效率的方法。一种这样的方法设想用那些抵抗感染或阻止疾病传播的蚊子来取代易受感染的蚊子。这种方法在环境安全、成本效益和长期疾病抑制方面具有明显的优势。转基因引入表达的抗病毒分子在蚊子载体中产生耐受性现在是一种真正的可能性。RIDL方法所反映的最新成功显示了巨大的前景，可以合理地预期与转基因耐受性相结合来在反弹蚊子种群中建立和维持转基因效应器。然而，这一途径必须与确保长期有效性和稳定的转基因表达相结合。我们的研究表明，核酶(RNA-酶)介导的病毒抑制可以提供一种有效的转基因免疫手段来对抗病毒。最近的工作证实了锤头状核酶和第一组内含子在抑制蚊子细胞中DENV感染方面的有效性。提出的这项研究试图在我们成功开发抗病毒核酶的基础上，将其作为对抗蚊子虫媒病毒感染的有效效应基因。特别是，这项建议将侧重于开发、优化和验证有效的核酶抑制策略，以对抗由同一蚊种传播的三种主要新出现和再次出现的疾病病原体，埃及伊蚊、DENV、CHIKV和YF。最终目标是能够提供有助于同时消除两种或更多此类病毒疾病的蚊子菌株。 公共卫生相关性：基孔肯雅热(CHIKV)、黄热病(YFV)和登革热(DENV)是最麻烦的单蚊子传播的人类健康病原体，每年导致数亿人感染和数十万人死亡。这项拟议的研究旨在开发抗病毒核酶，作为抑制这些虫媒病毒感染埃及伊蚊的手段，从而防止疾病传播。

DESCRIPTION (provided by applicant): Chikungunya (CHIKV), yellow fever (YFV), and Dengue (DENV) are among the most troublesome human health pathogens responsible for hundreds of millions of infections and hundreds of thousands of deaths each year. They are transmitted to human populations by a single mosquito species, Aedes aegypti. Effective human vaccines are not yet available for CHIK, and DENV, and the re-emergence of YFV as a lethal pathogen, in spite of the availability of an effective vaccine, suggests ineffectiveness of vaccination strategies for sustained eradication. Interference with the incubation arboviruses within the arthropod vector is receiving considerable attention as a way to interrupt the viral transmission cycle and reduce the efficiency of transmission to humans. One such approach envisions population replacement of susceptible mosquitoes with those resistant to infection or to halt disease transmission. This approach has distinct advantages of environmentally safety, cost effectiveness, and long-term disease suppression. Transgenic introduction of expressed anti-viral molecules to generate refractoriness in the mosquito vector is now a real possibility. Recent successes reflected in the RIDL approach show significant promise, and can reasonably be expected to be coupled with transgenic refractoriness to establish and maintain transgene effectors in rebound mosquito populations. However, this avenue must necessarily be coupled with ensuring both long term effectiveness and stable transgene expression. Our research is demonstrating that ribozyme (RNA-enzyme) mediated viral suppression can provide an effective means of transgenic immunization against viruses. Recent efforts have confirmed the effectiveness of hammerhead ribozymes and Group I introns in suppressing DENV infection in mosquito cells. The research proposed seeks to build upon our successes in developing anti-viral ribozymes as potent effector genes against arbovirus infections in mosquitoes. In particular, this proposal will focus on developing, optimizing, and validating effective ribozyme suppression strategies against three major emerging and re-emerging disease pathogens vectored by the same mosquito species, Aedes aegypti, DENV, CHIKV, and YF. The ultimate goal is to be able to provide mosquito strains that will be useful in simultaneously eliminating two or more of these viral diseases. PUBLIC HEALTH RELEVANCE: Chikungunya (CHIKV), yellow fever (YFV), and Dengue (DENV) are among the most troublesome single-mosquito transmitted human health pathogens causing hundreds of millions of infections and hundreds of thousands of deaths each year. This proposed research seeks to develop anti-viral ribozymes as means of suppressing the infection of these arbovirus in the mosquito, Aedes aegypti, thereby preventing disease transmission.