Vector Control Strategy Through Inhibition of Aedes aegypti Midgut Proteases

 描述(申请人提供)：蚊媒疾病继续对全球人类健康产生重大影响。然而，随着登革热和基孔肯雅病毒以及生物媒介埃及伊蚊在美国的存在，当地开始观察到对人类健康的影响。登革热和基孔肯雅病毒都只利用人类作为宿主和放大宿主。这一点很重要，因为雌性Ae。埃及伊蚊需要从人类宿主那里获得血液食物，才能获得适当的营养，为淋巴营养循环提供燃料。不幸的是，没有疫苗或治疗方法可用于对抗登革热和基孔肯雅病毒，因此防止这些病毒传播的唯一战略是通过控制蚊子种群(媒介)。目前使用杀虫剂的媒介控制策略在减少蚊子数量和减缓病原体传播方面已被证明是有效的。然而，Ae有所增加。埃及伊蚊对现有杀虫剂具有抗药性，随着世界人口的增加、城市化和缺乏有效的蚊子控制战略，蚊子数量将继续增长。因此，我们可能会经历登革热和基孔肯雅热感染的更高发病率。因此，我们的策略是针对参与血粉蛋白消化的蛋白水解酶。雌性艾氏。埃及伊蚊依靠消化血粉蛋白释放的营养物质来成熟和生产卵子。然而，在我们验证这些中肠蛋白酶作为一种潜在的媒介控制策略之前，我们必须首先充分了解单个中肠酶是如何消化血粉蛋白的。我们的研究将促进对Ae的认识。AaET、AaSPVI、AaSPVII和AaLT四种含量最丰富的中肠蛋白酶的生化研究。Aim 1.1将重点优化重组蚊虫蛋白水解酶的可溶性表达和纯化，然后进行生化和动力学分析，以确定底物特异性和最佳活性条件(Aim 1.2)。为了更深入地了解酶原(非活性)蛋白水解酶的调节，Aim 1.3将重点介绍水解酶激活的机制。除了这些研究，我们还建议从Ae产生一个供血的中肠的全球蛋白分解签名。埃及伊蚊(目标2)。在蛋白质组学的同时，这种方法将有助于识别以前没有发现的其他蛋白水解酶。综上所述，这些研究结果将有助于为今后针对Ae的工作奠定基础。埃及伊蚊的中肠蛋白酶和可能导致新的媒介控制策略的发展。

 DESCRIPTION (provided by applicant): Mosquito-borne diseases continue to have a significant impact on human health worldwide. However, with the presence of the Dengue fever and Chikungunya viruses and the biological vector Aedes aegypti in the United States, local impacts on human health are starting to be observed. Both the Dengue fever and Chikungunya viruses exclusively utilize humans as reservoirs and amplification hosts. This is important because the female Ae. aegypti mosquito requires a blood meal from a human host in order to obtain the proper nutrients to fuel the gonotrophic cycle. Unfortunately, no vaccines or treatments are available to combat the Dengue and Chikungunya viruses, and so the only strategy to prevent the spread of these viruses is through mosquito population (vector) control. Current vector control strategies using insecticides have proven effective in reducing the mosquito population and slowing down pathogen transmission. However, there has been an increase in Ae. aegypti mosquito resistance to available insecticides and with increasing world population, urbanization, and the lack of effective mosquito control strategies, the mosquito population will continue to grow. As a result, we will likely experience higher incidences of Dengue and Chikungunya infections. Therefore, our strategy is to target the proteolytic enzymes involved in blood meal protein digestion. The female Ae. aegypti mosquito relies on nutrients released by the digestion of blood meal proteins for egg maturation and production. However, before we can validate these midgut proteases as a potential vector control strategy, we must first fully understand how individual midgut proteases digest blood meal proteins. Our research will advance the knowledge of Ae. aegypti midgut proteases by biochemically studying the four most abundant midgut proteases (AaET, AaSPVI, AaSPVII, and AaLT). Aim 1.1 will focus on optimizing the soluble expression and purification of recombinant mosquito proteases, followed by biochemical and kinetic analysis to determine the substrate specificities and optimal activity conditions (Aim 1.2). To gain further insight into zymogen (inactive) protease regulation, Aim 1.3 will focus on the mechanism of protease activation. In addition to these studies, we propose to generate a global proteolytic signature of blood fed midguts from the Ae. aegypti mosquito (Aim 2). In parallel with proteomics, this approach will help identify other proteases that have not been previously identified. Taken together, the results of these studies will help lay the foundation for future work in targeting Ae. aegypti midgut proteases and may lead to the development of a new vector control strategy.