Vector Control Strategy Through Inhibition of Aedes aegypti Midgut Proteases

 描述（由申请人提供）：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出现，开始观察到对当地人类健康的影响。登革热和基孔肯雅病毒都只利用人类作为宿主和扩增宿主。这一点很重要，因为女性。埃及蚊子需要人类宿主的血粉，以获得适当的营养来为生殖营养循环提供能量。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策略是通过蚊子种群（媒介）控制。目前使用杀虫剂的病媒控制策略已被证明在减少蚊子数量和减缓病原体传播方面是有效的。然而，Ae有所增加。埃及蚊子对现有杀虫剂的抗药性，随着世界人口的增加、城市化以及缺乏有效的蚊子控制策略，蚊子数量将继续增长。因此，我们可能会经历更高的登革热和基孔肯雅感染率。因此，我们的策略是靶向参与血粉蛋白质消化的蛋白水解酶。女性的爱。埃及蚊依靠消化血粉蛋白质释放的营养物来使卵成熟和产生。然而，在我们能够验证这些中肠蛋白酶作为潜在的载体控制策略之前，我们必须首先充分了解单个中肠蛋白酶如何消化血粉蛋白。我们的研究将促进对Ae的认识。通过生物化学研究四种最丰富的中肠蛋白酶（AaET、AaSPVI、AaSPVII和AaLT），对埃及伊蚊中肠蛋白酶进行了研究。目标1.1将集中于优化重组蚊子蛋白酶的可溶性表达和纯化，随后通过生物化学和动力学分析来确定底物特异性和最佳活性条件（目标1.2）。为了进一步深入了解酶原（无活性）蛋白酶的调节，目标1.3将集中在蛋白酶激活的机制。除了这些研究，我们建议从Ae产生一个全球性的血液喂养中肠蛋白水解签名。埃及伊蚊（Aim 2）。与蛋白质组学平行，这种方法将有助于识别以前未被识别的其他蛋白酶。综上所述，这些研究的结果将有助于奠定基础，为未来的工作，在针对Ae。埃及伊蚊中肠蛋白酶，并可能导致一种新的载体控制策略的发展。

 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.