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Improving Anopheline fitness and resistance through fat body insulin signaling

通过脂肪体胰岛素信号改善按蚊健康和抵抗力

基本信息

批准号：
9293980
负责人：
Michael Allen Riehle
金额：
$ 22.54万
依托单位：
UNIVERSITY OF ARIZONA
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-06-10 至 2019-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9293980
关键词：
Affect Anopheles Genus Automobile Driving Cessation of life Culicidae Development Drosophila genus Drug resistance Egg Yolk Proteins Engineering Fat Body Fatty acid glycerol esters Fertility Generations Genes Genetic Engineering Glucose Glycogen Goals Growth Factor Hormones Human Immune Immune response Immune signaling Immunity Immunoblot Analysis Innate Immune Response Insecticide Resistance Insecticides Insulin Invertebrates Knock-out Link Lipids Longevity Malaria Metabolism Midgut Modeling Morbidity - disease rate Neurons Nutrient Organism Oxidative Stress PTEN gene Parasite resistance Parasites Pathway interactions Pattern Peptides Peripheral Pharmaceutical Preparations Phenotype Physiological Physiology Plasmodium Plasmodium falciparum Population Population Replacements Process Production Protein Biosynthesis RNA Interference Refractory Regulation Reproduction Research Resistance Role Salivary Glands Siblings Signal Transduction Solid Tissues Transcript Transgenic Organisms Translating Trehalose Vertebrates Work antimicrobial antimicrobial peptide assay development base comparative cost egg fitness improved insight insulin signaling killings knock-down malaria transmission mortality novel strategies nutrient metabolism offspring reproductive reproductive fitness senescence tool trait vector mosquito

项目摘要

Abstract Genetically modified mosquitoes resistant to Plasmodium development have been proposed as an alternative strategy to reduce malaria transmission. Several proof-of-principle studies have validated the idea that genetically modified mosquitoes can be refractory to the malaria parasite development, without incurring significant fitness loads. Recently, signaling cascades have been exploited to manipulate both parasite resistance and fitness. For example, manipulation of the insulin/insulin growth factor 1 signaling (IIS) pathway has been shown to confer Plasmodium resistance when up or down regulated via unique mechanisms, and extended lifespan when downregulated. However, the impact of IIS in other mosquito tissues has not been explored as extensively. The fat body of mosquitoes and other model invertebrates performs a number of functions, including storage of nutrients, production of yolk proteins and the synthesis of antimicrobial peptides. IIS has been implicated in these fat body processes leading to control of immunity, lifespan, metabolism, and reproduction. We previously generated a transgenic Anopheles stephensi line with increased insulin signaling in the peripheral fat body. Surprisingly, these transgenic mosquitoes survived significantly longer than their non-transgenic siblings, while in nearly every other organism and tissue increased IIS leads to a decrease in lifespan. To define how fat body IIS controls lifespan and to determine the impact fat body IIS has on reproduction, nutrient metabolism and Plasmodium resistance we will complete the following studies. Work in Drosophila suggest that fat body insulin signaling suppresses the expression of neuronal insulin-like peptides (ILPs) leading to increased lifespan. Thus, we will first examine transcript and peptide expression patterns of key AsILPs and knockdown putative AsILP targets via RNAi or Crispr knockout to validate the link between ILPs and lifespan extension. Our transgenic mosquito line also synthesized significantly more yolk protein than non-transgenic controls, although this did not translate into increased egg production during the first two gonotrophic cycles. Therefore, we will next conduct lifetime fecundity assays and development assays on the progeny to determine if an increase in lifetime reproductive fitness occurs. Third, due to the critical and well established role of IIS and the fat body on nutrient metabolism we will also quantify lipid, glycogen, glucose and trehalose levels at various physiological stages. Finally, the fat body is a key immune tissue regulating the production of anti-microbial peptides. As such we will assess the expression patterns of key immune genes and challenge transgenic mosquitoes with the most important human malaria parasite, Plasmodium falciparum. By the end of this project we will have a solid understanding of how fat body insulin signaling affects a range of physiologies impacting mosquito fitness and parasite resistance and will have developed new tools to generate highly fit Anopheles stephensi mosquitoes resistant to Plasmodium falciparum parasites.

摘要人们提出了一种替代办法，即用转基因蚊子来抵抗疟原虫的发展减少疟疾传播战略。一些原理验证研究证实了这一观点，转基因蚊子对疟疾寄生虫的发展具有抗性，重要的健身负荷。最近，信号级联被用来操纵这两种寄生虫抵抗力和适应性。例如，操纵胰岛素/胰岛素生长因子1信号传导（IIS）途径已显示当通过独特机制上调或下调时赋予疟原虫抗性，寿命延长。然而，IIS在其他蚊子组织中的影响尚未得到证实。探索得如此广泛。蚊子和其他模式无脊椎动物的脂肪体执行许多卵黄的主要功能包括营养物质的储存、卵黄蛋白的产生和抗菌肽的合成。 IIS与这些脂肪体过程有关，导致免疫力、寿命、代谢和生殖我们以前产生了一个转基因斯氏按蚊线增加胰岛素信号在周围脂肪体中。令人惊讶的是，这些转基因蚊子的存活时间比它们的在非转基因同胞中，而在几乎所有其他生物体和组织中，IIS的增加导致寿命定义脂肪体IIS如何控制寿命，并确定脂肪体IIS对生殖、营养代谢和疟原虫抗性，我们将完成以下研究。工作果蝇提示脂肪体胰岛素信号抑制神经元胰岛素样肽的表达（ILP）导致寿命延长。因此，我们将首先检查转录本和肽表达模式，关键AsILP和通过RNAi或Crispr敲除敲除推定的AsILP靶点，以验证AsILP之间的联系 ILP和寿命延长。我们的转基因蚊子系也合成了明显多于非转基因对照，尽管这并没有转化为前两个阶段的产蛋量增加生殖营养循环因此，我们接下来将对这两个物种进行终身生殖力测定和发育测定。后代，以确定是否增加终身生殖健康发生。第三，由于关键和良好的 IIS和脂肪体在营养代谢中的既定作用，我们还将量化脂质、糖原、葡萄糖和不同生理阶段的海藻糖水平。最后，脂肪体是一个关键的免疫组织，抗菌肽的生产。因此，我们将评估关键免疫基因的表达模式用最重要的人类疟疾寄生虫--恶性疟原虫来攻击转基因蚊子。通过这个项目的结束，我们将有一个坚实的了解脂肪体胰岛素信号如何影响一系列的，影响蚊子健康和寄生虫抵抗力的生理学，并将开发新的工具，斯氏按蚊对恶性疟原虫有抵抗力。