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Harnessing midgut mitochondrial dynamics to enhance Anopheline mosquito fitness

利用中肠线粒体动力学增强按蚊的适应性

基本信息

批准号：
8881816
负责人：
Shirley Luckhart
金额：
$ 77.55万
依托单位：
UNIVERSITY OF CALIFORNIA AT DAVIS
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-07-14 至 2016-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8881816
关键词：
Anopheles Genus Autophagocytosis Bioenergetics Biogenesis Biology Blood Caenorhabditis elegans Candidate Disease Gene Caspase Cell Death Cell Death Process Cell Differentiation process Cell Maintenance Cell Survival Cells Cessation of life Competence Culicidae Development Drosophila genus Drosophila melanogaster Electron Transport Energy Supply Ensure Epithelial Equilibrium Gene Expression Gene Targeting Genes Genetic Engineering Goals Growth Factor Health Homeostasis Human Immune Immunity Individual Infection Insulin Insulin-Like Growth Factor I Invertebrates Life Longevity Malaria Mammals Mediating Mediator of activation protein Metabolism Midgut Mitochondria Mitogen-Activated Protein Kinases Modeling Mus Nematoda Organism Oxidative Phosphorylation PTEN gene Parasite resistance Parasites Pathway interactions Pattern Peptides Phenocopy Phenotype Phosphotransferases Physiology Plants Plasmodium Plasmodium falciparum Play Population Predisposition Process Production Proteins Recombinants Regulation Reproduction Resistance Role Signal Pathway Signal Transduction Signaling Protein Somatomedins Stem cells Sum Tissue Survival Tissues Transcript Transgenic Organisms Work base biological adaptation to stress feeding fitness inhibitor/antagonist novel novel strategies nutrient metabolism overexpression peptide A reproductive response stem cell biology stem cell differentiation stem cell division tool transmission process vector

项目摘要

DESCRIPTION (provided by applicant): The use of transgenic mosquitoes in an integrated malaria control strategy will require mosquitoes that block parasite development and remain competitive with wild mosquito populations. Manipulation of key signaling cascades that regulate both immunity and fitness, such as the insulin/insulin growth factor signaling (IIS) cascade, represent a novel approach to achieving this goal. In mosquitoes and other model invertebrates the midgut functions as a center for IIS control of immunity, lifespan, metabolism, and reproduction. The effects of midgut IIS are largely mediated through mitochondrial dynamics, defined as the sum of mitochondrial biogenesis and clearance of damaged mitochondria through mitophagy. In invertebrates and mammals, IIS- dependent mitochondrial dynamics and, hence, mitochondrial metabolism regulate a wide range of important physiologies, including epithelial barrier integrity, stem cell maintenance and differentiation, lifespan and immunity, indicating that this regulation is fundamental in living organisms. Through our work with Anopheles stephensi, we have discovered that manipulation of IIS in the midgut alters the critical balance of mitochondrial biogenesis and mitophagy, resulting in phenotypic changes to mosquito resistance to Plasmodium falciparum infection, as well as mosquito lifespan and reproduction. Thus, we propose that IIS-dependent mitochondrial dynamics control A. stephensi "midgut health," which underlies the effects of IIS on immunity, lifespan, metabolism, and reproduction. To define how IIS-dependent mitochondrial dynamics regulate these important phenotypes, we will use five distinct treatments (Akt transgenic (TG), PTEN TG, insulin-fed, IGF1-fed, and manipulation of A. stephensi insulin-like peptides (AsILPs)) to "push and pull" mitochondrial dynamics in the midgut. This will allow us to identify and manipulate specific gene targets downstream of the IIS cascade that retain malaria parasite resistance while concurrently enhancing midgut health and overall mosquito fitness. To accomplish this, we will first define how midgut IIS regulates mitochondrial biogenesis and clearance through mitophagy and the impacts of these processes on energy homeostasis, stem cell maintenance and differentiation, epithelial integrity, and cell death processes. Since our four exogenous treatments (Akt TG, PTEN TG, insulin-fed, IGF1-fed) impact AsILP transcript expression in the midgut in predictable patterns, AsILPs likely function as natural mediators of IIS-dependent midgut mitochondrial dynamics. As such, we will manipulate AsILPs to alter midgut mitochondrial dynamics and to further clarify this association with IIS- dependent control of P. falciparum infection. Based on associations of IIS-dependent mitochondrial dynamics with mosquito fitness and resistance to parasite infection, we will identify candidate genes that optimally control midgut mitochondrial dynamics to specifically enhance these phenotypes. To this end, we will overexpress or disrupt expression of candidate genes in the midgut using a variety of tools, with the ultimate goal of generating stably transformed, fit A. stephensi that ar completely resistant to P. falciparum infection.

描述(申请人提供)：在综合疟疾控制战略中使用转基因蚊子将需要阻止寄生虫发展并保持与野生蚊子种群竞争的蚊子。操纵调节免疫和健康的关键信号级联，如胰岛素/胰岛素生长因子信号级联(IIS)，代表了实现这一目标的新方法。在蚊子和其他模型无脊椎动物中，中肠作为IIS控制免疫、寿命、新陈代谢和繁殖的中心。中肠IIS的作用在很大程度上是通过线粒体动力学来调节的，线粒体动力学被定义为线粒体生物发生和通过有丝分裂清除受损线粒体的总和。在无脊椎动物和哺乳动物中，依赖于IIS的线粒体动力学，因此，线粒体代谢调节一系列重要的生理，包括上皮屏障的完整性，干细胞的维持和分化，寿命和免疫，表明这种调节在活的生物体中是基本的。通过我们对斯氏按蚊的研究，我们发现在中肠操纵IIS改变了线粒体生物发生和有丝分裂的关键平衡，导致了蚊子对恶性疟原虫感染的抵抗力以及蚊子的寿命和繁殖的表型变化。因此，我们认为依赖于IIS的线粒体动力学控制着斯氏沼虾的“中肠健康”，这是IIS对免疫、寿命、新陈代谢和生殖的影响的基础。为了确定依赖IIS的线粒体动力学如何调节这些重要的表型，我们将使用五种不同的处理(Akt转基因(TG)、PTEN TG、胰岛素喂养、IGF1喂养和操纵斯氏拟胰岛素肽(AsILPs))来“推和拉”中肠线粒体动力学。这将使我们能够识别和操纵IIS级联下游的特定基因靶标，这些基因靶标保留了疟疾寄生虫的抵抗力，同时提高了中肠健康和蚊子的整体适应能力。为了实现这一目标，我们将首先定义中肠IIS如何通过吞噬有丝分裂来调节线粒体的生物发生和清除，以及这些过程对能量稳态、干细胞维持和分化、上皮完整性和细胞死亡过程的影响。由于我们的四种外源处理(Akt TG、PTEN TG、胰岛素喂养、IGF1喂养)以可预测的模式影响AsILP在中肠的转录表达，AsILP可能作为IIS依赖的中肠线粒体动力学的自然中介发挥作用。因此，我们将操作AsILPs来改变中肠线粒体动力学，并进一步阐明这种联系与依赖IIS控制恶性疟原虫感染的关系。基于依赖于IIS的线粒体动力学与蚊子适合度和对寄生虫感染的抵抗力之间的关联，我们将寻找最佳控制中肠线粒体动力学的候选基因，以特异性地增强这些表型。为此，我们将使用各种工具在中肠过表达或干扰候选基因的表达，最终目标是产生稳定转化的、适合斯氏疟原虫的、完全抵抗恶性疟原虫感染的斯氏疟原虫。