Midgut mitochondria as the key to fit, Plasmodium-resistant Anopheline mosquitoes

中肠线粒体是抵抗疟原虫按蚊的关键

基本信息

批准号：
9464889
负责人：
CECILIA GIULIVI
金额：
$ 51.54万
依托单位：
UNIVERSITY OF IDAHO
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：
2016-08-19 至 2018-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9464889
关键词：
Midgut mitochondria key fit Plasmodium

项目摘要

DESCRIPTION (provided by applicant): The use of transgenic mosquitoes in an integrated malaria control strategy will require mosquitoes that completely block parasite development, yet remain competitive with wild mosquito populations. Manipulation of key signaling cascades regulating both immunity and fitness represents a novel approach to achieving this goal. In mosquitoes and other model invertebrates, the midgut functions as a center for insulin/insulin growth factor signaling (IIS), which controls immunity, lifespan, metabolism, and reproduction. The effects of midgut IIS are largely mediated through mitochondrial dynamics and activity, defined as mitochondrial biogenesis, bioenergetics, and clearance of damaged mitochondria through mitophagy. In invertebrates and mammals, IIS- dependent mitochondrial dynamics and mitochondrial metabolism regulate a wide range of important physiologies, including epithelial barrier integrity, stem cell maintenance and differentiation, lifespan and immunity, indicating tha this regulation is fundamental in living organisms. Through our work with Anopheles stephensi, we discovered that manipulation of IIS in the midgut alters the critical balance of mitochondrial dynamics and activity, resulting in phenotypic changes in mosquito resistance to Plasmodium falciparum infection as well as to mosquito lifespan and reproduction. Thus, we propose that IIS-dependent mitochondrial dynamics and activity control "midgut health" in A. stephensi, which underlies the effects of IIS on immunity, lifespan, metabolism, and reproduction. To define how IIS-dependent mitochondrial function regulates these important phenotypes, we will use five distinct treatments - Akt overexpression (increased IIS), PTEN overexpression (decreased IIS), provisioning with human insulin or IGF1, and manipulation of endogenous A. stephensi insulin-like peptides (AsILPs) - to "push and pull" mitochondrial dynamics and activity in the midgut. Specifically, we will define how midgut IIS-dependent mitochondrial biogenesis, bioenergetics, oxidative phosphorylation, and mitophagy regulate stem cell maintenance and differentiation, epithelial integrity, and cell death processes to control fitness and Plasmodium resistance. From these studies, we will identify and manipulate specific gene targets that directly regulate mitochondrial function to retain parasite resistance while concurrently enhancing mosquito fitness. We will overexpress four of these candidate genes based on our observations and published studies and two candidate genes identified from Aims 1 and 2 in the midgut singly or in pairs. Our goal for this project is to generate highly fit, P. falciparum resistant A. stephensi that can be deployed for malaria control. In the longer term, this transgenic platform could also be additive with other gene drivers and "customized" with anti-parasite effectors for sustainable resistance management.

描述（由申请人提供）：在综合疟疾控制策略中使用转基因蚊子将需要完全阻止寄生虫发育的蚊子，同时仍能与野生蚊子种群保持竞争性。调控免疫和适应性的关键信号级联代表了一种新的方法。在蚊子和其他模型无脊椎动物中，中肠充当胰岛素/胰岛素生长因子信号传导（IIS）的中心，其免疫控制、寿命、新陈代谢和繁殖的影响。中肠 IIS 的调节很大程度上是通过线粒体动力学和活性介导的，线粒体动力学和活性被定义为线粒体生物发生、生物能量学和通过线粒体自噬清除受损线粒体。在无脊椎动物和哺乳动物中，IIS 依赖性线粒体动力学和线粒体代谢调节广泛的重要生理机能，包括上皮细胞。屏障完整性、干细胞维持和分化、寿命和免疫，表明这种调节是生物体的基础。通过我们与斯氏按蚊的合作，我们发现了这种操纵。中肠中的 IIS 改变了线粒体动力学和活性的关键平衡，导致蚊子对恶性疟原虫感染的抵抗力以及蚊子的寿命和繁殖发生表型变化，因此，我们提出 IIS 依赖性线粒体动力学和活性控制“中肠”。 “在 A. stephensi 中，它是 IIS 对免疫、寿命、代谢和繁殖影响的基础。为了定义 IIS 依赖性线粒体功能如何调节这些重要表型，我们将使用五种不同的治疗方法 - Akt 过表达（增加 IIS）、PTEN 过表达（减少 IIS）、提供人胰岛素或 IGF1 以及操纵内源性斯蒂芬斯胰岛素样肽 (AsILP) - 来“推拉”线粒体动力学具体来说，我们将定义中肠 IIS 依赖性线粒体生物合成、生物能学、氧化磷酸化和中肠的活性。线粒体自噬调节干细胞的维持和分化、上皮完整性和细胞死亡过程，以控制适应性和疟原虫抗性。从这些研究中，我们将识别和操纵直接调节线粒体功能的特定基因靶标，以保持寄生虫抵抗力，同时增强蚊子的适应性。根据我们的高度观察和已发表的研究，以及从中肠中单独或成对地从目标 1 和 2 中识别出的两个候选基因，我们将过度表达这些候选基因中的四个。抗恶性疟原虫斯蒂芬氏疟原虫从长远来看，该转基因平台还可以与其他基因驱动器相结合，并通过抗寄生虫效应器进行“定制”，以实现可持续的耐药性管理。