How to starve a parasite: Manipulating CoA biosynthesis to control Plasmodium development in the mosquito

如何让寄生虫挨饿：操纵 CoA 生物合成来控制蚊子体内疟原虫的发育

• 批准号: 10656980
• 负责人: Shirley Luckhart
• 金额: $ 62.25万
• 依托单位: UNIVERSITY OF ARIZONA
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-04-14 至 2027-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10656980
• 关键词: Acetyl Coenzyme A; Affect; Anabolism; Anopheles Genus; Binding; Biology; Blood; CRISPR/Cas technology; Chemicals; Coenzyme A; Culicidae; Daphnia; Data; Development; Drosophila genus; Drug resistance; Engineering; Enzymes; Exhibits; Future; Generations; Genetic; Goals; Growth; Homology Modeling; Human; In Vitro; Infection; Inherited; Insecta; Insecticide Resistance; Lead; Libraries; Longevity; Malaria; Metabolism; Midgut; Molecular; Mosquito Control; Mosquito-borne infectious disease; Nutrient; Nutritional Requirements; Oocysts; PTEN gene; Pantothenate kinase; Pantothenic Acid; Parasite resistance; Parasitemia; Parasites; Parasitic infection; Pathway interactions; Pediatric Hospitals; Pharmaceutical Preparations; Pharmacotherapy; Phosphoric Monoester Hydrolases; Plasmodium; Plasmodium falciparum; Plasmodium yoelii; Protein Kinase; RNA Interference; Reproduction; Resistance; Resistance development; Resources; Saint Jude Children' s Research Hospital; Signal Transduction; Specificity; Starvation; Stress; Testing; Transgenic Organisms; Validation; Vectorial capacity; Vitamins; Work; analog; biological adaptation to stress; biological systems; cofactor; drug modification; fitness; genetic manipulation; human disease; humanized mouse; insight; kinase inhibitor; knock-down; life history; mouse model; novel; novel strategies; novel therapeutics; overexpression; prevent; resistant Plasmodium falciparum; screening; small molecule; success; tool; trait; uptake; vector mosquito

Project Summary Malaria parasites require pantothenate (Pan) from both the insect and mammalian hosts to synthesize coenzyme A (CoA) and acetyl-CoA (AC). Further, mosquito-stage parasites cannot take up preformed CoA from the insect host, so they are entirely dependent on mosquito Pan availability. Thus, we hypothesize that reducing Pan stores in the mosquito by increasing Pan kinase (PanK) activity and, in turn, CoA biosynthesis will limit parasite survival in the mosquito, without impacting the availability of CoA/AC to the mosquito itself. PanK is the rate-limiting enzyme in the CoA biosynthesis pathway and a logical target for our approach. In this study we will focus on increasing PanK activity in the mosquito to convert Pan into CoA and starve the malaria parasite of this essential precursor. To accomplish this we will utilize PanK-targeted small molecules or pantazines and genetic manipulation of PanK in our study host Anopheles stephensi. We will screen pantazines from a library of compounds developed by our collaborators at St. Jude Children’s Hospital. In Aim 1, we will use a Go-No Go strategy for pantazine screening that culminates in testing the capacity of selected pantazines to reduce P. falciparum and Plasmodium yoelii infections in A. stephensi. Aim 2 will validate the bioactivity and specificity of candidate pantazines identified in the screen in Aim 1. The specificity of candidate pantazines to activate PanK will be assessed through RNAi or CRISPR/Cas9 knockdown of PanK, followed by a characterization of the impact on Pan, CoA, AC and parasite infection success. Concurrent with Aims 1 and 2, we will generate transgenic A. stephensi with increased PanK activity and determine the impact on Pan levels and parasite survival in Aim 3. The generation of transgenic mosquitoes with increased midgut PanK expression will contribute to our assessment of PanK-dependent depletion of Pan stores on parasite infection as well as other aspects of mosquito biology related to vectorial capacity. Specifically, we will define the effects of mosquito PanK activation, via both pantazine treatment and molecular manipulations, on A. stephensi lifespan, stress responses, metabolism and reproduction. These studies will reveal important new insights into nutrient-driven mosquito-parasite interactions that drive parasite infection success and they will support future efforts to optimize pantazines and novel transgenic lines as distinct strategies for mosquito-targeted malaria control.

项目摘要 疟原虫需要昆虫和哺乳动物宿主的泛酸（Pan）来合成 辅酶A（CoA）和乙酰辅酶A（AC）。此外，蚊子阶段的寄生虫不能摄取预先形成的CoA 所以它们完全依赖于蚊盘的可用性。因此，我们假设， 通过增加Pan激酶（PanK）活性，进而增加CoA生物合成，减少Pan在蚊子中的储存 将限制寄生虫在蚊子中的存活，而不影响CoA/AC对蚊子本身的可用性。 PanK是CoA生物合成途径中的限速酶，也是我们方法的合理靶点。在这 在这项研究中，我们将专注于增加蚊子体内的PanK活性，将Pan转化为CoA，从而使疟疾挨饿。 这一基本前体的寄生虫。为了实现这一点，我们将利用PanK靶向小分子或 潘他嗪和PanK的遗传操作在我们的研究宿主斯氏按蚊。我们将筛选 pantazines从我们的合作者在圣裘德儿童医院开发的化合物库。在 目标1，我们将使用Go-No Go策略进行泛他嗪筛选，最终测试 选择潘达嗪以减少A.史蒂芬西目标2将 验证目标1中筛选鉴定的候选泛他嗪的生物活性和特异性。特异性 将通过RNAi或CRISPR/Cas9敲低来评估候选泛他嗪激活PanK的可能性。 PanK，然后描述对Pan、CoA、AC和寄生虫感染成功的影响。 与目的1和2同时，我们将产生转基因A.具有增加的PanK活性的stephensi， 确定目标3中对Pan水平和寄生虫存活的影响。转基因蚊子的产生 中肠PanK表达增加将有助于我们评估PanK依赖性的Pan耗竭， 储存寄生虫感染以及与媒介能力有关的蚊子生物学的其他方面。 具体来说，我们将通过泛他嗪治疗和分子生物学方法来确定蚊子PanK激活的效果。 操纵，在A. stephensi寿命，压力反应，新陈代谢和繁殖。这些研究将 揭示了对营养驱动的蚊子-寄生虫相互作用的重要新见解，这些相互作用驱动寄生虫感染 成功，他们将支持未来的努力，以优化泛他嗪和新的转基因品系， 以蚊子为目标的疟疾控制战略。