Understanding and Targeting Host Processes Essential to Plasmodium Infection

了解并针对疟原虫感染所必需的宿主过程

• 批准号: 10735130
• 负责人: Emily R Derbyshire
• 金额: $ 50.12万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-06-01 至 2028-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10735130
• 关键词: AQP9 gene; Address; Affect; Affinity; Binding; Biological; Biological Models; Biology; Blood; Cell membrane; Cells; Chemicals; Chemistry; Complex; Cysteine; Development; Disease; Drug Targeting; Drug resistance; Environment; Erythrocytes; Foundations; Future; Genes; Genetic; Genetic Engineering; Genetic Transcription; Glycerol; Goals; Growth; Hepatocyte; Hour; Human; Image; Imaging Device; Immunoelectron Microscopy; Infection; Integration Host Factors; Invaded; Knowledge; Label; Life Cycle Stages; Liver; Malaria; Mammalian Cell; Maps; Measures; Membrane; Modeling; Modification; Molecular; Movement; Nutrient; Parasites; Permeability; Plasmodium; Plasmodium falciparum; Plasmodium vivax; Process; Protein Dynamics; Proteins; Research; Resolution; Role; Route; System; Testing; Vacuole; Vesicle; Visualization; Water; Work; aquaporin 3; cell fixing; chemoproteomics; clinically relevant; combat; disorder control; drug development; functional genomics; genetic approach; global health; insight; live cell imaging; malaria infection; mutant; novel; novel therapeutic intervention; pathogen; prevent; programs; prophylactic; recruit; reverse genetics; screening; small molecule; solute; spatiotemporal; targeted treatment; timeline; tool; trafficking; transcriptomics; transmission process; vaccine development; water channel

Project Summary Malaria is an ongoing global health burden, and the spread of drug resistance threatens progress made to eradicate the disease. The liver stage of the Plasmodium lifecycle is a promising target for drug and vaccine development as its inhibition would prevent disease manifestation and transmission. Previously, we mapped transcriptional changes throughout Plasmodium infection of hepatocytes and identified host processes critical to parasite viability. The chemical inhibition and genetic disruption of specific host proteins, such as aquaporin 3 (AQP3), were found to hinder liver stage parasite development. In mammalian cells, AQP3 transports water, glycerol, and other small solutes across cell membranes. Interestingly, we demonstrated that AQP3 localizes to the parasitophorous vacuolar membrane (PVM), the interface between the host and pathogen, in multiple Plasmodium species and stages. Our goal is to better understand host-parasite dynamics that support infection by determining how and why Plasmodium repurposes AQP3. In Aim 1, we plan to elucidate the recruitment of AQP3 to the host-pathogen interface using live-cell imaging with chemical and genetic tools. We will delineate the dynamics of AQP3 interactions with the tubulovesicular network, a membranous system that extends from the PVM. We will further probe the role of known trafficking motifs and the host endomembrane system to understand how AQP3 associates to the PVM and observe this association at an ultrastructural level with immuno-electron microscopy. In Aim 2, we will investigate AQP3 function during Plasmodium infection using a suite of imaging tools combined with AQP3 mutants to identify molecules affected by the host protein. In Aim 3, we will develop AQP3-targeting chemical probes to explore protein dynamics in the Plasmodium liver stages, including human-infective P. vivax and P. falciparum, where genetic approaches are currently unavailable. Fragment-based probe discovery will be used to identify covalent AQP3-binding molecules to label and study AQP3 in cells. Together, this work will provide insights into AQP3 recruitment and function during Plasmodium infection, thereby uncovering mechanisms that may be ubiquitously used by Plasmodium parasites to hijack host proteins. Our small molecule approach offers a route to complete fundamental biological studies probing host- parasite dynamics throughout different stages of the Plasmodium lifecycle and lays a foundation for future host- targeting compounds to address malaria infections.

项目摘要 疟疾是一个持续的全球健康负担，耐药性的传播威胁着在防治疟疾方面取得的进展。 根除疾病疟原虫生命周期的肝脏阶段是药物和疫苗的一个有前途的靶点 因为它的抑制将防止疾病的表现和传播。之前，我们绘制了 在疟原虫感染肝细胞过程中的转录变化，并确定了对 寄生虫生存能力对特定宿主蛋白质的化学抑制和遗传破坏，如水通道蛋白3 （AQP 3），被发现阻碍肝脏阶段寄生虫的发展。在哺乳动物细胞中，AQP 3转运水， 甘油和其他小溶质穿过细胞膜。有趣的是，我们证明AQP 3定位于 寄主液泡膜（PVM）是寄主和病原体之间的界面，在多种情况下， 疟原虫的种类和阶段。我们的目标是更好地了解支持感染的宿主-寄生虫动力学 通过确定疟原虫如何以及为什么重新利用AQP 3。在目标1中，我们计划阐明 使用活细胞成像与化学和遗传工具的宿主-病原体界面的AQP 3。我们将描绘 AQP 3与小管泡状网络相互作用的动力学，小管泡状网络是一种从 PVM。我们将进一步探讨已知的运输基序和宿主内膜系统的作用， 了解AQP 3如何与PVM相关，并在超微结构水平观察这种相关性， 免疫电镜在目标2中，我们将使用免疫组织化学方法研究AQP 3在疟原虫感染过程中的功能。 一套成像工具与AQP 3突变体相结合，以识别受宿主蛋白影响的分子。在目标3中， 我们将开发AQP 3靶向化学探针，以探索疟原虫肝脏阶段的蛋白质动力学， 包括人类感染的间日疟原虫和恶性疟原虫，目前还没有遗传方法。 基于片段的探针发现将用于鉴定共价AQP 3结合分子以进行标记和研究。 AQP 3在细胞中的表达。总之，这项工作将为疟原虫期间AQP 3的招募和功能提供深入见解 感染，从而揭示可能被疟原虫寄生虫普遍用于劫持宿主的机制 proteins.我们的小分子方法提供了一种完成基础生物学研究的途径， 在疟原虫生命周期的不同阶段的寄生虫动态，并奠定了基础，为未来的主机， 靶向化合物来解决疟疾感染。