Understanding and Targeting Host Processes Essential to Plasmodium Infection

项目摘要 疟疾是一个持续的全球健康负担，耐药性的传播威胁到在以下方面取得的进展 根除这种疾病。疟原虫生命周期的肝脏阶段是药物和疫苗的一个有希望的目标 发展，因为它的抑制将防止疾病的表现和传播。之前，我们绘制了 疟原虫感染肝细胞的转录变化和识别的关键宿主过程 寄生虫的生存能力。特定宿主蛋白的化学抑制和遗传破坏，如水通道蛋白3 (AQP3)，被发现阻碍肝期寄生虫的发展。在哺乳动物细胞中，AQP3运输水， 甘油和其他跨越细胞膜的小溶质。有趣的是，我们演示了AQP3本地化到 寄生的液泡膜(PVM)是寄主和病原体之间的界面，在许多情况下 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-寄生虫的动态 通过确定疟原虫如何以及为什么改变AQP3的用途。在目标1中，我们计划阐明招募 使用带有化学和遗传工具的活细胞成像技术，将AQP3与宿主-病原体界面结合。我们将描绘出 AQP3与管泡网相互作用的动力学，管泡网是一个从 PVM。我们将进一步探讨已知的贩运主题和宿主内膜系统在 了解AQP3如何与PVM相关联，并在超微结构水平上观察这种关联 免疫电子显微镜。在目标2中，我们将研究AQP3在疟疾感染过程中的功能。 一套成像工具与AQP3突变体相结合，以识别受宿主蛋白影响的分子。在《目标3》中， 我们将开发针对AQP3的化学探针，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，这些地方目前还无法获得遗传方法。 基于片段的探针发现将用于识别共价AQP3结合分子以进行标记和研究 AQP3在细胞内表达。总之，这项工作将为了解AQP3在疟原虫中的招募和功能提供洞察力 感染，从而揭示了疟原虫可能无处不在地用来劫持宿主的机制 蛋白质。我们的小分子方法提供了一条完成基础生物学研究的途径，探测宿主- 寄生虫在疟原虫生命周期不同阶段的动态变化，为未来的宿主-- 以化合物为目标来应对疟疾感染。

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.