Investigating hepatocyte signaling driven by host-pathogen interactions

研究宿主-病原体相互作用驱动的肝细胞信号传导

• 批准号: 8821942
• 负责人: Alexis Kaushansky
• 金额: $ 12.91万
• 依托单位: SEATTLE BIOMEDICAL RESEARCH INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-05-08 至 2016-02-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8821942
• 关键词: Address; Apoptosis; Award; Binding; Biochemical; Biology; Cell Cycle Arrest; Cells; Complex; Data Set; Dependence; Development; Ensure; Environment; Evolution; Focus Groups; Goals; Hepatocyte; Host Defense; Human; Infection; Invaded; Lead; Life Cycle Stages; Liver; Malaria; Malignant - descriptor; Mediating; Membrane; Methods; Microarray Analysis; Molds; Molecular; Molecular Motors; Monitor; Outcome; Parasites; Parasitology; Pathway interactions; Pharmaceutical Preparations; Plasmodium; Plasmodium falciparum; Play; Population; Positioning Attribute; Post-Translational Protein Processing; Protein p53; Protein-Protein Interaction Map; Proteins; Research; Rodent; Role; Scientist; Signal Transduction; Signaling Protein; Staging; Toxoplasma; Toxoplasma gondii; Training; Tropism; Vacuole; Variant; Work; career; human disease; insight; killings; next generation; novel strategies; pathogen; prevent; prophylactic; protein protein interaction; public health relevance; research study; response; small molecule; technology development; trend

DESCRIPTION (provided by applicant): My career goal is to establish an independent research group, which simultaneously makes discoveries regarding the ways pathogens manipulate their host cells and contributes to the training and inspiring of the next generation of scientists. My background in cellular signaling, technology development, protein/protein interactions and parasitology has uniquely positioned me to apply a variety of powerful approaches to address important questions about pathogens and how they interact with their host cells. I have begun these efforts during my postdoctoral work in Stefan Kappe's group focusing on the malaria parasite and its hepatocyte host. I intend to expand these approaches moving forward to study the similarities and differences between divergent pathogens. Receipt of this K99/R00 award will greatly aid in accomplishing the vital tasks required to establish a productive and sustainable research group, and allow me to fully pursue my goal of leading a group focused on the fundamental biology of host-parasite interactions. Interaction networks of intracellular pathogens and their host cells are complex and predicted to be adaptive in promoting pathogen survival and life cycle progression. In this proposal, we utilize two related apicomplexan parasites: Plasmodium parasites whose co-evolution with the hepatocyte milieu has enabled their survival and Toxoplasma parasites whose persistence has occurred without requiring hepatocyte infection. We have recently demonstrated that liver stage malaria parasites subtly mold their host cells by lowering levels of the tumor suppressor p53, and that reversing this perturbation genetically or with small molecules dramatically reduces the ability of the parasite to develop within the liver. In this proposal, we aim to pursue three main objectives. First, we plan to further elucidate the mechanisms governing the malaria parasites dependence on low host p53 for intracellular survival, and investigate whether or not this perturbation is als required for robust Toxoplasma intracellular replication. We will then investigate which parasite molecules interact with p53 and whether or not this interaction is critical for Plasmodium liver stage development. Finally, we propose to expand upon our already intriguing molecular dataset for Plasmodium-infected hepatocytes by extensively interrogating signaling proteins in hepatocytes in response to both rodent and human Plasmodium parasites as well as Toxoplasma parasites using protein lysate microarrays, an approach that allows us to monitor hundreds of protein and post- translational modification levels using lysates derived from ~10,000 infected-cells. Taken together, the experiments proposed here will facilitate a deeper understanding of the environment in which intracellular parasites thrive and reveal interaction nodes that could be targeted by prophylactic drugs.

描述（由申请人提供）：我的职业目标是建立一个独立的研究小组，同时发现病原体操纵宿主细胞的方式，并为下一代的培训和启发做出贡献。 科学家。我在细胞信号传导、技术开发、蛋白质/蛋白质相互作用和寄生虫学方面的背景使我能够应用各种强大的方法来解决有关病原体及其如何与宿主细胞相互作用的重要问题。我在 Stefan Kappe 小组从事博士后工作期间就开始了这些努力，重点研究疟疾寄生虫及其肝细胞宿主。我打算扩展这些方法，以研究不同病原体之间的异同。获得 K99/R00 奖项将极大地有助于完成建立一个富有成效和可持续的研究小组所需的重要任务，并使我能够充分实现领导一个专注于宿主与寄生虫相互作用的基础生物学的小组的目标。细胞内病原体与其宿主细胞的相互作用网络是复杂的，预计在促进病原体存活和生命周期进展方面具有适应性。在本提案中，我们利用两种相关的顶端复门寄生虫：与肝细胞环境共同进化使其能够生存的疟原虫寄生虫和无需肝细胞感染即可持续存在的弓形虫寄生虫。我们最近证明，肝期疟原虫通过降低肿瘤抑制因子 p53 的水平巧妙地塑造其宿主细胞，并且通过基因或小分子逆转这种扰动可显着降低寄生虫在肝脏内发育的能力。在本提案中，我们旨在实现三个主要目标。首先，我们计划进一步阐明控制疟原虫依赖低宿主 p53 进行细胞内生存的机制，并研究这种扰动是否也是弓形虫细胞内复制的必要条件。然后我们将研究哪些寄生虫分子与 p53 相互作用，以及这种相互作用是否对疟原虫肝脏阶段的发育至关重要。最后，我们建议使用蛋白裂解物微阵列广泛询问肝细胞中针对啮齿动物和人类疟原虫寄生虫以及弓形虫寄生虫的信号蛋白，从而扩展我们已经很有趣的疟原虫感染肝细胞分子数据集，这种方法使我们能够使用衍生的裂解物监测数百种蛋白质和翻译后修饰水平 来自约 10,000 个受感染的细胞。总而言之，这里提出的实验将有助于更深入地了解细胞内寄生虫繁殖的环境，并揭示预防性药物可以针对的相互作用节点。