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

基本信息

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
项目类别：
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

项目摘要

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，该小组同时发现病原体操纵其宿主细胞的方式，并为下一代的培训和鼓舞科学家。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唯一地定位了采用各种强大的方法来解决有关病原体及其与宿主细胞相互作用的重要问题。我在斯特凡·卡佩（Stefan Kappe）小组的博士后工作中开始了这些努力，重点是疟疾寄生虫及其肝细胞宿主。我打算扩大这些方法向前发展，以研究发散病原体之间的相似性和差异。获得此K99/R00奖的收到将极大地有助于完成建立富有成效和可持续的研究小组所需的重要任务，并让我完全追求我的目标，即领导着一个专注于宿主 - 寄生虫相互作用的基本生物学的小组。细胞内病原体及其宿主细胞的相互作用网络是复杂的，并且预测在促进病原体生存和生命周期进程方面具有适应性。在此提案中，我们利用了两种相关的Apicomplexan寄生虫：疟原虫与肝细胞环境共同进化，使它们的生存和弓形虫寄生虫能够持久发生，它们的持久性发生了而无需肝细胞感染。我们最近证明，肝阶段疟疾寄生虫通过降低肿瘤抑制剂p53的水平微妙地塑造其宿主细胞，并且从遗传上或小分子逆转这种扰动会大大降低寄生虫在肝脏内发展的能力。在此提案中，我们旨在实现三个主要目标。首先，我们计划进一步阐明管理疟疾寄生虫对低宿主p53的机制，以用于细胞内存活，并研究这种扰动是否是强大的毒性毒素内细胞内复制所需的ALS。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阶段发育至关重要。最后，我们建议通过广泛询问肝细胞中的信号传导蛋白来响应啮齿动物和人疟原虫的寄生虫，并使用蛋白质的蛋白质来驱动蛋白质，从而扩展了我们已经有趣的分子数据集，以广泛询问肝细胞中的信号传导蛋白，并使用蛋白质的蛋白质来监测蛋白质，并使用蛋白质的蛋白质来监测蛋白质，从而扩大了蛋白质，并使用蛋白质的蛋白质溶解数量，从而扩展了肝细胞中的信号传导蛋白。来自约10,000个感染细胞。综上所述，此处提出的实验将有助于更深入地了解细胞内寄生虫蓬勃发展并揭示可能由预防性药物靶向的相互作用淋巴结。