Deciphering the inositol phosphate code in viral pathogenesis and immunity

破译病毒发病机制和免疫中的肌醇磷酸密码

基本信息

批准号：
10397756
负责人：
Jan E Carette
金额：
$ 7.56万
依托单位：
STANFORD UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-07-01 至 2023-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10397756
关键词：
Affect Affinity Amino Acids Anti-Inflammatory Agents Antiviral Agents Artificial Membranes Bacterial Infections Binding Biochemical Biochemical Genetics Biochemistry Biological Assay Biology CRISPR/Cas technology Cell Death Cells Cellular Assay Charge Code Collaborations Collection Development Diffuse Disease Drug Design Economic Burden Eukaryotic Cell Foundations Genetic Goals Host Defense Human Immune Immunity Immunomodulators In Vitro Infection Infectious Agent Inflammation Inflammatory Innate Immune Response Inositol Inositol Phosphates Knock-out Laboratories Life Cycle Stages Malignant Neoplasms Maps Mass Spectrum Analysis Mediating Mediator of activation protein Membrane Methods Molecular Molecular Genetics Molecular Virology Necrosis Neurodegenerative Disorders Pathogenesis Pathologic Phosphotransferases Play Process RNA replication Reagent Regulation Reporter Rhinovirus Rhinovirus infection Role Signal Pathway Testing Therapeutic Translations Viral Viral Pathogenesis Virus Virus Diseases Work antimicrobial antiviral immunity base cytokine genome-wide global health immunoregulation insight ischemic injury new therapeutic target novel response

项目摘要

PROJECT SUMMARY Inositol phosphates (IPs) are diffusible intracellular messengers (termed the “IP code”) that perform key functions in the eukaryotic cell, yet the contributions of these molecules to immunity and host defense remains underexplored. Through unbiased approaches, our laboratory has recently uncovered striking new roles for IP molecules at the host-virus interface: (1) during necroptosis, a pro-inflammatory cell death mechanism critical for antiviral immunity and (2) during infection with human rhinoviruses (RV), among the most common infectious agents in human beings. We have found that IP kinase activity that produces the cellular IP signature (i.e., IP3, IP4, IP5, IP6) is critical for both necroptosis and RV infection via apparently distinct mechanisms. We showed that IP molecules can directly control the necroptotic executioner, mixed-lineage kinase like (MLKL), to unleash necrotic cell death. In contrast, we found that RV infection depends on IPs at a stage preceding cell death and independent of MLKL, indicating as yet unidentified host or viral targets for IPs. However, it is currently unknown how the different IPs act to control these processes on a molecular level, how IP kinases precisely collaborate to determine the cellular IP code, and how this code is regulated during infection or cytokine stimulation to mediate immunity. Our central hypothesis is that inositol phosphates and their kinases play important regulatory roles in innate immune responses and during viral infection through interactions with host or viral targets. Here, we will determine the precise mechanisms for engagement of the inositol phosphate code by these fundamental immune processes by employing a robust genetic and biochemical toolkit and combining our expertise in molecular genetics and virology with a collaborative team of leading experts in inositol phosphate biology and biochemistry. Specifically, we will (1) map the genetic and physical interactions of IP species with the necroptotic executioner MLKL in cellular and biochemical assays, and (2) define the molecular mechanism(s) governing regulation of RV infection by the IP code. Together, these studies will provide fundamental insight into regulation of immune defenses and viral pathogenesis. We expect this work to form a foundation for understanding the roles of the IP code in immunity and infection biology and to provide a basis for development of novel methods to enhance anti-microbial and anti- inflammatory therapeutics.

项目摘要肌醇磷酸盐（IPS）是可扩散的细胞内信使（称为“ IP代码”）真核细胞的功能，但这些分子对免疫和宿主防御的贡献仍然存在尚未得到充实的。通过公正的方法，我们的实验室最近发现了IP的新角色宿主病毒界面的分子：（1）在坏死性局势，促炎细胞死亡机制临界对于最常见的人类鼻病毒（RV）感染期间的抗病毒免疫力和（2）人类中的传染性药物。我们发现产生细胞IP的IP激酶活性签名（即IP3，IP4，IP5，IP6）对于坏死性和RV感染至关重要机制。我们表明，IP分子可以直接控制坏死式executionuter，混合细节类似（MLKL）的激酶，释放坏死细胞死亡。相反，我们发现RV感染取决于A 在细胞死亡之前的阶段，独立于MLKL，表明IPS的宿主或病毒靶标尚不确定。但是，目前尚不清楚不同的IP如何在分子水平上控制这些过程，以及如何 IP激酶正精确地协作以确定蜂窝IP代码，以及如何调节此代码。感染或细胞因子刺激以介导免疫。我们的中心假设是肌醇磷酸盐和他们的激酶在先天免疫反应以及通过与宿主或病毒靶标的相互作用。在这里，我们将确定参与的确切机制通过使用强大的遗传和生化工具包，并将我们在分子遗传学和病毒学方面的专业知识与一个合作团队相结合肌醇磷酸盐生物学和生物化学领域的主要专家。具体而言，我们将（1）映射遗传和 IP物种与细胞和生化测定中的坏死otecution子手MLKL的物理相互作用，（2）定义通过IP代码对RV感染调节的分子机制。一起，这些研究将提供对免疫防御和病毒发病机理调节的基本见解。我们期望这项工作构成理解IP代码在免疫和感染中的作用的基础生物学并为开发新方法提供基础，以增强抗微生物和抗炎症疗法。