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.

项目概要磷酸肌醇 (IP) 是可扩散的细胞内信使（称为“IP 代码”），执行关键的任务在真核细胞中发挥功能，但这些分子对免疫和宿主防御的贡献仍然存在通过公正的方法，我们的实验室最近发现了知识产权的惊人新作用。宿主-病毒界面的分子：（1）在坏死性凋亡过程中，促炎性细胞死亡机制至关重要抗病毒免疫和（2）人类鼻病毒（RV）感染期间，其中最常见的我们发现 IP 激酶活性可产生细胞 IP。特征（即 IP3、IP4、IP5、IP6）对于坏死性凋亡和 RV 感染都至关重要（通过明显不同的方式）我们证明IP分子可以直接控制坏死性刽子手，混合谱系。激酶样（MLKL），以释放坏死性细胞死亡，相反，我们发现 RV 感染依赖于 IP。细胞死亡之前的阶段并且独立于 MLKL，表明 IP 的宿主或病毒靶标尚未确定。然而，目前尚不清楚不同的IP如何在分子水平上控制这些过程，如何 IP 激酶精确协作以确定细胞 IP 代码，以及该代码在细胞分裂过程中的调节方式感染或细胞因子刺激介导免疫我们的中心假设是肌醇磷酸盐和它们的激酶在先天免疫反应和病毒感染过程中发挥重要的调节作用在这里，我们将确定与宿主或病毒靶标的相互作用的精确机制。通过使用强大的遗传和免疫过程，肌醇磷酸盐编码生化工具包，并将我们在分子遗传学和病毒学方面的专业知识与协作团队相结合具体来说，我们将 (1) 绘制遗传和生物化学图谱。在细胞和生化测定中 IP 物种与坏死性刽子手 MLKL 的物理相互作用， (2) 定义 IP 代码控制 RV 感染的分子机制。这些研究将为免疫防御调节和病毒发病机制提供基础见解。希望这项工作能够为理解 IP 代码在免疫和感染中的作用奠定基础生物学，并为开发增强抗微生物和抗微生物作用的新方法提供基础炎症治疗。