Rhinovirus Pathogenesis and Host Range

鼻病毒发病机制和宿主范围

基本信息

批准号：
7893161
负责人：
VINCENT R RACANIELLO
金额：
$ 40.68万
依托单位：
COLUMBIA UNIVERSITY HEALTH SCIENCES
依托单位国家：
美国
项目类别：
财政年份：
2002
资助国家：
美国
起止时间：
2002-02-01 至 2012-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7893161
关键词：
Address Adult Amino Acids Animal Model Animal Viruses Antiviral Agents Architecture Asthma Biology CDPdiacylglycerol-inositol 3-phosphatidyltransferase Cell Culture Techniques Cells Cleaved cell Common Cold Complex Cultured Cells Cytoplasm DNA DNA Sequence Rearrangement Dendritic Cells Dominant-Negative Mutation Enzymes Family Genome Goals Golgi Apparatus Growth Human Human poliovirus Infection Interferons Internal Ribosome Entry Site Laboratories Lead Life Lung Lytic Mediating Medical Membrane Molecular Mus Neonatal Oral Poliovirus Vaccine Organ Pathogenesis Pattern Recognition Pattern recognition receptor Play Polioviruses Primates Protein Secretion Proteins RNA Viruses Reporter Research Research Personnel Respiratory System Respiratory tract structure Rhinovirus Role Signal Transduction Site Structure Surface TLR3 gene TLR7 gene Therapeutic Intervention Transgenic Mice Translations Tropism Vesicle Viral Viral Genome Viral Proteins Virus Virus Diseases attenuation base cytokine design human disease inhibitor/antagonist lipid biosynthesis macromolecule mouse model mutant pathogen poliovirus receptor prevent programs research study response sensor trafficking

项目摘要

DESCRIPTION (provided by applicant): Human rhinoviruses (HRVs) are major causative agents of the common cold, one of the most frequent human infections. They are also the most common pathogens associated with asthma exacerbations. Despite the medical importance of rhinoviruses, the pathogenesis of rhinovirus infection is poorly understood because a convenient animal model is not available. The experiments in this proposal address a wide range of experimental questions on the biology of rhinovirus, encompassing replication in cell culture, the molecular basis of host range, innate responses to infection, and establishing replication in mice. These goals will be accomplished through the following four specific aims. 1. Determine the role in rhinovirus replication of cell proteins that interact with the viral 2B and 3A proteins. We have shown that the 3A protein of HRV39 interacts with GCP60 and FinGERS, proteins involved in vesicle traffic, and with an enzyme involved in lipid biosynthesis, CDIPT (phosphatidylinositol synthase). Cell proteins that interact with 2B will also be identified. We will assess the role of these cell proteins in viral replication, alteration of membrane ultrastructure, and inhibition of ER-to-Golgi traffic by reducing the levels of these proteins in the cell, interfering with their function with dominant negative forms of the proteins, or by producing non-interacting altered viruses. 2. Determine the role of the rhinovirus IRES in viral host range. We have found that the IRES of HRV2 does not mediate translation of a reporter protein in most organs of adult mice, including the lung. However, HRV2 IRES-mediated translation takes place in many organs of neonatal mice. Experiments in this specific aim are designed to determine whether the IRES of RV16, RV39, and RV1A can function in cells of the mouse lung. These cell cultures will be used to determine if specific structured regions of the IRES prevent internal initiation in primary lung cells from adult mice. Experiments are also planned to determine whether the lack of HRV2 IRES-mediated activity in cultured mouse cells is due to an inhibitor or absence of a required protein, and experiments to identify such factors are proposed. 3. Determine the role of pattern recognition molecules in rhinovirus replication. Experiments in this specific aim are designed to determine the role of specific pattern recognition molecules in rhinovirus replication. We will determine the roles of RIG-I, MDA-5, TLR3, and TLR7 in sensing rhinovirus replication in cultured cells. We have found that RIG-I and MDA-5 are cleaved in cells infected with rhinoviruses. Experiments are planned to determine whether cleavage of these sensor molecules benefits viral replication. In addition, we will determine whether rhinoviruses can replicate in dendritic cells, and whether TLR3 or TLR7 plays a role in sensing the viral genome in these cells. The identification of cell proteins that play a role in rhinovirus replication may reveal new targets for therapeutic intervention of the common cold.

描述（由申请人提供）：人类鼻病毒（HRV）是普通感冒的主要病原体，普通感冒是最常见的人类感染之一。它们也是与哮喘恶化相关的最常见病原体。尽管鼻病毒在医学上具有重要意义，但由于没有方便的动物模型，人们对鼻病毒感染的发病机制知之甚少。该提案中的实验解决了有关鼻病毒生物学的广泛实验问题，包括细胞培养中的复制、宿主范围的分子基础、对感染的先天反应以及在小鼠中建立复制。这些目标将通过以下四个具体目标来实现。 1. 确定与病毒 2B 和 3A 蛋白相互作用的细胞蛋白在鼻病毒复制中的作用。我们已经证明 HRV39 的 3A 蛋白与 GCP60 和 FinGERS（参与囊泡运输的蛋白质）以及参与脂质生物合成的酶 CDIPT（磷脂酰肌醇合酶）相互作用。与 2B 相互作用的细胞蛋白也将被鉴定。我们将通过降低细胞中这些蛋白质的水平，以蛋白质的显性失活形式干扰其功能，来评估这些细胞蛋白质在病毒复制、膜超微结构改变和抑制内质网到高尔基体运输中的作用，或通过产生非相互作用的变异病毒。 2.确定鼻病毒IRES在病毒宿主范围内的作用。我们发现，HRV2 的 IRES 并不介导成年小鼠大多数器官（包括肺）报告蛋白的翻译。然而，HRV2 IRES 介导的翻译发生在新生小鼠的许多器官中。此特定目标的实验旨在确定 RV16、RV39 和 RV1A 的 IRES 是否可以在小鼠肺细胞中发挥作用。这些细胞培养物将用于确定 IRES 的特定结构区域是否阻止成年小鼠原代肺细胞的内部起始。还计划进行实验以确定培养的小鼠细胞中 HRV2 IRES 介导活性的缺乏是否是由于抑制剂或所需蛋白质的缺乏造成的，并提出了鉴定此类因素的实验。 3.确定模式识别分子在鼻病毒复制中的作用。这一特定目标的实验旨在确定特定模式识别分子在鼻病毒复制中的作用。我们将确定 RIG-I、MDA-5、TLR3 和 TLR7 在感测培养细胞中鼻病毒复制中的作用。我们发现 RIG-I 和 MDA-5 在感染鼻病毒的细胞中被切割。计划进行实验以确定这些传感器分子的裂解是否有利于病毒复制。此外，我们还将确定鼻病毒是否可以在树突状细胞中复制，以及TLR3或TLR7是否在感知这些细胞中的病毒基因组中发挥作用。鉴定在鼻病毒复制中发挥作用的细胞蛋白可能会揭示普通感冒治疗干预的新靶点。