Function of Lipid Droplets in Viral Entry and Membrane Fusion

脂滴在病毒进入和膜融合中的功能

基本信息

批准号：
8679468
负责人：
HENGLI TANG
金额：
$ 22.03万
依托单位：
FLORIDA STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-04-01 至 2016-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8679468
关键词：
Affinity Antiviral Agents Biochemical Capsid Cell Communication Cell Death Cell Surface Receptors Cells Cellular Structures Chemicals Clathrin Complex Cytoplasm Data Dengue Virus Development Diagnostic Drug Targeting Drug resistance Early Endosome Endocytosis Endosomes Face Fractionation Genome Goals Health Hepatitis C Hepatitis C virus Human Human Cell Line Infection Intervention Knock-out Knowledge Life Cycle Stages Lipids Mediating Membrane Fusion Modeling Organelles Patients Pharmaceutical Preparations Process Proteins RNA RNA Interference RNA Viruses Recruitment Activity Reporting Research Role System Technology Vaccines Viral Virion Virus Virus Diseases Work base cofactor genetic manipulation inhibitor/antagonist insight knockout gene new therapeutic target novel nuclease particle pathogen programs prophylactic public health relevance virus host interaction

Affinity Antiviral Agents Biochemical Capsid Cell Communication Cell Death Cell Surface Receptors Cells Cellular Structures Chemicals Clathrin Complex Cytoplasm Data Dengue Virus Development Diagnostic Drug Targeting Drug resistance Early Endosome Endocytosis Endosomes Face Fractionation Genome Goals Health Hepatitis C Hepatitis C virus Human Human Cell Line Infection Intervention Knock-out Knowledge Life Cycle Stages Lipids Mediating Membrane Fusion Modeling Organelles Patients Pharmaceutical Preparations Process Proteins RNA RNA Interference RNA Viruses Recruitment Activity Reporting Research Role System Technology Vaccines Viral Virion Virus Virus Diseases Work base cofactor genetic manipulation inhibitor/antagonist insight knockout gene new therapeutic target novel nuclease particle pathogen programs prophylactic public health relevance virus host interaction

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项目摘要

DESCRIPTION (provided by applicant): The broad, long-term goal of our research program is to advance knowledge of virus-host cell interactions by characterizing cellular cofactors essential for viral infections, particularly positive-strand RNA viruses such as hepatitis C virus (HCV) and dengue virus (DENV). These viruses infect millions of people worldwide and pose major threats to human health. Understanding how these viruses interact with the host cell is of critical importance to the development of diagnostics, antiviral drugs, and a prophylactic vaccine. In preliminary studies, we have uncovered a critical role of lipid droplets and one of its associated proteins in DENV and HCV entry into host cells at the membrane fusion step. Although a role of the lipid droplets in viral assembly has been reported, a function of these organelles in viral entry and membrane fusion has not been proposed before. In this study, we will investigate the mechanism of action for lipid droplets and its associated protein, CIDEB, to facilitate the entry of diverse RNA viruses. The following specific aims are proposed: (1) to define the roles of lipid droplets and their recruitment of early endosomes in virus entry. Our working hypothesis of this aim is that LDs and the recruitment of virus-containing early endosomes to LDs are required for completing viral entry. We will use pharmacological and genetic manipulations to reduce LDs in the target cells of DENV and HCV and then determine if infection by these viruses is inhibited at specific steps such as endocytosis, membrane fusion, or capsid release. (2) to determine the function of CIDEB during the infection cycle of positive-strand RNA viruses. We have demonstrated the CIDEB knockdown (KD) with RNAi reduced DENV/HCV infection and hypothesize that CIDEB can serve as a new host target for treating infections by positive-stranded RNA viruses. We will use CIDEB knockout (KO) models, generated using TAL effector nuclease (TALEN) technology, to determine the importance of CIDEB in the life cycles of various RNA viruses. The results of the proposed studies will not only provide significant insights into the virus- and host-related functions of lipid droplets and CIDEB but also may reveal new therapeutic targets for antiviral intervention directed at medically important RNA viruses.

描述（由申请人提供）：我们的研究计划的广泛长期目标是通过表征对病毒感染所必需的细胞辅助因子，尤其是阳性链链RNA病毒（例如乙型肝炎病毒（HCV）和登革热病毒（DENV）所必需的细胞辅助因子（尤其是阳性链链RNA病毒）的知识。这些病毒在全球范围内感染了数百万的人，并对人类健康构成了重大威胁。了解这些病毒如何与宿主细胞相互作用对于诊断，抗病毒药物和预防性疫苗的发展至关重要。在初步研究中，我们发现了脂质液滴及其其中一种的关键作用在膜融合步骤中，DENV和HCV进入宿主细胞中的相关蛋白质。尽管已经报道了脂质液滴在病毒组装中的作用，但以前尚未提出这些细胞器在病毒进入和膜融合中的功能。在这项研究中，我们将研究脂质液滴及其相关蛋白CIDEB的作用机理，以促进各种RNA病毒的进入。提出了以下特定目的：（1）定义脂质液滴的作用及其在病毒进入中的早期内体的募集。我们对此目标的工作假设是，LDS和募集含病毒的早期内体向LDS募集是完成病毒入口所必需的。我们将使用药理学和遗传操作来减少DENV和HCV靶细胞中的LDS，然后确定这些病毒感染是否在特定步骤（例如内吞作用，膜融合或衣壳释放）下抑制。（2）确定在阳性链RNA病毒的感染周期中CIDEB的功能。我们已经证明了Cideb敲低（KD）的RNAi降低了DENV/HCV感染，并假设CideB可以作为治疗阳性链RNA病毒感染的新宿主靶标。我们将使用使用TAL效应核酸酶（TALEN）技术生成的CIDEB敲除（KO）模型来确定CIDEB在各种RNA病毒的生命周期中的重要性。拟议的研究的结果不仅将提供有关脂质液滴和CIDEB病毒和宿主相关功能的重大见解，而且还可能揭示了针对医学上重要的RNA病毒的抗病毒干预的新的治疗靶标。