Hepatitis C virus polarized cell entry pathways

丙型肝炎病毒极化细胞进入途径

基本信息

批准号：
8396182
负责人：
Matthew J Evans
金额：
$ 36.53万
依托单位：
ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-08-15 至 2017-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8396182
关键词：
Address Adverse effects Affect Animal Model Antiviral Agents Biochemical Genetics Biological Assay Biological Models CD81 gene Cell Culture Techniques Cell Line Cell Polarity Cell physiology Cells Clathrin Complement Derivation procedure Development Endocytosis Endosomes Event Foundations Genetic Glycoproteins Glycosaminoglycans Goals Health Hepatitis C Hepatitis C virus Hepatocyte Human Individual Infection Integration Host Factors Kinetics Knowledge Label Life Life Cycle Stages Light Liver Location Low Density Lipoprotein Receptor Malignant Neoplasms Malignant neoplasm of liver Mediating Methods MicroRNAs Modeling Mus Mutation Nature Pan Genus Pathway interactions Peptide antibodies Process Proteins Public Health RNA replication Relative (related person)Research Resistance Resistance development Risk Role SR-BI receptor Staging System Testing Tight Junctions Time Viral Viral Proteins Virion Virus Virus Inhibitors Virus Replication Visual base claudin-1 protein genetic analysis human PHEMX protein in vivo inhibitor/antagonist innovation insight mutant novel occludin polarized cell receptor research study viral RNA

项目摘要

DESCRIPTION (provided by applicant): The hepatitis C virus (HCV) is the leading cause of liver cancer in the Western Hemisphere. Over 170 million people are infected with this virus and thus at increased risk of liver damage and cancer. HCV infection is curable, and thus such health consequences are completely avoidable. However the current HCV therapy is inadequate, as it is associated with severe side effects and only effective in about half of treate individuals. Effective HCV treatment will require antivirals targeting multiple HCV replication steps that can be used in combination to avoid the derivation of resistant viruses. The HCV cell entry process is an attractive target for antiviral development. Inhibiting this stage of the virallife cycle would curtail the persistence of HCV in vivo, which requires constant infection of new cells, as well as the spread of viral mutants that have developed resistance to other HCV inhibitors. The research proposed in this application will advance our understanding of HCV cell entry mechanisms, with the ultimate goal of uncovering novel antiviral targets. Although we now know that numerous host factors are required for HCV cell entry, we do not understand how they mediate HCV infection. The experiments described in this application are based on the hypothesis that the incoming HCV virion utilizes each of these cellular factors in a sequential manner to mediate cell entry. As two of the host proteins that are required for HCV cell entry are tight junction proteins, it is likely that cell polarity, which regulates the localization and funcions of these proteins, impacts the HCV cell entry process. Although the majority of HCV cell entry studies have been conducted in non-polarized cells, we have recently developed the first polarized cell system that permits efficient infection of authentic HCV grown in cell culture. This system will serve as the foundation for the studies proposed in this application. Specifically, we propose experiments to compare HCV cell entry pathways in non-polarized and polarized using a variety of HCV cell entry factor specific inhibitors. Furthermore, we will test how modulating cell polarity influences HCV cell entry, and examine the host factor requirements with entry factor specific inhibitors, silencing, and complementation with a variety of mutants. We will also analyze when and where each entry factor is used by analyzing the kinetics of action of entry factor specific inhibitors and visualizing the HCV cell entry process in non-polarized and polarized cells. Finally, we will identify physical and genetic interactions between host factors and the incoming HCV virion, and explore how such interactions are affected by cell polarity. PUBLIC HEALTH RELEVANCE: The development of novel, more effective, and better-tolerated therapies targeting the hepatitis C virus (HCV) has been hampered by an incomplete understanding of its basic mechanisms of replication. The experiments in this proposal are aimed at providing an in-depth understanding of how the HCV virion enters a host cell with a focus on how cell polarity regulates this process and how cellular proteins we recently found to be essential for this process, interact with and are utilized by the incoming virion. Such information will be directly relevant to the development of HCV antivirals and small animal models.

描述（由申请人提供）：丙型肝炎病毒（HCV）是西半球肝癌的主要原因。超过1.7亿人感染了这种病毒，因此有增加肝脏损害和癌症的风险。 HCV感染是可以治愈的，因此这种健康后果是完全可以避免的。但是，目前的HCV疗法是不充分的，因为它与严重的副作用有关，并且仅在大约一半的治疗个体中有效。有效的HCV治疗将需要针对多个HCV复制步骤的抗病毒药，这些步骤可用于避免抗性病毒的推导。 HCV细胞进入过程是抗病毒发育的有吸引力的目标。抑制Virallife周期的这一阶段将减少体内HCV的持久性，这需要持续感染新细胞，以及对其他HCV抑制剂产生抗性的病毒突变体的扩散。本应用程序中提出的研究将提高我们对HCV细胞进入机制的理解，其最终目标是发现新型的抗病毒靶标。尽管我们现在知道HCV细胞进入需要许多宿主因素，但我们不了解它们如何介导HCV感染。本应用中描述的实验基于以下假设：传入的HCV病毒体以顺序的方式利用这些细胞因子中的每一个来介导细胞进入。由于HCV细胞进入所需的两个宿主蛋白是紧密的连接蛋白，因此调节这些蛋白质的定位和功能的细胞极性可能会影响HCV细胞进入过程。尽管大多数HCV细胞进入研究是在非极化细胞中进行的，但我们最近开发了第一个极化细胞系统，该系统允许在细胞培养中有效感染地正质HCV。这系统将作为本应用程序提出的研究的基础。具体而言，我们提出了实验，以比较使用多种HCV细胞进入因子特异性抑制剂中非极化和极化的HCV细胞进入途径。此外，我们将测试调节细胞极性如何影响HCV细胞的进入，并使用特定于入口因子特异性抑制剂，沉默和与多种突变体的补充检查宿主因子需求。我们还将通过分析特定于入口因子特异性抑制剂的动作动力学并在非极化和极化细胞中可视化HCV细胞进入过程，从而分析何时何地使用每个入口因子。最后，我们将确定宿主因子与传入的HCV病毒体之间的物理和遗传相互作用，并探讨这种相互作用如何受细胞极性影响。公共卫生相关性：针对丙型肝炎病毒（HCV）的新颖，更有效和耐受性较高的疗法的发展受到了对其基本复制机制的不完全理解。该提案中的实验旨在深入了解HCV病毒体如何进入宿主细胞，重点关注细胞极性如何调节这一过程，以及我们最近发现的细胞蛋白如何在此过程中与该过程相互作用并被收入的Virion使用并使用。此类信息将与HCV抗病毒药和小动物模型的发展直接相关。