Role of the inflammasome in hepatitis C virus pathogenesis

炎症小体在丙型肝炎病毒发病机制中的作用

• 批准号: 8507828
• 负责人: GULAM WARIS
• 金额: $ 38.63万
• 依托单位: ROSALIND FRANKLIN UNIV OF MEDICINE & SCI
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-08-01 至 2014-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8507828
• 关键词: Anticholesteremic Agents; Area; Attention; Binding Proteins; Biological Assay; Caspase-1; Cells; Cholesterol; Chronic; Chronic Hepatitis C; Cleaved cell; Complex; Confocal Microscopy; Data; Dinoprostone; Employee Strikes; Environment; Equipment; Fatty Acids; Fatty Liver; Fluorescence; Genotype; Goals; Golgi Apparatus; Hepatitis C; Hepatitis C virus; Hepatocyte; Homeostasis; Human; Immune; Infection; Inflammation; Inflammatory; Insulin Resistance; Knowledge; Laboratories; Life Cycle Stages; Lipids; Liver; Liver Fibrosis; Liver diseases; Malignant Neoplasms; Mass Spectrum Analysis; Mediating; Metabolic Pathway; Methods; Mission; Molecular; Morphogenesis; Obesity; Pathogenesis; Pathway interactions; Patients; Play; Primary carcinoma of the liver cells; Process; Production; Protein Precursors; Proteins; Proteomics; Public Health; Regulation; Regulatory Element; Reporter; Research; Research Personnel; Resources; Reverse Transcriptase Polymerase Chain Reaction; Risk Factors; Role; Signal Pathway; Small Interfering RNA; Staging; Sterols; Technology; Testing; Time; Toxin; United States National Institutes of Health; Very low density lipoprotein; Viral; Viral Proteins; Virus; Virus Replication; Western Blotting; Work; base; design; fatty acid metabolism; innovation; insight; lipid metabolism; multidisciplinary; novel; novel strategies; protein activation; protein protein interaction; response; tumorigenesis; virus pathogenesis

DESCRIPTION (provided by applicant): The role of host lipid metabolism has attracted considerable attention in hepatitis C virus (HCV) life cycle. Altered host lipid metabolism is the cause of fatty-liver disease. There is a fundamental gap in understanding how HCV infection induces the proteolytic activation of sterol regulatory element binding proteins (SREBPs), the master regulator of fatty-liver disease, which is observed in patients with chronic HCV infection. The long- term goal of this research is to define the molecular mechanisms of altered lipid homeostasis that will better characterize the pathogenesis of HCV-mediated liver disease. The objective of this proposal is to identify the molecular mechanisms of activation of SREBPs by HCV. The central hypothesis of the application is that induction of the caspase-1 inflammasome complex in HCV-infected cells mediates the activation of SREBPs. The rationale for the proposed research is that, by understanding how HCV-mediated caspase-1 inflammasome induces the activation of SREBPs, their activation can be inhibited in new and innovative approaches to arrest the production of HCV and the progression of liver disease associated with HCV-infection. Thus, the proposed research is relevant to NIH mission that pertains to developing fundamental knowledge that will potentially help design the alternate strategies in the treatment of chronic hepatitis C infection. Supported by strong preliminary data, the proposal's main hypothesis will be tested under two specific aims: 1) Determine the mechanism of caspase-1 activation by HCV; 2) Define the mechanism of SREBPs proteolytic activation. Under the first aim, the role of liver-specific inflammasome complex in caspase-1 activation will be investigated using novel bimolecular fluorescence complementation (BiFC) assay, proteomics/mass spectroscopy, real-time RT-PCR, siRNA technology, confocal microscopy, and protein-protein interaction. Under the second aim, the role of the components of the inflammasome complex such as, NALP3, ASC, and caspase-1 on the regulation of SCAP, Insig, and COPII proteins will be investigated using the similar methods as described for aim 1. The activation of SREBPs will be investigated using western-blot, confocal microscopy, and cell-based reporter assays. The results from these studies will yield novel insights into mechanisms of altered lipid homeostasis, liver oncogenesis, insulin resistance, and obesity associated with chronic HCV. At the completion of these studies our work is collectively expected to establish the potential role of HCV in the activation of caspase-1 inflammasome complex and proteolytic activation of SREBPs through the interactions with chronic inflammatory processes. This work will also define the novel caspase-1 inflammasome complex induction in human hepatocytes. The proposed research is significant because it is expected to provide novel strategies for targeting the viral or cellular determinants to arrest the production of HCV as well as the progression of liver disease associated with HCV-infection. In addition, it is expected that the results will fundamentally advance the fields of liver inflammation and lipid metabolism mediated by HCV-infection.

描述（由申请人提供）：宿主脂质代谢的作用引起了丙型肝炎病毒（HCV）生命周期的极大关注。宿主脂质代谢改变是脂肪肝病的原因。了解HCV感染如何诱导固醇调节元素结合蛋白（SREBPS）的蛋白水解激活（SREBPS）是脂肪肝疾病的主要调节剂，这在慢性HCV感染患者中观察到。这项研究的长期目标是定义脂质稳态改变的分子机制，这将更好地表征HCV介导的肝病的发病机理。该建议的目的是确定HCV激活SREBP的分子机制。该应用的中心假设是，HCV感染细胞中caspase-1炎症体复合物的诱导介导了SREBP的激活。拟议研究的基本原理是，通过了解HCV介导的caspase-1炎性体如何诱导SREBP的激活，可以在新颖的创新方法中抑制它们的激活，以阻止HCV的产生以及与HCV感染相关的肝脏疾病的进展。因此，拟议的研究与NIH的使命相关，该任务与开发基本知识有关，这些知识将有可能帮助设计慢性丙型肝炎感染的替代策略。 在强大的初步数据的支持下，该提案的主要假设将在两个具体目的下进行检验：1）确定HCV通过HCV激活Caspase-1的机理； 2）定义SREBPS蛋白水解活化的机制。在第一个目标下，将使用新型的双分子荧光互补（BIFC）测定，蛋白质组学/质谱，实时RT-PCR，SIRNA技术，共焦显微镜和蛋白质 - 蛋白质蛋白质相互作用来研究肝脏特异性炎症体复合物在caspase-1激活中的作用。在第二个目标下，将使用与AIM 1所述的类似方法一起研究炎性体复合物的成分（例如，NALP3，ASC和CASPASE-1）对SCAP，INMIG和COPII蛋白的调节的作用。将使用SREBPS的激活进行使用，使用SREBPS的激活，使用Western-blot，Contest-blot，Condocal Microscopy，Coldocal Microscopy，Cell Ectial Indues Cell Indust Imases Reporter和Cell Indues Reporter sass。这些研究的结果将产生对改变脂质稳态，肝癌发生，胰岛素抵抗和与慢性HCV相关的肥胖的机制的新见解。这些研究完成后，我们的工作被共同期望，可以通过与慢性炎症过程的相互作用来确定HCV在Caspase-1炎症体复合物激活中的潜在作用和SREBP的蛋白水解激活。这项工作还将定义人肝细胞中新型的caspase-1炎症体复合诱导。拟议的研究很重要，因为预计将提供靶向病毒或细胞决定因素以阻止HCV产生以及与HCV感染相关的肝病的进展的新型策略。此外，预计结果将从根本上推进HCV感染介导的肝脏炎症和脂质代谢的田间。

项目成果

Role of hepatitis C virus induced osteopontin in epithelial to mesenchymal transition, migration and invasion of hepatocytes.

• DOI: 10.1371/journal.pone.0087464
• 发表时间: 2014
• 期刊: PloS one
• 影响因子: 3.7
• 作者: Iqbal J;McRae S;Mai T;Banaudha K;Sarkar-Dutta M;Waris G
• 通讯作者: Waris G

The Hepatitis C Virus-induced NLRP3 Inflammasome Activates the Sterol Regulatory Element-binding Protein (SREBP) and Regulates Lipid Metabolism.

丙型肝炎病毒诱导的 NLRP3 炎症小体激活甾醇调节元件结合蛋白 (SREBP) 并调节脂质代谢。

• DOI: 10.1074/jbc.m115.694059
• 发表时间: 2016
• 期刊: The Journal of biological chemistry
• 作者: McRae,Steven;Iqbal,Jawed;Sarkar-Dutta,Mehuli;Lane,Samantha;Nagaraj,Abhiram;Ali,Naushad;Waris,Gulam
• 通讯作者: Waris,Gulam