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）：宿主脂质代谢在丙型肝炎病毒（HCV）生命周期中的作用引起了相当多的关注。宿主脂质代谢改变是脂肪肝疾病的原因。在理解 HCV 感染如何诱导甾醇调节元件结合蛋白 (SREBP) 的蛋白水解激活方面存在根本性的差距，SREBP 是脂肪肝疾病的主要调节因子，在慢性 HCV 感染患者中观察到这种情况。这项研究的长期目标是确定脂质稳态改变的分子机制，从而更好地表征 HCV 介导的肝病的发病机制。该提案的目的是确定 HCV 激活 SREBP 的分子机制。该申请的中心假设是 HCV 感染细胞中 caspase-1 炎性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，可以通过新的创新方法抑制 SREBP 的激活，从而阻止 HCV 的产生以及与 HCV 相关的肝病的进展-感染。因此，拟议的研究与 NIH 的使命相关，该使命涉及发展基础知识，这可能有助于设计治疗慢性丙型肝炎感染的替代策略。 在强有力的初步数据支持下，该提案的主要假设将在两个具体目标下得到检验：1）确定HCV激活caspase-1的机制； 2) 定义SREBPs蛋白水解激活机制。第一个目标是使用新型双分子荧光互补 (BiFC) 测定、蛋白质组学/质谱、实时 RT-PCR、siRNA 技术、共聚焦显微镜和技术来研究肝脏特异性炎症体复合物在 caspase-1 激活中的作用。蛋白质-蛋白质相互作用。在第二个目标下，将使用与目标 1 中所述类似的方法研究炎症体复合物的成分（例如 NALP3、ASC 和 caspase-1）对 SCAP、Insig 和 COPII 蛋白调节的作用。将使用蛋白质印迹、共聚焦显微镜和基于细胞的报告基因检测来研究 SREBP 的激活。这些研究的结果将为改变脂质稳态、肝脏肿瘤发生、胰岛素抵抗和与慢性丙型肝炎相关的肥胖机制提供新的见解。这些研究完成后，我们的工作预计将通过与慢性炎症过程的相互作用，确定 HCV 在 caspase-1 炎性体复合物激活和 SREBP 蛋白水解激活中的潜在作用。这项工作还将定义人肝细胞中新型 caspase-1 炎性体复合物的诱导。拟议的研究意义重大，因为它有望提供针对病毒或细胞决定因素的新策略，以阻止 HCV 的产生以及与 HCV 感染相关的肝病的进展。此外，预计该结果将从根本上推进HCV感染介导的肝脏炎症和脂质代谢领域的发展。

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