Hepatitis C Virus interaction with the innate immune system in HepG2-HFL cells

丙型肝炎病毒与 HepG2-HFL 细胞中先天免疫系统的相互作用

基本信息

批准号：
8703101
负责人：
Benjamin Goldman-Israelow
金额：
$ 4.77万
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：
2012-06-01 至 2015-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8703101
关键词：
Acute Liver Failure Adverse effects Affect Animal Model Antiviral Agents Antiviral Response Biological Models CD81 gene Cell Communication Cell Culture Techniques Cell Line Cells Chronic Chronic Hepatitis C Cirrhosis Cleaved cell Development Disease Employee Strikes Exhibits Genes Goals HCV Vaccine Hepatitis C Hepatitis C virus Hepatocyte Human Immune Immune response Immune system Impairment Infection Inflammation Inflammatory Interferons Life Life Cycle Stages Link Liver Liver diseases Malignant neoplasm of liver MicroRNAs Mitochondria Modeling Molecular Molecular Biology Techniques Nature Outcome Output Pathogenesis Pathway interactions Patients Peptide Hydrolases Play Population Primary carcinoma of the liver cells Process Production RNA Helicase Reaction Research Role Side Signal Transduction Solutions Staging Stimulus System Testing Therapeutic Time Tretinoin Viral Viral Proteins Virus Virus Diseases Virus Replication Work adaptive immunity base combat cytokine human TLR3 protein improved in vivo insight liver injury liver transplantation melanoma novel novel therapeutics pathogen prevent public health relevance receptor research study response sensor viral RNA virus host interaction virus pathogenesis

项目摘要

DESCRIPTION (provided by applicant): Hepatitis C virus is the leading cause of liver disease in the Western hemisphere. The current HCV therapy is inadequate, as it is associated with severe side effects and often not effective. The majority of HCV infections are persistent, life-long chronic infections that slowly induce pathogenesis. Furthermore, HCV does not appear to directly induce liver disease, rather it does so indirectly by stimulating cytokine production and inflammation, which suggests that its impairment of innate immune pathways is not absolute. The virus-host interactions and molecular mechanisms that support persistent HCV replication and chronic infection, and those that eventually induce liver disease are not understood. We hypothesize that innate intracellular immune defenses play an essential role in regulating HCV replication and that viral persistence is linked to HCV control of innate defense processes. Nearly all the studies of this topic have been conducted in Huh-7 cells or derivatives, which exhibit severely impaired innate immune responses. We recently showed that hepatoma-derived HepG2 cells efficiently support the entire HCV life cycle if the liver-specific microRNA miR-122 is expressed along with the missing entry factor CD-81 (HepG2-HFL cells). While HepG2 cells containing these two factors can replicate the entire life cycle of HCV, we found that there was a striking difference in these cells' ability to support infection and spread when compared to the previously established Huh-7.5 cell line. At the same time, we found that the innate immune response in these cells was more active in response to HCV infection. Therefore, we hypothesize that HepG2-HFL cells will faithfully recapitulate the innate immune responses present in hepatocytes, and thus provide a better model through which to understand the interactions between HCV and the innate immune system. Our ultimate goal is to better understand how HCV can subvert the antiviral response while allowing for activation of inflammatory pathways. We propose experiments to compare innate immune pathways in HepG2-HFL cells with those of primary human hepatocytes and Huh-7 cells, with a particular focus on the responses induced by the cellular viral RNA sensors RIG-I, MDA5, and TLR3. We will next examine the interaction between HCV and innate immune pathways in HepG2-HFL cells to determine whether HCV activates innate immune responses that can effectively combat infection as well as inhibit such responses through the action of viral proteins. We anticipate tha the results of these studies will provide novel insights into the interaction between HCV and the innate immune system, which remains poorly understood due to a lack of robust model systems, despite its importance in the pathogenesis of associated disease.

描述（由申请人提供）：乙型肝炎病毒是西半球肝病的主要原因。当前的HCV疗法是不充分的，因为它与严重的副作用相关，并且通常无效。大多数HCV感染是持续的，终生的慢性感染，会缓慢诱发发病机理。此外，HCV似乎并未直接诱导肝病，而是通过刺激细胞因子的产生和炎症而间接地诱导肝病，这表明其先天免疫途径的损害不是绝对的。尚不清楚支持持续性HCV复制和慢性感染的病毒宿主相互作用和分子机制，以及最终诱导肝病的病毒相互作用和机制。我们假设先天的细胞内免疫防御在调节HCV复制中起着至关重要的作用，并且病毒持续性与HCV对先天防御过程的控制有关。该主题的几乎所有研究都是在HUH-7细胞或衍生物中进行的，这些细胞或衍生物表现出严重受损的先天免疫反应。我们最近表明，如果肝脏特异性micrORNA miR-122与缺失的入口因子CD-81（HEPG2-HFL细胞）一起表达，则肝癌衍生的HEPG2细胞有效地支持整个HCV生命周期。虽然包含这两个因素的HEPG2细胞可以复制HCV的整个生命周期，但我们发现，与先前确定的HuH-7.5细胞系相比，这些细胞支持感染和扩散的能力存在显着差异。同时，我们发现这些细胞中的先天免疫反应在响应HCV感染的情况下更为活跃。因此，我们假设HEPG2-HFL细胞将忠实地概括肝细胞中存在的先天免疫反应，从而提供了一个更好的模型，可以通过该模型来了解HCV与先天免疫系统之间的相互作用。我们的最终目标是更好地了解HCV如何在允许激活炎症途径的同时颠覆抗病毒反应。我们提出了将HEPG2-HFL细胞中的先天免疫途径与原代人肝细胞和HUH-7细胞的实验进行比较，并特别关注由细胞病毒RNA传感器RIG-I，MDA5和TLR3引起的反应。接下来，我们将研究HEPG2-HFL细胞中HCV与先天免疫途径之间的相互作用，以确定HCV是否激活了先天免疫反应，该反应是否可以有效地对抗感染并通过病毒蛋白的作用来抑制此类反应。我们预计，这些研究的结果将为HCV与先天免疫系统之间的相互作用提供新的见解，尽管缺乏强大的模型系统，尽管它在相关疾病的发病机理中，但由于缺乏强大的模型系统，它们仍然很少理解。