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：
8512570
负责人：
Benjamin Goldman-Israelow
金额：
$ 4.72万
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/8512570
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）：丙型肝炎病毒是西半球肝脏疾病的主要原因。目前的 HCV 疗法还不够充分，因为它会产生严重的副作用，而且往往无效。大多数 HCV 感染是持续性、终生慢性感染，缓慢诱发发病。此外，HCV似乎不会直接诱发肝脏疾病，而是通过刺激细胞因子的产生和炎症来间接诱发肝脏疾病，这表明它对先天免疫途径的损害并不是绝对的。支持丙型肝炎病毒持续复制和慢性感染以及最终诱发肝病的病毒与宿主相互作用和分子机制尚不清楚。我们假设先天细胞内免疫防御在调节 HCV 复制中发挥重要作用，并且病毒持续存在与 HCV 对先天防御过程的控制有关。几乎所有该主题的研究都是在 Huh-7 细胞或其衍生物中进行的，这些细胞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 与缺失的进入因子 CD-81（HepG2-HFL 细胞）一起表达，则肝癌来源的 HepG2 细胞可有效支持整个 HCV 生命周期。虽然含有这两个因子的 HepG2 细胞可以复制 HCV 的整个生命周期，但我们发现与之前建立的 Huh-7.5 细胞系相比，这些细胞支持感染和传播的能力存在显着差异。同时，我们发现这些细胞的先天免疫反应在应对HCV感染时更加活跃。因此，我们假设 HepG2-HFL 细胞将忠实地再现肝细胞中存在的先天免疫反应，从而提供一个更好的模型来了解 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丙型肝炎病毒如何破坏抗病毒反应，同时激活炎症途径。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 细胞中的先天免疫途径进行比较的实验，特别关注细胞病毒 RNA 传感器 RIG-I、MDA5 和 TLR3 诱导的反应。接下来我们将检查 HCV 与 HepG2-HFL 细胞中先天免疫途径之间的相互作用，以确定 HCV 是否激活可有效对抗感染的先天免疫反应，并通过病毒蛋白的作用抑制此类反应。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，尽管它在相关疾病的发病机制中很重要，但由于缺乏可靠的模型系统，人们对这种相互作用仍然知之甚少。