Hepatitis C virus host interactions in micropatterened hepatocyte co-cultures

丙型肝炎病毒宿主在微图案化肝细胞共培养物中的相互作用

• 批准号: 7916786
• 负责人: Timothy Patrick Sheahan
• 金额: $ 5.05万
• 依托单位: ROCKEFELLER UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-01 至 2012-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7916786
• 关键词: Acute; Acute Hepatitis C; Affect; Albumins; Antiviral Agents; Antiviral Therapy; Apoptosis; Biological Models; Biomedical Engineering; Cell Line; Cell physiology; Cells; Chronic; Cirrhosis; Coculture Techniques; Data; Development; Disease Outcome; Endothelial Cells; Environment; Family; Fibroblasts; Fibrosis; Flaviviridae; Genes; Genomics; Genotype; Goals; HCV Vaccine; Hematopoietic; Hepatic Stellate Cell; Hepatitis B; Hepatitis C; Hepatitis C virus; Hepatitis C-Like Viruses; Hepatocyte; Human; Immune; Immune response; In Vitro; Infection; Infectious Agent; Inflammatory Response; Integration Host Factors; Lead; Learning; Life Cycle Stages; Liver; Liver Failure; Liver Fibrosis; Lymphocyte; Measures; Messenger RNA; Modeling; Normal Cell; Pathway interactions; Patients; Phenotype; Population; Primary carcinoma of the liver cells; Production; Proteins; Proteomics; Public Health; RNA; RNA Viruses; Recruitment Activity; Research; Role; Staging; Supporting Cell; System; Techniques; Therapeutic; Urea; Vaccines; Virus; Virus Replication; cell type; chemokine; cholangiocyte; cytokine; design; hepatoma cell; in vitro Model; in vivo; in vivo Model; insight; laser capture microdissection; liver transplantation; novel; protective effect; protein expression; research study; tool; virus host interaction; virus pathogenesis

DESCRIPTION (provided by applicant): The hepatitis C virus (HCV) is a major worldwide public health problem affecting an estimated 120-170 million people. Unfortunately, an effective vaccine for HCV does not yet exist and current therapies are toxic and variably effective. While much has been learned about HCV pathogenesis, the virus and host factors that influence spontaneous clearance or persistent infection remain unknown. The lack of robust in vitro and in vivo models that faithfully mimic natural HCV infection of humans continues to hamper studies of HCV pathogenesis. Though hepatocellular carcinoma derived cell lines and immortalized hepatocytes allow us to study the replication of HCV, they are phenotypically and functionally abnormal as compared to primary hepatocytes. While primary hepatocyte cultures are thought to be an ideal model to study HCV infection and virus/host interactions, their liver specific functions rapidly decline in conventional culture systems. Recently, the creation of bioengineered micropatterned co-cultures (MPCCs) of hepatocytes and fibroblasts have allowed for long term culture of hepatocytes with the retention of normal cell function (albumin production, urea synthesis etc). The long-term goal of our research is to gain a better understanding of HCV pathogenesis in order to design specific and effective therapeutics. We hypothesize that early virus and host interactions influence the outcome of disease setting the stage for spontaneous clearance or chronic infection/liver fibrosis. To gain insight into early host response to acute HCV infection in the liver, we propose to characterize HCV virus/host interactions within primary human hepatocyte MPCCs and in MPCCs of increasing cellular complexity. Under specific aim 1, we will differentially phenotype infected and non-infected cells within MPCCs performing both genomic and proteomic profiling to identify host genes and pathways modulated in HCV infection. In specific aim 2, we will increase the cellular complexity of MPCCs to include non-parenchymal cells in order to assess if supporting cell types in the liver contribute to HCV pathogenesis and/or the development of a profibrotic state. In specific aim 3, we will increase the cellular complexity of MPCCs to include lymphocytes resident or recruited to the liver during HCV infection in order to assess the effects of immune cells on acute HCV infection. Through the use of the novel MPCCs, we may gain a better understanding of HCV pathogenesis, which may guide the development of rationally designed antiviral therapies. The development of effective HCV antivirals or vaccines would have a profound impact on global public health.

描述（由申请人提供）：丙型肝炎病毒 (HCV) 是一个主要的全球公共卫生问题，影响估计 120-170 百万人。不幸的是，目前尚不存在有效的丙型肝炎疫苗，并且目前的疗法具有毒性且效果参差不齐。虽然人们对 HCV 发病机制了解很多，但影响自发清除或持续感染的病毒和宿主因素仍然未知。缺乏忠实模拟人类自然 HCV 感染的稳健体外和体内模型继续阻碍 HCV 发病机制的研究。虽然肝细胞癌衍生的细胞系和永生化肝细胞使我们能够研究 HCV 的复制，但与原代肝细胞相比，它们在表型和功能上都存在异常。虽然原代肝细胞培养物被认为是研究 HCV 感染和病毒/宿主相互作用的理想模型，但它们的肝脏特异性功能在传统培养系统中迅速下降。最近，肝细胞和成纤维细胞的生物工程微图案共培养物（MPCC）的创建使得肝细胞可以长期培养，同时保留正常细胞功能（白蛋白产生、尿素合成等）。我们研究的长期目标是更好地了解丙型肝炎病毒的发病机制，以便设计特异性和有效的治疗方法。我们假设早期病毒和宿主的相互作用会影响疾病的结果，为自发清除或慢性感染/肝纤维化奠定基础。为了深入了解宿主对肝脏急性 HCV 感染的早期反应，我们建议表征原代人肝细胞 MPCC 和细胞复杂性增加的 MPCC 中 HCV 病毒/宿主的相互作用。在具体目标 1 下，我们将通过基因组和蛋白质组分析来区分 MPCC 内感染和未感染细胞的表型，以确定 HCV 感染中调节的宿主基因和途径。在具体目标 2 中，我们将增加 MPCC 的细胞复杂性以包括非实质细胞，以评估肝脏中的支持细胞类型是否有助于 HCV 发病机制和/或促纤维化状态的发展。在具体目标 3 中，我们将增加 MPCC 的细胞复杂性，以包括 HCV 感染期间驻留或招募到肝脏的淋巴细胞，以评估免疫细胞对急性 HCV 感染的影响。通过使用新型 MPCC，我们可以更好地了解 HCV 发病机制，从而指导合理设计的抗病毒疗法的开发。开发有效的丙肝病毒抗病毒药物或疫苗将对全球公共卫生产生深远影响。