Murine Model of HCV-Associated Human Liver Cancer

HCV 相关人类肝癌小鼠模型

• 批准号: 8464678
• 负责人: Stanley M. Lemon
• 金额: $ 44.25万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-05-01 至 2017-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8464678
• 关键词: Adopted; Advanced Development; Animal Model; Binding; CD34 gene; Carcinogenesis Mechanism; Cell Culture Techniques; Cells; Chimerism; Chronic Hepatitis C; Complementary DNA; Complex; Country; Coupled; Coupling; Defective Viruses; Development; Dominant-Negative Mutation; Environment; Evaluation; Frequencies; Genes; Genotype; Goals; Hematopoietic stem cells; Hepatic; Hepatitis C; Hepatitis C virus; Hepatocarcinogenesis; Hepatocyte; Human; Human Development; Immune; Immune response; Immune system; Immunocompetent; Incidence; Infection; Inflammation; Lead; Lentivirus Vector; Liver; Liver Fibrosis; Malignant - descriptor; Malignant Neoplasms; Malignant neoplasm of liver; Mediating; Modeling; Molecular; Molecular Genetics; Morbidity - disease rate; Mus; Outcome; Oxidative Stress; Pathogenicity; Patients; Play; Primary carcinoma of the liver cells; Protocols documentation; Publishing; Recombinant DNA; Recombinants; Retinoblastoma; Retinoblastoma Protein; Risk; Role; SV40 T Antigens; Stem cells; Subfamily lentivirinae; Support System; System; T cell response; TACSTD2 gene; Testing; Therapeutic; Transgenes; Transplantation; Tumor Suppressor Proteins; Viral; Viral Pathogenesis; Virus; Virus Replication; Work; c-Myc Staining Method; carcinogenesis; fetal; follow-up; hepatitis C virus nucleocapsid protein; liver inflammation; mortality; mouse model; mutant; novel; oxidative DNA damage; response; sphingosine kinase; tumorigenesis; viral carcinogenesis; virus genetics; virus pathogenesis

DESCRIPTION (provided by applicant): Chronic hepatitis C virus (HCV) infection is now the leading cause of hepatocellular carcinoma (HCC), a highly lethal cancer with rapidly increasing incidence in the U.S. How HCV causes liver cancer is not well understood because of the lack of a small animal model for HCV pathogenesis. The overarching goal of this interdisciplinary, multi-PI project is to refine a recently developed humanized mouse model of hepatitis C (the AFC8-hu mouse) that possesses both a chimeric human liver and a functioning human immune system, and to use this model to elucidate viral mechanisms of carcinogenesis responsible for HCV-associated HCC. AFC8- hu mice engrafted with CD34+ human hematopoietic stem cells and EpCAM+ human hepatoblasts and hepatic stem cells achieve ~18% hepatic chimerism, are permissive for HCV infection, and develop a human HCV- specific T cell response with hepatic inflammation when infected with HCV. Remarkably, HCV infection of these mice results in significant hepatic fibrosis and evidence of oxidative DNA damage - hallmarks of chronic hepatitis C in human patients. Three interrelated specific aims are proposed to refine this model and to apply it to the study of the mechanisms of carcinogenesis responsible for HCV-associated HCC. Aim 1 focuses on optimizing protocols to enhance hepatic chimerism in AFC8-hu mice, and to infect AFC8-hu mice with genotype 1a HCVcc produced in cell culture from recombinant DNA, thereby providing a system supporting reverse molecular viral genetics studies of HCV-mediated carcinogenesis. In Aim 2, p53 will be inactivated in human hepatoblasts prior to transplantation into AFC8 mice to provide for accelerated development of human liver cancer in HCV-infected mice, and to facilitate discovery of novel determinants of HCV-mediated hepatocarcinogenesis. Aim 3 will utilize a reverse molecular genetics strategy to test the hypothesis that the disruption of critical tumor suppressor function by HCV promotes cancer in the genotoxic environment of immune-mediated hepatic inflammation and oxidative stress, and to specifically assess the role played by the targeted destruction of the retinoblastoma protein by HCV and sequestration of the tumor suppressor DDX3 by HCV core protein. Further development of the AFC8-hu mouse model, coupled with the use of infectious genotype 1a H77S virus produced from recombinant cDNA, will provide a robust and completely unique experimental system in which the complex interplay of direct and indirect mechanisms of carcinogenesis can be elucidated. The results of these studies will add significantly to current understanding of how HCV infection leads to the development of HCC, and advance the development and evaluation of new preventative and therapeutic strategies to lessen the considerable burden of morbidity and mortality imposed by liver cancer in patients with chronic hepatitis C.

描述（由申请人提供）：慢性丙型肝炎病毒（HCV）感染现在是肝细胞癌（HCC）的主要原因，这是一种高度致死性的癌症，在美国，HCV在美国的发病率迅速增加，因为缺乏HCV病原体的小型动物模型，因此肝癌的发病率不佳。这个跨学科的多PI型项目的总体目标是完善具有嵌合具有嵌合人类肝脏和功能正常的人类免疫系统的最近开发的人性化小鼠模型（AFC8-HU小鼠），并使用此模型来阐明负责HCV与HCV与HCCAPIADIAD HCC的癌症的病毒机制。植入CD34+人造血干细胞和Epcam+人肝细胞的AFC8-HU小鼠，肝干细胞的AFC8小鼠可实现〜18％的肝嵌合感，允许HCV感染，并在HCV感染HCV时会产生人类HCV-特异性T细胞炎症。值得注意的是，这些小鼠的HCV感染会导致肝纤维化显着，并表现出氧化性DNA损伤的证据 - 人类患者慢性丙型肝炎的标志。提出了三个相互关联的特定目的来完善该模型，并将其应用于负责HCV相关HCC的致癌机制的研究。 AIM 1专注于优化方案，以增强AFC8-HU小鼠的肝嵌合体，并用重组DNA在细胞培养中产生的基因型1A HCVCC感染AFC8-HU小鼠，从而提供了一个支持HCV介导的介导的营养性营养的系统，从而提供了一个支持反向分子病毒遗传学研究的系统。在AIM 2中，在人肝细胞中将p53灭活，然后将其移植到AFC8小鼠中，以提供HCV感染的小鼠中人肝癌的加速发展，并促进发现HCV介导的肝癌的新型决定因素。 Aim 3 will utilize a reverse molecular genetics strategy to test the hypothesis that the disruption of critical tumor suppressor function by HCV promotes cancer in the genotoxic environment of immune-mediated hepatic inflammation and oxidative stress, and to specifically assess the role played by the targeted destruction of the retinoblastoma protein by HCV and sequestration of the tumor suppressor DDX3 by HCV core protein. AFC8-HU小鼠模型的进一步开发，再加上由重组cDNA产生的感染性基因型1A H77S病毒，将提供一个可靠且完全独特的实验系统，在该系统中，可以阐明直接和间接机制的复杂相互作用。这些研究的结果将大大增加对HCV感染如何导致HCC发展的最新理解，并提高对新的预防和治疗策略的发展和评估，以减轻慢性肝炎患者肝癌施加的大量发病率和死亡率负担。