Hepatic differentiation of stem cells and the cellular determinants of hepatitis

干细胞的肝分化和肝炎的细胞决定因素

• 批准号: 8728449
• 负责人: HENGLI TANG
• 金额: $ 28.88万
• 依托单位: FLORIDA STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-09-12 至 2014-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8728449
• 关键词: Address; Affect; Animal Model; Animals; Antiviral Agents; Atlases; Binding; Bioinformatics; Biological Models; Candidate Disease Gene; Cell Communication; Cell Culture Techniques; Cell Death; Cells; Complementary DNA; Development; Diagnostic; Disease; Drug Targeting; Genes; Glycoproteins; Goals; Healthcare; Hepatic; Hepatitis; Hepatitis C; Hepatitis C virus; Hepatocyte; Human; In Vitro; Infection; Intervention; Investigation; Knock-out; Knowledge; Life Cycle Stages; Light; Literature; Liver; Liver diseases; Modeling; Molecular; Mus; Pathway interactions; Play; Pluripotent Stem Cells; Population; Predisposition; Primary carcinoma of the liver cells; Process; Property; Proteins; RNA Interference; RNA Viruses; Reporting; Research; Resistance; Role; Specificity; Stem Cell Development; Stem cells; System; Therapeutic Intervention; Transplantation; Tropism; Vaccines; Validation; Viral; Viral hepatitis; Virus; Work; cofactor; complement C2a; design; drug development; in vivo; insight; liver function; liver transplantation; mouse model; new therapeutic target; novel; overexpression; pathogen; permissiveness; prevent; programs; prophylactic; public health relevance; research study; stem cell differentiation; stem cell technology; viral resistance

DESCRIPTION (provided by applicant): The broad, long-term goal of our program is to advance knowledge of virus-host cell interactions by characterizing cellular cofactors essential for hepatitis C virus (HCV) infection. HCV infects 170 million people worldwide and is major cause of hepatocellular carcinoma. Understanding how this virus interacts with the host cell (e.g., what makes a cell susceptible to HCV infection) is of critical importance to the development of diagnostics, antiviral drugs, and a prophylactic vaccine. Our previous studies of pluripotent stem cells and directed hepatic differentiation demonstrated a correlation between hepatic specification during the differentiation process and permissiveness to HCV infection. Characterization of the cellular transition from nonpermissive to permissive revealed candidate genes that are involved in regulating viral susceptibility. One of these cellular factors interacts with HCV glycoprotein E1 and plays an essential role in the HCV life cycle. The experiments proposed here will determine the molecular determinants of HCV permissiveness during hepatic development and identify new cellular cofactors and potential drug targets for HCV therapy. The following experiments will be performed: (1) Suppression of putative proviral factors and overexpression of putative antiviral factors in stem cells. These will be followed by hepatic differentiation and HCV infection of the differentiated human hepatocyte-like cells (DHHs). (2) Mechanistic studies of a novel proviral factor whose role has been validated by our preliminary studies. Specific steps of the HCV life cycle will be examined, and the interaction between viral and host protein will be dissected. (3) In vivo experiments designed to assess the potential of genetically modified DHHs to repopulate mouse liver and confer HCV resistance to the chimeric liver. The results of the proposed studies will not only provide significant insights into the properties of the host cells that govern HCV susceptibility but also reveal new therapeutic targets for antiviral intervention.

描述（由申请人提供）：我们程序的广泛长期目标是通过表征对乙型肝炎病毒（HCV）感染所必需的细胞辅助因子来提高病毒宿主相互作用的知识。 HCV在全球范围内感染了1.7亿人，是肝细胞癌的主要原因。了解该病毒如何与宿主细胞相互作用（例如，使细胞容易受到HCV感染的原因）对于诊断，抗病毒药物和预防性疫苗的发展至关重要。我们先前对多能干细胞和指示肝分化的研究表明，分化过程中的肝规范与HCV感染的允许性之间存在相关性。从非疗法到允许的候选基因的细胞过渡的表征，这些基因参与调节病毒易感性。这些细胞因子之一相互作用 使用HCV糖蛋白E1，并且在HCV生命周期中起着至关重要的作用。此处提出的实验将确定肝发育过程中HCV允许性的分子决定因素，并确定新的细胞辅助因子和HCV治疗的潜在药物靶标。将进行以下实验：（1）抑制假定病毒因子和干细胞中假定抗病毒因子的过表达。这些将是分化的人肝细胞样细胞（DHHS）的肝分化和HCV感染。 （2）一种新的病毒因子的机械研究，其作用已通过我们的初步研究证实。将检查HCV生命周期的特定步骤，并将解剖病毒和宿主蛋白之间的相互作用。 （3）旨在评估转基因DHHS重新填充小鼠肝脏并赋予HCV耐药性的体内实验。拟议的研究的结果不仅将为控制HCV易感性的宿主细胞的性质提供重大见解，而且还揭示了用于抗病毒干预的新的治疗靶标。