NIH: Spatial Models of Intrahepatic Hepatitis Virus Propagation in Humans

NIH：人类肝内肝炎病毒传播的空间模型

• 批准号: 10463188
• 负责人: ASHWIN BALAGOPAL
• 金额: $ 91.42万
• 依托单位: TRIAD NATIONAL SECURITY, LLC
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-02-07 至 2027-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10463188
• 关键词: Address; Antigens; Antiviral Agents; Antiviral Therapy; Biological Markers; Biology; Blood; Cell Nucleus; Cells; Chromosomes; Chronic; Chronic Hepatitis B; Circular DNA; Cirrhosis; Clinical; Clinical Markers; Collaborations; Complex; Data; Data Set; Development; Disease; Disease model; Distant; Equilibrium; Genetic Materials; Genetic Transcription; Genome; HIV; HIV/HCV; Hepatitis; Hepatitis B; Hepatitis B Surface Antigens; Hepatitis B Virus; Hepatitis C co-infection; Hepatitis C virus; Hepatitis Viruses; Hepatocyte; Human; Immunotherapy; Infection; Infection prevention; Interruption; Island; Knowledge; Link; Liver; Liver diseases; Measurement; Measures; Medicine; Modeling; Nucleic Acids; Organ; Organism; Paper; Pathology; Persons; Phase; Phylogenetic Analysis; Primary carcinoma of the liver cells; Process; Production; Proliferating; Proteins; Publishing; Recurrence; Research; Residual state; Sampling; Stream; Techniques; Testing; Therapeutic; Time; Tissues; Translational Research; Trees; United States National Institutes of Health; Vaccines; Variant; Viral; Viral Genome; Viral Pathogenesis; Virion; Virus; Virus Diseases; Virus Replication; Whole Organism; anti-PD-1; clinically relevant; co-infection; design; end stage liver disease; extracellular; genomic RNA; improved; innovation; insight; intrahepatic; liver biopsy; liver transplantation; mathematical model; multi-scale modeling; novel; repository; single cell analysis; single cell sequencing; success; tool; tumorigenesis; viral DNA; viral RNA; virus genetics; Address; Antigens; Antiviral Agents; Antiviral Therapy; Biological Markers; Biology; Blood; Cell Nucleus; Cells; Chromosomes; Chronic; Chronic Hepatitis B; Circular DNA; Cirrhosis; Clinical; Clinical Markers; Collaborations; Complex; Data; Data Set; Development; Disease; Disease model; Distant; Equilibrium; Genetic Materials; Genetic Transcription; Genome; HIV; HIV/HCV; Hepatitis; Hepatitis B; Hepatitis B Surface Antigens; Hepatitis B Virus; Hepatitis C co-infection; Hepatitis C virus; Hepatitis Viruses; Hepatocyte; Human; Immunotherapy; Infection; Infection prevention; Interruption; Island; Knowledge; Link; Liver; Liver diseases; Measurement; Measures; Medicine; Modeling; Nucleic Acids; Organ; Organism; Paper; Pathology; Persons; Phase; Phylogenetic Analysis; Primary carcinoma of the liver cells; Process; Production; Proliferating; Proteins; Publishing; Recurrence; Research; Residual state; Sampling; Stream; Techniques; Testing; Therapeutic; Time; Tissues; Translational Research; Trees; United States National Institutes of Health; Vaccines; Variant; Viral; Viral Genome; Viral Pathogenesis; Virion; Virus; Virus Diseases; Virus Replication; Whole Organism; anti-PD-1; clinically relevant; co-infection; design; end stage liver disease; extracellular; genomic RNA; improved; innovation; insight; intrahepatic; liver biopsy; liver transplantation; mathematical model; multi-scale modeling; novel; repository; single cell analysis; single cell sequencing; success; tool; tumorigenesis; viral DNA; viral RNA; virus genetics

Summary Significance: Hepatitis B virus (HBV) infects ~260 million people worldwide and is the leading cause of liver disease and hepatocellular carcinoma (HCC) in the world. Despite an effective vaccine for infection prevention, chronic hepatitis B virus infection (CHB) is difficult to cure once established. The viral genetic material enters the infected cell's nucleus generating a stable covalently closed-circular DNA (cccDNA) molecule or a linear form that integrates into the host cell's chromosome (iDNA). Existing treatments have little effect on cccDNA and as a result have poor success rates. Treatment interruption often leads to rebound of HBV DNA and recurrent hepatitis. In addition, even with treatment, liver disease in CHB often progresses to decompensated cirrhosis requiring liver transplantation. This process is exacerbated by several viral co-infections, such as with HIV or hepatitis C virus (HCV). Thus, improved therapeutics for CHB cure are required. Innovation: Our proposal is innovative in three aspects. i) access to precious and unique samples of liver tissue; ii) using state-of-the-art experimental techniques for single cell analyses and contemporaneous blood biomarkers; iii) development of new multi-scale models of HBV infection informed by our rich data sets, which are designed for clinical usefulness. Approach: HBV has a complex lifecycle comprising intracellular and extracellular phases. Our team has developed tools to interrogate the intracellular phase of HBV at the single-hepatocyte level. We have made preliminary observations of HBV in single cells but have only recently begun to link the intracellular and extracellular phases of HBV. Thus, we are poised to uncover key links in HBV biology between blood and liver. Using an integrated approach, we propose developing models of HBV that characterize replication in the organism uniting our understanding of intracellular and extracellular phases of the virus. Our specific aims are: i) To develop integrated models of intracellular and extracellular HBV replication, measuring and modeling a comprehensive set of HBV-related biomarkers in the blood and at the single-cell level in the liver, in untreated and treated CHB; and ii) To develop models of HBV spatial spread and persistence in the liver, using single-cell viral sequencing of HBV to make spatial linkages between genetically related HBV variants in the liver. These data and models will allow inferences about local versus non-local HBV spread. Specifically, we will test if clonal proliferation of HBV infected hepatocytes may contribute to continued persistence of infected cells, which has important implications not only for cure research but also for HCC oncogenesis. Impact: As multiple novel compounds for HBV treatment are in development, the timely insights generated in this proposal will fill gaps in our understanding of HBV replication and pathogenesis. Previous efforts by us and others in modeling HCV and HIV were important in the development of successful therapies for those infections. We expect the new data and models generated here to have the same impact in advancing HBV therapy.

概括 意义：丙型肝炎病毒（HBV）在全球感染了约2.6亿人，是肝脏的主要原因 世界上疾病和肝细胞癌（HCC）。尽管有有效的预防感染疫苗，但 慢性肝炎病毒感染（CHB）一旦建立就难以治愈。病毒遗传物质进入 感染的细胞核产生稳定的共价闭合DNA（CCCDNA）分子或线性 整合到宿主细胞染色体（IDNA）中的形式。现有治疗对CCCDNA几乎没有影响 因此，成功率很差。治疗中断通常会导致HBV DNA和 复发性肝炎。此外，即使接受治疗，CHB中的肝病也经常发展为代偿性 需要肝移植的肝硬化。几种病毒共同感染，例如，此过程加剧了 HIV或丙型肝炎病毒（HCV）。因此，需要改善CHB治疗的治疗剂。 创新：我们的建议在三个方面具有创新性。 i）获得肝脏的珍贵样本 组织; ii）使用最先进的实验技术进行单细胞分析和同期血液 生物标志物； iii）开发由我们丰富的数据集知之语的新型HBV感染模型 专为临床实用性而设计。 方法：HBV具有复杂的生命周期，包括细胞内和细胞外相。我们的团队有 开发了在单羊皮细胞水平上询问HBV的细胞内相的工具。我们做了 单个细胞中HBV的初步观察结果，但直到最近才开始联系细胞内和 HBV的细胞外相。因此，我们准备发现血液和肝脏之间HBV生物学的关键联系。 使用集成方法，我们提出了开发HBV模型，以表征在 有机体结合了我们对病毒细胞内和细胞外相的理解。我们的具体目的是： i）开发细胞内和细胞外HBV复制，测量和测量的集成模型 对血液中的HBV相关生物标志物进行建模，并在肝脏中的单细胞水平 未经治疗和处理的CHB； ii）开发肝脏中HBV空间扩散和持久性的模型， 使用HBV的单细胞病毒测序来在遗传相关的HBV变体之间建立空间连接 肝脏。这些数据和模型将允许推断局部与非本地HBV扩散。具体来说， 我们将测试HBV感染的肝细胞的克隆扩散是否可能导致继续持续 受感染的细胞，这不仅对治愈研究具有重要意义，而且对HCC肿瘤发生也具有重要意义。 影响：随着多种用于HBV治疗的新型化合物正在开发中，在 该建议将在我们对HBV复制和发病机理的理解中填补空白。我们以前的努力 对HCV和HIV进行建模的其他人在开发成功疗法方面很重要 感染。我们希望这里生成的新数据和模型在推进HBV方面产生相同的影响 治疗。