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

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：乙型肝炎病毒 (HBV) 感染全球约 2.6 亿人，是导致肝病的主要原因 世界范围内的疾病和肝细胞癌（HCC）。尽管有有效的疫苗来预防感染， 慢性乙型肝炎病毒感染（CHB）一旦形成就很难治愈。病毒遗传物质进入 受感染细胞的细胞核产生稳定的共价闭环 DNA (cccDNA) 分子或线性 整合到宿主细胞染色体（iDNA）中的形式。现有的治疗方法对 cccDNA 影响不大 因此成功率很低。治疗中断通常会导致 HBV DNA 反弹， 复发性肝炎。此外，即使经过治疗，慢性乙型肝炎患者的肝病也常常会进展为失代偿性。 肝硬化需要肝移植。几种病毒的共同感染会加剧这一过程，例如 HIV 或丙型肝炎病毒 (HCV)。因此，需要改进治疗慢性乙型肝炎的疗法。 创新：我们的方案在三个方面具有创新性。 i) 获得珍贵且独特的肝脏样本 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) 根据我们丰富的数据集开发新的多尺度乙肝病毒感染模型， 专为临床用途而设计。 方法：HBV 具有复杂的生命周期，包括细胞内和细胞外阶段。我们的团队有 开发了在单肝细胞水平上检测 HBV 细胞内阶段的工具。我们已经做了 对单细胞中的 HBV 进行了初步观察，但直到最近才开始将细胞内和 HBV 的细胞外期。因此，我们准备揭示血液和肝脏之间 HBV 生物学的关键联系。 我们建议使用综合方法开发 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阶段的理解。我们的具体目标是： i) 开发细胞内和细胞外 HBV 复制、测量和检测的综合模型 对血液中和肝脏单细胞水平上的一组全面的 HBV 相关生物标志物进行建模， 未经治疗和已治疗的慢性乙型肝炎； ii) 开发乙肝病毒空间传播和肝脏持久性模型， 使用 HBV 的单细胞病毒测序来建立遗传相关的 HBV 变异之间的空间联系 肝脏。这些数据和模型将有助于推断本地与非本地乙肝病毒的传播。具体来说， 我们将测试 HBV 感染的肝细胞的克隆增殖是否可能有助于持续存在 被感染的细胞，这不仅对治疗研究而且对 HCC 肿瘤发生也具有重要意义。 影响：随着多种治疗乙肝病毒的新型化合物正在开发中，及时产生的见解 该提议将填补我们对乙型肝炎病毒复制和发病机制理解的空白。我们之前的努力和 HCV 和 HIV 建模中的其他人对于开发成功的治疗方法非常重要 感染。我们预计这里生成的新数据和模型对推进乙型肝炎具有相同的影响 治疗。