Developing a renewable and dissectible human liver for the study of HBV/HCV infection

开发可再生、可解剖的人类肝脏用于研究 HBV/HCV 感染

• 批准号: 10686216
• 负责人: Xianfang Wu
• 金额: $ 48.3万
• 依托单位: CLEVELAND CLINIC LERNER COM-CWRU
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-18 至 2027-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10686216
• 关键词: 3-Dimensional; Acceleration; Acute Liver Failure; Address; Affect; Age; Antiviral Agents; Area; CRISPR/Cas technology; Cell Communication; Cell Culture Techniques; Cell Differentiation process; Cell Line; Cells; Cellularity; Chronic Hepatitis B; Cirrhosis; Clinical; Coculture Techniques; Complex; DNA sequencing; Development; Diagnostic; Disease; Disease Progression; Endothelial Cells; Epidemiology; Fibrosis; Foundations; Future; Genes; Genetic; Genetic Variation; Goals; Hepatic; Hepatic Stellate Cell; Hepatitis A; Hepatitis B; Hepatitis B Infection; Hepatitis B Therapy; Hepatitis B Vaccines; Hepatitis B Virus; Hepatitis C; Hepatitis C Therapy; Hepatitis C co-infection; Hepatitis C virus; Hepatitis D; Hepatitis E virus; Hepatitis Viruses; Hepatocyte; Heterogeneity; Human; IL18 gene; In Vitro; Individual; Infection; Kupffer Cells; Link; Liver; Liver diseases; Macrophage; Mediating; Methods; Modeling; Molecular; Natural Immunity; Outcome; Patients; Persons; Population; Primary carcinoma of the liver cells; Reporting; Research; Risk; Sampling; Severities; Single Nucleotide Polymorphism; System; Technology; Time; Tissue Engineering; Translating; Treatment outcome; United States; Viral; Viremia; Virus; Work; aging population; career; cell type; chronic infection; chronic liver disease; co-infection; disease phenotype; epigenomics; genetic variant; genome wide association study; hemodynamics; high risk; human model; human pluripotent stem cell; improved; in vivo; individual variation; infection risk; innovation; insight; liver transplantation; mortality; novel; post-doctoral training; response; stem; stem cells; three dimensional cell culture; tool; transcriptomics; transmission process

Project Summary HBV and HCV infections are among the leading causes of chronic liver disease worldwide. In the United States, it is estimated that over 850 thousand people are currently infected with HBV and more than 2.4 million for HCV. Despite the availability of highly effective HBV vaccine and HCV treatment, the mortality and the burden associated with HBV and HCV are nevertheless increasing as individuals with existing infections advance into more advanced stages including fibrosis, cirrhosis, and HCC. Moreover, HCV-infected patients, even though cured of viremia, remain at a significantly elevated risk for advanced liver diseases. Due to their shared modes of transmission and epidemiological features, HBV and HCV frequently coexist in patients in endemic areas or among subjects at high risk of infection. Coinfection is usually more complex than monoinfection, leading to an accelerated disease progression and complicated viral interaction for their treatment. Despite the severity of HBV and HCV infections, the mechanisms by which they lead to liver disease, of how coinfection causes an increased severity and risk for complications, and of how HBV and HCV interact in the liver of coinfected patients that may lead to reactivation of HBV upon the cure of HCV remains largely unclear. This stems, in part, from the lack of relevant human models that recapitulate key disease phenotypes and permit detailed mechanistic studies that could answer these questions. The primary research goal of this proposal is to establish a novel, high fidelity multicellular in vitro liver model to address the aforementioned gaps. In this system, we plan to coculture human pluripotent stem cells (hPSCs)-derived hepatocytes, hepatic stellate cells, macrophages, and endothelial cells. Unlike existing traditional cell culture models, we propose to create an innovative three dimensional platform that mimics the liver’s multicellularity and microenvironment, by culturing these four cell types in a configuration that resembles the liver’s in vivo spatial organization, its hemodynamics, and its cell-cell interaction. To distinguish cell type- specific responses, we will develop methods to purify individual cell types for downstream analysis. With this multicellular culture platform, we will pursue our long-range objectives, across which we capitalize on the unique features of this novel platform to address questions that cannot be adequately answered with any existing in vitro human-relevant system. These include the mechanisms of continued disease progression following the cure of HCV infection, an accelerated disease progression with HBV/HCV coinfection, and the interaction between HBV/HCV that leads to HBV reactivation following the cure of HCV infection (Aim 1). In Aim 2, we will combine hPSCs with gene editing to understand how genetic variants affect the course of disease and treatment following HBV and HCV infection. These studies will not only provide new insights that could inform development of effective diagnostics and treatments for HBV/HCV infection, but also lead to technology advance that open up new avenues for future studies focusing on other liver diseases.

项目概要 HBV 和 HCV 感染是全球慢性肝病的主要原因之一。在美国， 据估计，目前有超过85万人感染乙肝病毒，超过240万人感染丙肝病毒。 尽管有高效的乙型肝炎疫苗和丙型肝炎治疗，但死亡率和负担 然而，随着现有感染者进入新的阶段，与 HBV 和 HCV 相关的疾病仍在增加。 更晚期阶段包括纤维化、肝硬化和肝癌。此外，HCV 感染者即使 病毒血症治愈后，患晚期肝病的风险仍然显着升高。由于他们共享模式 从传播和流行病学特征来看，乙型肝炎和丙型肝炎经常在流行地区或地区的患者中同时存在。 属于感染高危人群。混合感染通常比单一感染更复杂，导致 加速疾病进展并使其治疗变得复杂的病毒相互作用。 尽管 HBV 和 HCV 感染很严重，但它们导致肝病的机制仍 合并感染如何导致并发症的严重程度和风险增加，以及 HBV 和 HCV 在感染中如何相互作用 合并感染患者的肝脏在 HCV 治愈后可能导致 HBV 重新激活的机制尚不清楚。 这在一定程度上是由于缺乏相关的人类模型来概括关键疾病表型并允许 可以回答这些问题的详细机制研究。 该提案的主要研究目标是建立一种新型的、高保真度的体外多细胞肝脏 模型来解决上述差距。在这个系统中，我们计划共培养人类多能干细胞 (hPSC) 衍生的肝细胞、肝星状细胞、巨噬细胞和内皮细胞。与现有的不同 传统的细胞培养模型，我们建议创建一个创新的三维平台来模仿 通过以类似的配置培养这四种细胞类型，研究肝脏的多细胞性和微环境 肝脏的体内空间组织、血流动力学和细胞间相互作用。区分细胞类型 - 针对具体的反应，我们将开发纯化单个细胞类型以进行下游分析的方法。 借助这个多细胞培养平台，我们将追求我们的长期目标，并利用这些目标 利用这个新颖平台的独特功能来解决任何无法充分回答的问题 现有的体外人体相关系统。这些包括疾病持续进展的机制 HCV 感染治愈后，HBV/HCV 合并感染加速疾病进展，以及 HBV/HCV 之间的相互作用导致 HCV 感染治愈后 HBV 再激活（目标 1）。瞄准 2、我们将hPSC与基因编辑结合起来，了解基因变异如何影响病程以及 HBV 和 HCV 感染后的治疗。这些研究不仅会提供新的见解，可以为 开发针对 HBV/HCV 感染的有效诊断和治疗方法，同时也带来技术进步 这为未来关注其他肝脏疾病的研究开辟了新的途径。