Modeling human hepatotropic infections in complex tissue organoids

在复杂组织类器官中模拟人类嗜肝感染

• 批准号: 7935261
• 负责人: SANGEETA N. BHATIA
• 金额: $ 112.63万
• 依托单位: ROCKEFELLER UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-30 至 2014-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7935261
• 关键词: Antiviral Agents; Biology; Cessation of life; Chronic; Cirrhosis; Clinical; Complex; Development; Disease Progression; HCV Vaccine; Health; Hematopoietic System; Hepatitis; Hepatitis B; Hepatitis B Vaccines; Hepatitis B Virus; Hepatitis C; Hepatitis C virus; Hepatocyte; Human; Immune system; In Vitro; Infection; Inflammation; Laboratories; Light; Liver; Liver Failure; Liver diseases; Malignant Neoplasms; Methodology; Modeling; Molecular Virology; Mono-S; Mus; Organoids; Pathogenesis; Peptide Hydrolases; Pharmaceutical Preparations; Plasmodium; Play; Polymerase; Predisposition; Primary carcinoma of the liver cells; Reporting; Resistance; Role; Structure; System; Technology; Therapeutic Intervention; Tissue Engineering; Tissues; Toxic effect; Transplantation; Virus; Virus Diseases; drug testing; effective therapy; global health; in vivo; inhibitor/antagonist; interdisciplinary approach; novel; outcome forecast; pathogen; public health relevance; reconstitution; tissue culture; vaccine development; vaccine efficacy; virology; virus virus interaction

DESCRIPTION (provided by applicant): The human liver serves as the reservoir for several important human pathogens, including hepatitis B (HBV) and C viruses (HCV) and Plasmodium species, all of which represent serious global health concerns. HBV and HCV alone chronically infect an estimated 500 million people worldwide, with annual deaths totaling more than 1.5 million. Chronic HBV and HCV infections can have severe health consequences, including hepatitis, cirrhosis, liver failure, and hepatocellular carcinoma. Co-infection with HBV and HCV is common, and leads to a significantly worse prognosis. A preventative vaccine for HBV exists, but curative treatments targeting the virus are not available. Furthermore, increasing HBV resistance has been reported to first-line antiviral drugs. A vaccine for HCV has not yet been developed, and, while HCV-specific protease and polymerase inhibitors are showing promise in early clinical development, rapid emergence of resistance indicates that additional targets and combinations of antivirals will be needed for effective control. The scarcity of in vitro and in vivo systems that faithfully mimic liver biology and susceptibility to human hepatotropic pathogens has severely hampered drug and vaccine development. Here, we propose to take an interdisciplinary approach and combine tissue engineering with molecular virology and humanized mouse technology to create platforms that will facilitate studies of basic virus-host and virus-virus interactions, promote understanding of the mechanisms of liver disease progression, and provide predictive systems to test drug and vaccine efficacy and toxicity. Specifically, we aim to characterize HBV and HCV biology and model associated liver disease in micropatterned primary human hepatocyte cultures (MPCCs) - a breakthrough technology that was recently developed in Dr. Sangeeta Bhatia's laboratory. We will extend this methodology to develop three-dimensional liver organoids, and investigate HBV and HCV infection in mice transplanted with these structures. Since chronic inflammation plays a significant role in liver disease progression, we aim to incorporate components of the immune system in static and dynamic tissue culture systems and ultimately to use mice reconstituted with a human hematopoietic system recipients for HBV/HCV-permissive liver organoids. Through the development and use of these platforms, we hope to shed light on HBV and HCV virology and pathogenesis, and to uncover novel avenues for therapeutic intervention. PUBLIC HEALTH RELEVANCE: Hepatitis B and C viruses are leading causes of human liver disease including cancer. This proposal aims to create and utilize novel primary human hepatocyte tissue culture systems and complex 3D liver organoids to study HCV and HBV mono- and co-infections. The resulting findings should aid development of more effective treatments aimed at eradicating these deadly viral diseases.

描述（由申请人提供）：人类肝脏是多种重要人类病原体的储存库，包括乙型肝炎（HBV）和丙型肝炎病毒（HCV）以及疟原虫物种，所有这些都代表着严重的全球健康问题。据估计，仅 HBV 和 HCV 就长期感染全世界 5 亿人，每年死亡人数总计超过 150 万人。慢性乙型肝炎和丙型肝炎病毒感染可造成严重的健康后果，包括肝炎、肝硬化、肝功能衰竭和肝细胞癌。乙型肝炎病毒和丙型肝炎病毒合并感染很常见，并导致预后明显较差。乙型肝炎病毒已有预防性疫苗，但尚无针对该病毒的治疗方法。此外，据报道，一线抗病毒药物的乙肝病毒耐药性不断增加。 HCV 疫苗尚未开发出来，虽然 HCV 特异性蛋白酶和聚合酶抑制剂在早期临床开发中显示出希望，但耐药性的迅速出现表明需要额外的靶标和抗病毒药物组合才能有效控制。忠实模拟肝脏生物学的体外和体内系统的稀缺以及对人类嗜肝病原体的敏感性严重阻碍了药物和疫苗的开发。在这里，我们建议采取跨学科的方法，将组织工程与分子病毒学和人源化小鼠技术相结合，创建平台，以促进基本病毒-宿主和病毒-病毒相互作用的研究，促进对肝病进展机制的理解，并提供测试药物和疫苗功效和毒性的预测系统。具体来说，我们的目标是在微图案原代人肝细胞培养物 (MPCC) 中表征 HBV 和 HCV 生物学特征并建立相关肝脏疾病模型，这是 Sangeeta Bhatia 博士实验室最近开发的一项突破性技术。我们将扩展这种方法来开发三维肝脏类器官，并研究移植这些结构的小鼠的 HBV 和 HCV 感染情况。由于慢性炎症在肝病进展中起着重要作用，我们的目标是将免疫系统的组成部分纳入静态和动态组织培养系统中，并最终使用与人类造血系统受体重建的小鼠来获得 HBV/HCV 许可的肝脏类器官。通过这些平台的开发和使用，我们希望阐明 HBV 和 HCV 病毒学和发病机制，并发现治疗干预的新途径。 公共卫生相关性：乙型和丙型肝炎病毒是人类肝脏疾病（包括癌症）的主要原因。该提案旨在创建和利用新型原代人肝细胞组织培养系统和复杂的 3D 肝脏类器官来研究 HCV 和 HBV 单一感染和混合感染。由此产生的发现应该有助于开发更有效的治疗方法，旨在根除这些致命的病毒性疾病。