Modeling human hepatotropic infections in complex tissue organoids

描述（由申请方提供）：人类肝脏是几种重要人类病原体的储存库，包括乙型肝炎病毒（HBV）和丙型肝炎病毒（HCV）以及疟原虫属，所有这些都代表了严重的全球健康问题。仅HBV和HCV就慢性感染全球约5亿人，每年死亡人数超过150万。慢性HBV和HCV感染可导致严重的健康后果，包括肝炎、肝硬化、肝衰竭和肝细胞癌。合并感染HBV和HCV是常见的，并导致预后显着恶化。HBV的预防性疫苗存在，但针对该病毒的治疗性治疗不可用。此外，据报道，一线抗病毒药物的HBV耐药性增加。HCV的疫苗尚未开发，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，但快速出现的耐药性表明需要额外的靶点和抗病毒药物的组合来进行有效控制。忠实地模拟肝脏生物学和对人类嗜肝性病原体的易感性的体外和体内系统的缺乏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和联合收割机组织工程与分子病毒学和人源化小鼠技术相结合，以创建平台，将促进基础病毒-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，并提供预测系统来测试药物和疫苗的有效性和毒性。具体来说，我们的目标是在微模式化的原代人肝细胞培养物（MPCCs）中表征HBV和HCV生物学和模型相关肝病-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中的HBV和HCV感染。由于慢性炎症在肝脏疾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，并最终使用人类造血系统受体重建的小鼠用于HBV/HCV允许的肝脏类器官。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：B型和C型肝炎病毒是人类肝病（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单感染和合并感染。研究结果将有助于开发更有效的治疗方法，以根除这些致命的病毒性疾病。

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.