Primary cell culture models of HIV/HBV co-infection

项目摘要 全世界约有2.5亿人慢性感染B型肝炎病毒（HBV） 他们有患肝硬化和肝细胞癌的风险。乙肝相关性疾病导致的生命损失 每年并发症约82万人。与此同时，人类免疫缺陷病毒（HIV）感染 全世界约有3800万人。其中，全球7.4%和高度流行地区的15-28%也是 感染HBV。艾滋病毒合并感染造成严重的健康负担，因为它们加速了肝脏的进展 肝硬化和肝癌，其机制仍不清楚。 HBV感染的慢性化与共价闭合环状DNA的稳定性和维持有关 HBV基因组的核形式是cccDNA（cccDNA）。当HBV感染肝脏中的人肝细胞时， cccDNA在这些细胞中建立，并且旨在消除或消除它的策略被认为是有效的。 功能性治疗的解决方案。目前针对慢性HBV的治疗包括核苷类似物和干扰素 α。虽然核苷类似物不直接影响cccDNA，但接受干扰素治疗的有限数量的患者 α或核苷类似物可以解决感染并实现治愈。对全球HBV治愈的需求是迫切的， 这也是为什么一些方法和研究化合物旨在治愈。 HBV是一种仅感染人肝细胞的专性嗜肝病毒。HBV研究受到阻碍 通过有限的细胞培养系统，包括HBV受体NTCP过表达后的肝癌细胞， 原代人肝细胞。后者是生理上最相关的细胞，但它们的可用性和 原代细胞的固有局限性使得难以使用它们来重现完整的HBV生命周期。到 为此，我们开发了基于原代人肝细胞的细胞培养系统， 人源化小鼠，其有效地支持长期HBV感染和传播。此外，我们开发了一个系统， 从HBV感染的人源化小鼠中分离这些细胞，这为研究提供了独特的机会。 具体地，细胞被HBV慢性感染，因此携带高水平的cccDNA，这允许HBV感染。 体外模拟慢性HBV。在这里，我们建议使用这些创新的肝细胞系统， 与CD 4 + T细胞共培养，用于在生理相关原代细胞培养中研究HIV/HBV共感染 系统.总的来说，通过我们提出的工作，我们将深入了解HIV合并感染对HBV生命周期的影响 和肝细胞反应，我们将建立一个平台，以确定新的抗病毒策略， 有效的乙肝病毒治疗

Project Summary There are approximately a quarter of a billion people worldwide chronically infected with hepatitis B virus (HBV) who are at risk of developing liver cirrhosis and hepatocellular carcinoma. The loss of life due to HBV-related complications each year is about 820,000 people. At the same time human immunodeficiency virus (HIV) infects about 38 million people worldwide. Among those, 7.4% globally and 15-28% in highly endemic areas are also infected with HBV. HIV co-infections pose a serious health burden as they accelerate the progression to liver cirrhosis and liver cancer with mechanisms that are still not well understood. Chronicity of HBV infections is linked to the stability and maintenance of the covalently closed circular DNA (cccDNA), which is the nuclear form of the HBV genome. As HBV infects human hepatocytes in the liver, the cccDNA is established in these cells and strategies that aim to inactivate or eliminate it are considered the solution to a functional cure. Current treatments against chronic HBV include nucleoside analogs and interferon α. While nucleoside analogs do not directly affect the cccDNA, a limited number of patients that receive interferon α or nucleoside analogs can resolve the infection and achieve a cure. The need for a global HBV cure is urgent, which is also why several approaches and investigational compounds aim for a cure. HBV is an exclusive hepatotropic virus that only infects human hepatocytes. HBV studies have been hindered by limited cell culture systems that include hepatoma cells after overexpression of the HBV receptor NTCP and primary human hepatocytes. The latter are the most physiologically relevant cells, but their availability and the inherent limitations of primary cells makes it difficult to use them to recapitulate the complete HBV lifecycle. To this end, we have developed cell culture systems based on primary human hepatocytes, isolated from humanized mice, that efficiently support long-term HBV infection and spread. Moreover, we developed a system of isolating these cells from HBV-infected humanized mice which provides unique opportunities for research. Specifically, cells are chronically infected with HBV and therefore carry high levels of cccDNA, which allows for modeling chronic HBV in vitro. Here we propose to use these innovative hepatocyte systems to establish robust co-cultures with CD4+ T cells for studying HIV/HBV co-infections in physiologically relevant primary cell culture systems. Overall, with our proposed work we will gain insights on the effects of HIV co-infection on HBV lifecycle and hepatocyte responses and we will establish a platform to identify novel antiviral strategies with the goal for a functional HBV cure.