BACKGROUND AND AIMS HCV infection can be successfully managed with antiviral therapies; however, progression to chronic liver disease states, including NAFLD, is common. There is currently no reliable in vitro model for investigating host-viral interactions underlying the link between HCV and NAFLD; although liver organoids (LOs) show promise, they currently lack non-parenchymal cells, which are key to modeling disease progression. APPROACH AND RESULTS Here, we present a novel, multicellular LO model using a co-culture system of macrophages and LOs differentiated from the same human pluripotent stem cells (PSCs). The co-cultured macrophages shifted towards a Kupffer-like cell type, the liver-resident macrophages present in vivo, providing a suitable model for investigating NAFLD pathogenesis. With this multicellular Kupffer-like cell-containing LO (Kp-LO) model, we found that HCV infection led to lipid accumulation in LOs by upregulating host lipogenesis, which was more marked with macrophage co-culture. Reciprocally, long-term treatment of LOs with fatty acids (FAs) upregulated HCV amplification and promoted inflammation and fibrosis. Notably, in our Kp-LO model, the effects of three drugs for NASH that have reached phase 3 clinical trials exhibited consistent results with the clinical outcomes. CONCLUSIONS Taken together, we introduced a multicellular LO model consisting of hepatocytes, Kupffer-like cells, and HSCs, which recapitulated host-virus intercommunication and inter-cellular interactions. With this novel model, we present a physiologically relevant system for the investigation of NAFLD progression in HCV patients.

中文翻译：

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用于研究丙型肝炎病毒感染和非酒精性脂肪肝疾病进展的多细胞肝脏类器官模型。

背景和目的 HCV 感染可以通过抗病毒治疗成功控制；然而，进展为慢性肝病（包括 NAFLD）是很常见的。目前还没有可靠的体外模型来研究 HCV 和 NAFLD 之间的宿主-病毒相互作用；尽管肝脏类器官（LO）显示出前景，但它们目前缺乏非实质细胞，而非实质细胞是模拟疾病进展的关键。方法和结果在这里，我们提出了一种新颖的多细胞 LO 模型，该模型使用由相同的人类多能干细胞 (PSC) 分化而来的巨噬细胞和 LO 的共培养系统。共培养的巨噬细胞转向库普弗样细胞类型，即体内存在的肝脏巨噬细胞，为研究 NAFLD 发病机制提供了合适的模型。通过这种含有多细胞库普弗细胞的 LO (Kp-LO) 模型，我们发现 HCV 感染通过上调宿主脂肪生成导致 LO 中的脂质积累，这在巨噬细胞共培养中更为明显。相反，长期用脂肪酸 (FA) 治疗 LO 会上调 HCV 扩增并促进炎症和纤维化。值得注意的是，在我们的 Kp-LO 模型中，三种已达到 3 期临床试验的 NASH 药物的效果与临床结果表现出一致的结果。结论 综上所述，我们引入了由肝细胞、Kupffer 样细胞和 HSC 组成的多细胞 LO 模型，该模型概括了宿主与病毒之间的相互通讯和细胞间相互作用。通过这种新模型，我们提出了一个生理相关系统，用于研究 HCV 患者 NAFLD 进展。