Tan-Garcia Alfonso, Lai Fritz, Sheng Yeong Joe Poh, Irac Sergio E, Ng Pei Y, Msallam Rasha, Tatt Lim Jeffrey Chun, Wai Lu-En, Tham Christine Y L, Choo Su P, Lim Tony, Young Dan Y, D'Ambrosio Roberta, Degasperi Elisabetta, Perbellini Riccardo, Newell Evan, Le Bert Nina, Ginhoux Florent, Bertoletti Antonio, Chen Qingfeng, Dutertre Charles-Antoine
Program in Emerging Infectious Diseases, Duke-NUS Medical School, 8 College Road, Singapore 169857, Singapore.
Humanised Mouse Unit, Institute of Molecular and Cell Biology, ASTAR, 61 Biopolis Drive, Singapore 138673, Singapore.
JHEP Rep. 2019 Dec 6;2(1):100062. doi: 10.1016/j.jhepr.2019.11.006. eCollection 2020 Feb.
BACKGROUND & AIMS: Chronic liver inflammation leads to fibrosis and cirrhosis and is associated with an accumulation of intrahepatic TNFα-secreting CD206 macrophages, which may participate in maintaining chronic liver disease in a GM-CSF-dependent manner. We aimed to elucidate the exact role of GM-CSF in the development and progression of chronic liver disease.
Liver immunohistochemistry and serum quantification were performed in patients with viral and non-viral-related liver disease to compare CD206 monocyte/macrophages, fibrosis and GM-CSF. This was followed by functional validations and in humanised mice.
Using multiplex immunofluorescence and histo-cytometry, we show that highly fibrotic livers had a greater density of CD206 macrophages that produced more TNFα and GM-CSF in the non-tumour liver regions of patients with hepatocellular carcinoma (n = 47), independent of aetiology. In addition, the absolute number of CD206 macrophages strongly correlated with the absolute number of GM-CSF-producing macrophages. In non-HCC chronic HCV patients (n = 40), circulating GM-CSF levels were also increased in proportion to the degree of liver fibrosis and serum viral titres. We then demonstrated that monocytes converted to TNFα-producing CD206 macrophage-like cells in response to bacterial products (lipopolysaccharide) in a GM-CSF-dependent manner, confirming the normalisation of serum GM-CSF concentration following oral antibiotic treatment observed in HBV-infected humanised mice. Finally, anti-GM-CSF neutralising antibody treatment reduced intrahepatic CD206 macrophage accumulation and abolished liver fibrosis in HBV-infected humanised mice.
While the direct involvement of CD206 macrophages in liver fibrosis remains to be demonstrated, these findings show that GM-CSF may play a central role in liver fibrosis and could guide the development of anti-GM-CSF antibody-based therapy for the management of patients with chronic liver disease.
Liver fibrosis is a major driver of liver disease progression. Herein, we have shown that granulocyte-macrophage colony-stimulating factor (GM-CSF) plays an important role in the development of liver fibrosis. Our findings support the use of anti-GM-CSF neutralising antibodies for the management of patients with chronic liver disease resulting from both viral and non-viral causes.
慢性肝脏炎症会导致肝纤维化和肝硬化，且与肝内分泌肿瘤坏死因子α（TNFα）的CD206巨噬细胞的积累有关，这些巨噬细胞可能以粒细胞-巨噬细胞集落刺激因子（GM-CSF）依赖的方式参与维持慢性肝病。我们旨在阐明GM-CSF在慢性肝病发生发展过程中的确切作用。
对病毒性和非病毒性肝病患者进行肝脏免疫组化及血清定量分析，以比较CD206单核细胞/巨噬细胞、肝纤维化和GM-CSF的情况。随后在人源化小鼠中进行功能验证。
通过多重免疫荧光和组织细胞计数法，我们发现，在肝细胞癌患者（n = 47）的非肿瘤肝区域，高度纤维化的肝脏中CD206巨噬细胞密度更高，这些巨噬细胞产生更多的TNFα和GM-CSF，且与病因无关。此外，CD206巨噬细胞的绝对数量与产生GM-CSF的巨噬细胞的绝对数量密切相关。在非肝癌慢性丙型肝炎病毒（HCV）患者（n = 40）中，循环GM-CSF水平也与肝纤维化程度和血清病毒滴度成比例增加。然后我们证明，单核细胞在GM-CSF依赖的方式下对细菌产物（脂多糖）作出反应，转化为分泌TNFα的CD206巨噬细胞样细胞，这证实了在感染乙肝病毒的人源化小鼠中口服抗生素治疗后血清GM-CSF浓度恢复正常。最后，抗GM-CSF中和抗体治疗减少了感染乙肝病毒的人源化小鼠肝内CD206巨噬细胞的积累，并消除了肝纤维化。
虽然CD206巨噬细胞在肝纤维化中的直接作用仍有待证实，但这些发现表明GM-CSF可能在肝纤维化中起核心作用，并可为基于抗GM-CSF抗体的疗法的开发提供指导，用于治疗慢性肝病患者。
肝纤维化是肝病进展的主要驱动因素。在此，我们表明粒细胞-巨噬细胞集落刺激因子（GM-CSF）在肝纤维化的发生发展中起重要作用。我们的研究结果支持使用抗GM-CSF中和抗体来治疗由病毒和非病毒原因引起的慢性肝病患者。
