Liver fibrosis and CD206+ macrophage accumulation are suppressed by anti-GM-CSF therapy

• Published in: JHEP reports Vol. 2; no. 1; p. 100062
• Main Authors: Tan-Garcia, Alfonso, Lai, Fritz, Yeong, Joe Poh Sheng, Irac, Sergio E., Ng, Pei Y., Msallam, Rasha, Lim, Jeffrey Chun Tatt, 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
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 01.02.2020; Elsevier
• Subjects: anti-GM-CSF neutralizing antibody; CD206+ macrophages; fibrosis; Gastroenterology and Hepatology; GM-CSF; HCV; Intrahepatic macrophages; NASH; TCR; TNFα; ALT; t-SNE; moMΦs; LPS; SVR; HIER; GM-CSF; FFPE; HSC; TSA; DAA; MS; LSM; HCC; TMA; NASH; PBMCs; Intrahepatic macrophages; RA; CD206+ macrophages; anti-GM-CSF neutralizing antibody; BAMBI; ICS; fibrosis; HCV; DC; alanine aminotransferase; BMP and Activin Membrane-bound Inhibitor; CD206 + macrophages; hepatocellular carcinoma; hepatic stellate cells; T cell receptor; monocyte-derived macrophage-like cells; rheumatoid arthritis; tyramide signal amplification; tumour necrosis factor-α; granulocyte-macrophage colony-stimulating factor; lipopolysaccharide; dendritic cell; non-alcoholic steatohepatitis; tissue microarray; heat-induced epitope retrieval; formalin-fixed paraffin-embedded; sustained virological response; peripheral blood mononuclear cells; liver stiffness measurement; t-distributed stochastic neighbour embedding; multiple sclerosis; intracellular cytokine staining; direct-acting antiviral; TNFα, tumour necrosis factor-α; DC, dendritic cell; RA, rheumatoid arthritis; BAMBI, BMP and Activin Membrane-bound Inhibitor; ICS, intracellular cytokine staining; SVR, sustained virological response; HIER, heat-induced epitope retrieval; t-SNE, t-distributed stochastic neighbour embedding; moMΦs, monocyte-derived macrophage-like cells; GM-CSF, granulocyte-macrophage colony-stimulating factor; PBMCs, peripheral blood mononuclear cells; LSM, liver stiffness measurement; ALT, alanine aminotransferase; FFPE, formalin-fixed paraffin-embedded; TMA, tissue microarray; NASH, non-alcoholic steatohepatitis; MS, multiple sclerosis; TSA, tyramide signal amplification; DAA, direct-acting antiviral; HSC, hepatic stellate cells; LPS, lipopolysaccharide; HCC, hepatocellular carcinoma; TCR, T cell receptor
• ISSN: 2589-5559; 2589-5559
• DOI: 10.1016/j.jhepr.2019.11.006

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 in vitro and in vivo 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 in vitro that monocytes converted to TNFα-producing CD206+ macrophage-like cells in response to bacterial products (lipopolysaccharide) in a GM-CSF-dependent manner, confirming the in vivo 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. [Display omitted] •GM-CSF and TNFα producing CD206+ macrophages accumulate in human fibrotic liver•Serum GM-CSF is increased in HCV+ patients with higher liver fibrosis•GM-CSF drives monocyte to CD206+ macrophage conversion•Anti-GM-CSF therapy suppresses liver fibrosis and CD206+ macrophage accumulation