Expression of NKG2D ligands is downregulated by β-catenin signalling and associates with HCC aggressiveness

Background & Aims: The NKG2D system is a potent immuno-surveillance mechanism in cancer, wherein the activating NK cell receptor (NKG2D) on immune cells recognises its cognate ligands on tumour cells. Herein, we evaluated the expression of NKG2D ligands in hepatocellular carcinoma (HCC), in both humans and mice, taking the genomic features of HCC tumours into account. Methods: The expression of NKG2D ligands (MICA, MICB, ULBP1 and ULBP2) was analysed in large human HCC datasets by Fluidigm TaqMan and RNA-seq methods, and in 2 mouse models (mRNA and protein levels) reproducing the features of both major groups of human tumours. Results: We provide compelling evidence that expression of the MICA and MICB ligands in human HCC is associated with tumour aggressiveness and poor patient outcome. We also found that the expression of ULBP1 and ULBP2 was associated with poor patient outcome, and was downregulated in CTNNB1-mutated HCCs displaying low levels of inflammation and associated with a better prognosis. We also found an inverse correlation between ULBP1/2 expression levels and the expression of beta-catenin target genes in patients with HCC, suggesting a role for beta-catenin signalling in inhibiting expression. We showed in HCC mouse models that beta-catenin signalling downregulated the expression of Rae-1 NKG2D ligands, orthologs of ULBPs, through TCF4 binding. Conclusions: We demonstrate that the expression of NKG2D ligands is associated with aggressive liver tumorigenesis and that the downregulation of these ligands by beta-catenin signalling may account for the less aggressive phenotype of CTNNB1-mutated HCC tumours. Lay summary: The NKG2D system is a potent immuno-surveillance mechanism in cancer. However, its role in hepato-cellular carcinoma development has not been widely investigated. Herein, we should that the expression of NKG2D ligands by tumour cells is associated with a more aggressive tumour subtype. (C) 2021 European Association for the Study of the Liver. Published by Elsevier B.V. All rights reserved.