Targeting MMP9 in CTNNB1 mutant hepatocellular carcinoma restores CD8⁺ T cell-mediated antitumour immunity and improves anti-PD-1 efficacy

Objective The gain of function (GOF) CTNNB1 mutations (CTNNB1(GOF)) in hepatocellular carcinoma (HCC) cause significant immune escape and resistance to anti-PD-1. Here, we aimed to investigate the mechanism of CTNNB1(GOF) HCC-mediated immune escape and raise a new therapeutic strategy to enhance anti-PD-1 efficacy in HCC.Design RNA sequencing was performed to identify the key downstream genes of CTNNB1(GOF) associated with immune escape. An in vitro coculture system, murine subcutaneous or orthotopic models, spontaneously tumourigenic models in conditional gene-knock-out mice and flow cytometry were used to explore the biological function of matrix metallopeptidase 9 (MMP9) in tumour progression and immune escape. Single-cell RNA sequencing and proteomics were used to gain insight into the underlying mechanisms of MMP9.Results MMP9 was significantly upregulated in CTNNB1(GOF) HCC. MMP9 suppressed infiltration and cytotoxicity of CD8(+) T cells, which was critical for CTNNB1(GOF) to drive the suppressive tumour immune microenvironment (TIME) and anti-PD-1 resistance. Mechanistically, CTNNB1(GOF) downregulated sirtuin 2 (SIRT2), resulting in promotion of beta-catenin/lysine demethylase 4D (KDM4D) complex formation that fostered the transcriptional activation of MMP9. The secretion of MMP9 from HCC mediated slingshot protein phosphatase 1 (SSH1) shedding from CD8(+) T cells, leading to the inhibition of C-X-C motif chemokine receptor 3 (CXCR3)-mediated intracellular of G protein-coupled receptors signalling. Additionally, MMP9 blockade remodelled the TIME and potentiated the sensitivity of anti-PD-1 therapy in HCC.ConclusionsCTNNB1(GOF) induces a suppressive TIME by activating secretion of MMP9. Targeting MMP9 reshapes TIME and potentiates anti-PD-1 efficacy in CTNNB1(GOF) HCC.