Tumor-derived lactate inhibit the efficacy of lenvatinib through regulating PD-L1 expression on neutrophil in hepatocellular carcinoma

Abstract

BackgroundNeutrophils play a controversial role in tumor development. The function of programmed cell death-1 ligand (PD-L1+) neutrophils, however, may inhibit the cytotoxicity of anti-tumor immunity. In this study, we elucidate the stimulators of PD-L1+ neutrophils in tumor microenvironment (TME) and explore the optimal combination to enhance the effect of lenvatinib by inhibiting PD-L1+ neutrophils in hepatocellular carcinoma.MethodsNeutrophil infiltration after lenvatinib treatment was examined with RNA sequencing and multicolor flow cytometry analysis in patient samples, subcutaneous and orthotopic mouse models. Neutrophils and T cells were isolated from peripheral blood and tumor tissues and purified with magnetic beads for cytotoxicity assay. Metabolites and cytokines were detected by a biochemical analyzer manufactured by Yellow Springs Instrument (YSI) and proteome profiler cytokines array. In vitro screening of pathway inhibitors was used to identify possible candidates that could reduce PD-L1+ neutrophil infiltration. Further in vivo assays were used for verification.ResultsLenvatinib increased neutrophil recruitment by inducing CXCL2 and CXCL5 secretion in TME. After entering TME, neutrophils polarized toward N2 phenotype. PD-L1 expression was simultaneously upregulated. Thus, lenvatinib efficacy on tumor cells hindered. The increasing PD-L1+ neutrophils positively corelated with a suppressive T cell phenotype. Further investigation indicated that JAK/STAT1 pathway activated by immune-cell-derived interferon γ and MCT1/NF-kB/COX-2 pathway activated by high concentrations of tumor-derived lactate could induce PD-L1+ neutrophils. The latter could be significantly inhibited by COX-2 inhibitor celecoxib. Further in vivo assays verified that Celecoxib decreased the survival of lactate-stimulated PD-L1+ neutrophil and promoted the antitumor effect of lenvatinib.ConclusionsPD-L1+ neutrophils decrease T cell cytotoxicity. Tumor-derived lactate induces PD-L1 expression on neutrophils via MCT1/NF-κB/COX-2 pathway. Thus, COX-2 inhibitor could reduce PD-L1+ neutrophil and restore T cell cytotoxicity. This may provide a potent addition to lenvatinib.

Lenvatinib plus transarterial chemoembolization with or without immune checkpoint inhibitors for unresectable hepatocellular carcinoma: A review

“…Adachi et al held that the increase of PD-L1 will enhance the effect of PD-1 inhibitors (66). Lenvatinib also increased neutrophil and upregulated PD-L1 expression on neutrophils in the TME (77). In conclusion, the effect of lenvatinib on PD-L1 expression remains controversial and has not been finalized.…”

Dual FGFR and VEGFR inhibition synergistically restrain hexokinase 2-dependent lymphangiogenesis and immune escape in intrahepatic cholangiocarcinoma

“…6C). NF-Κb/Cox-2 pathway are reported to be associated with PD-L1 expression regulated by lactic acid [11]. The western blot results showed that lactate dehydrogenase A (LDHA), NF-Κb and Cox-2 expressions in LECs were signi cantly decreased after combined therapy (Fig.…”

“…Under steady-state conditions, LECs express high levels of T-cell inhibitory molecules including PD-L1 (CD274), which inducing apoptosis in tumorspeci c CD8 + T cells [9]. Many studies have revealed that lactic acid, the ultimate down-stream metabolite of anaerobic glycolysis, was responsible for driving PD-L1 expression [10,11]. Strikingly, lymphangiogenesis involving robust sprouting, migration and proliferation of LECs are heavily rely on anaerobic glycolysis for ATP production [12].…”

A Novel Lactate Metabolism-Related Gene Signature for Predicting Clinical Outcome and Tumor Microenvironment in Hepatocellular Carcinoma

“…High levels of lactate in the TME promote differentiation of tumor-associated macrophages to the M2 subtype, while activated macrophages facilitate tumor invasion through the CCL17/CCR4/mTORC1 signaling axis ( Zhang et al, 2021 ). Lactate-induced PD-L1 up-regulation on neutrophils impairs T cell cytotoxicity in HCC ( Deng et al, 2021 ). In addition, tumor cell-derived lactate induces the expression of GPR81 in dendritic cells via paracrine mode to inhibit the antigen presentation function of immune cells ( Brown et al, 2020 ).…”