Fibroblast growth factor inhibition by molecular-targeted agents mitigates immunosuppressive tissue microenvironment in hepatocellular carcinoma

Abstract

Background & aims Combination immunotherapy refers to the use of immune checkpoint inhibitors (ICI) and molecular-targeted agents (MTA), which have recently been approved for the treatment of advanced hepatocellular carcinoma (HCC). Owing to its relatively low antitumor effect (up to 30%), sequential therapy following ICIs treatment is required in patients with HCC. This study aimed to determine the impact of MTAs on the tumor immune microenvironment (TIME). Methods We established immune syngeneic orthotopic HCC mouse models using Hep-55.1C and Hep-53.4, and treated them with MTAs (lenvatinib, sorafenib, regorafenib, cabozantinib, and DC101 as anti-vascular endothelial growth factor receptor-2 antibodies, and AZD4547 as a fibroblast growth factor receptor (FGFR)-1/2/3/4 inhibitor) for 2 weeks. Subsequently, alterations in the TIME caused by MTAs were evaluated using immunohistochemistry (antibodies for CD3, CD8, Foxp3, Granzyme B, Arginase-1, NK1.1, F4/80, CD11c, PD-1, and PD-L1). We conducted RNA-seq analysis using lenvatinib- and AZD4547-treated tumors. To confirm the clinical relevance of these findings, we analyzed the transcriptome data of human HCC cells (MHCC-97H) treated with various concentrations of lenvatinib for 24 h using RNA-seq data from the Gene Expression Omnibus database. Results The number of Foxp3- and F4/80-positive cells in the TIME was decreased in many MTAs. Cabozantinib increased the numbers in NK1.1-, Granzyme B, and CD11c-positive cells. Lenvatinib and AZD4547 increased the number of CD8, Granzyme B, and PD-L1-positive cells. Gene ontology enrichment analysis revealed that lipid metabolism-related genes were downregulated by lenvatinib and AZD4547. In total, 161 genes downregulated by FGFR inhibition in rodent models overlapped with those downregulated by lenvatinib in human HCC cells. Conclusions In this study, we showed that cabozantinib activated the innate immune system, and lenvatinib and AZD4547, which commonly inhibit FGFR signaling, altered TIME to a hot immune state by downregulating lipid metabolism-related genes. These findings support the therapeutic use of combination immunotherapies.

Expression and Clinicopathological Relevance of Fibroblast Growth Factor Receptors in Canine Mammary Gland Tumors

“…Recently, FGFR5 lacking the intracellular tyrosine kinase domain, but serves to regulate excessive activation of the pathway by the ligand has been identified (Yue et al 2021). Activation of the receptors by ligand; Fibroblast growth factor (FGF) binding leads to the activation of a series of cascades, which are regulatory to cell growth, differentiation, migration and survival in cells derived from mesoderm and neuroectoderm (Haugsten et al 2010;Qin et al 2019;Suzuki et al 2023). Aberrant expression of FGFRs has been reported to be involved in carcinogenesis and progression of selected human neoplasia including breast cancer (Blanckaert et al 1998;Haugsten et al 2010;Kuroso et al 2010;Helsten et al 2016;Qin et al 2019;Wu et al 2022;Bou Zerdan et al 2023), with FGFR2 suggested as a breast cancer susceptibility gene (Meyer et al 2008;Sun et al 2012).…”