BTK signaling drives CD1dhiCD5+ regulatory B-cell differentiation to promote pancreatic carcinogenesis

Abstract

The immune microenvironment of pancreatic ductal adenocarcinoma (PDA) is comprised of a heterogeneous population of cells that are critical for disease evolution. Prominent among these are the specialized CD1d^hiCD5⁺ regulatory B (B_reg) cells that exert a pro-tumorigenic role by promoting tumor cell proliferation. Dissecting the molecular pathways regulating this immune sub-population can thus be valuable for uncovering potential therapeutic targets. Here, we investigate Bruton's tyrosine kinase (BTK), a key B-cell kinase, as a potential regulator of CD1d^hiCD5⁺ B_reg differentiation in the pancreatic tumor microenvironment. Treatment of cytokine-induced B cells in vitro with the high specificity BTK inhibitor Tirabrutinib inhibited CD1d^hiCD5⁺ B_reg differentiation and production of IL-10 and IL-35, essential mediators of B_reg immunosuppressive functions. The BTK signaling pathway was also found to be active in vivo in PanIN-associated regulatory B cells. Tirabrutinib treatment of mice bearing orthotopic Kras^G12D-pancreatic lesions severely compromised stromal accumulation of the CD1d^hiCD5⁺ B_reg population. This was accompanied by an increase in stromal CD8⁺IFNγ⁺ cytotoxic T cells and significant attenuation of tumor cell proliferation and PanIN growth. Our results uncover a novel role for BTK in regulating CD1d^hiCD5⁺ B_reg differentiation and emphasize its potential as a therapeutic target for pancreatic cancer.