Lactate Activates TGF-β/SNAIL Signaling to Drive M2 Macrophage Polarization and CD8+ T Cell Exhaustion in Breast Cancer.

Abstract

This study demonstrates that lactate promotes M2-like macrophage polarization by activating the TGF-β/SNAIL signaling axis, thereby weakening CD8⁺ T cell-mediated antitumor immunity and promoting breast cancer progression. In vitro experiments using bone marrow-derived macrophages (BMDMs) and THP-1 cells treated with 25 mM lactate revealed a marked increase in M2 markers (CD206, Arg-1, IL-10) and a reduction in M1 markers (iNOS, TNF-α, IL-12), confirmed by Western blotting and flow cytometry. RNA-Seq analysis identified TGF-β/SNAIL pathway activation, with increased TGFBR1/2 expression, Smad2/3 phosphorylation, and PI3K/AKT pathway enrichment. Functional studies revealed that lactate-polarized M2 macrophages impaired CD8⁺ T cell cytotoxicity (reduced IFN-γ, GzmB, PRF1; elevated PD-1, Tim-3) and disrupted mitochondrial metabolism. In vivo validation using a breast cancer xenograft model showed that lactate treatment increased tumor growth and angiogenesis (VEGF/CD31⁺), while TGF-β inhibition (SB431542) reversed these effects. Mechanistically, lactate-induced TGF-β/SNAIL signaling promoted EMT in cancer cells and created an immunosuppressive TME. These findings establish lactate as a critical metabolic regulator that coordinates macrophage polarization and T cell exhaustion through the TGF-β/SNAIL axis, highlighting this pathway as a promising therapeutic target for breast cancer immunotherapy.