Co-targeting TGF-β and PD-L1 sensitizes triple-negative breast cancer to experimental immunogenic cisplatin-eribulin chemotherapy doublet
Laura Kalfeist, … , Emeric Limagne, Sylvain Ladoire
Laura Kalfeist, … , Emeric Limagne, Sylvain Ladoire
Published July 1, 2025
Citation Information: J Clin Invest. 2025;135(13):e184422. https://doi.org/10.1172/JCI184422.
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Research Article Immunology Oncology

Co-targeting TGF-β and PD-L1 sensitizes triple-negative breast cancer to experimental immunogenic cisplatin-eribulin chemotherapy doublet

Abstract

In preclinical mouse models of triple-negative breast cancer (TNBC), we show that a combination of chemotherapy with cisplatin (CDDP) and eribulin (Eri) was additive from an immunological point of view and was accompanied by the induction of an intratumoral immune and inflammatory response favored by the immunogenic cell death induced by CDDP, as well as by the vascular and tumor stromal remodeling induced by each chemotherapy. Unexpectedly, despite the favorable immune context created by this immunomodulatory chemotherapy combination, our models remained refractory to the addition of anti–PD-L1 immunotherapy. These surprising observations led us to discover that CDDP chemotherapy was simultaneously responsible for the production of TGF-β by several populations of cells present in tumors, which favored the emergence of different subpopulations of immune cells and cancer-associated fibroblasts characterized by immunosuppressive properties. Accordingly, co-treatment with anti–TGF-β restored the immunological synergy between this immunogenic doublet of chemotherapy and anti–PD-L1 in a CD8-dependent manner. Translational studies revealed the unfavorable prognostic effect of the TGF-β pathway on the immune response in human TNBC, as well as the ability of CDDP to induce this cytokine also in human TNBC cell lines, thus highlighting the clinical relevance of targeting TGF-β in the context of human TNBC treated with chemoimmunotherapy.

Authors

Laura Kalfeist, Fanny Ledys, Stacy Petit, Cyriane Poirrier, Samia Kada Mohammed, Loïck Galland, Valentin Derangère, Alis Ilie, David Rageot, Romain Aucagne, Pierre-Simon Bellaye, Caroline Truntzer, Marion Thibaudin, Mickaël Rialland, François Ghiringhelli, Emeric Limagne, Sylvain Ladoire

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Figure 4

The CDDP-Eri doublet induces TGF-β–mediated immunoresistance.

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The CDDP-Eri doublet induces TGF-β–mediated immunoresistance. (A) 4T1 tu...
(A) 4T1 tumor–bearing mice were treated with CDDP, Eri, or both. Tumors were collected at days 4, 8, and 14. At day 8, total tumor mRNA was analyzed for immunosuppressive gene expression by NanoString. The heatmap on the left shows normalized expression, and the heatmap on the right shows statistical significance (n = 4/group). P values were determined by 2-way ANOVA. (B) Tgfb1 expression was measured by qPCR at days (D) 4, 8, and 14 (n ≥6/group). (C and D) 4T1 cells were treated with Eri (50 nM), CDDP (4 μM), CDDP-Eri, or left untreated. Tgfb1 (C) and Serpin1 (D) expression was analyzed by qPCR at 24 hours and 48 hours (2 experiments, n = 3/experiment). (E) 4T1 cells were pretreated with galunisertib (10, 100, and 1,000 nM) for 2 hours, and then treated with CDDP (4 μM), TGF-β (2 ng/mL), or left untreated for 48 hours. Serpin1 expression was analyzed by qPCR (2 experiments, n = 2/experiment). (F) Under the same conditions as in C, Smad2/3 phosphorylation [pSmad2 (Ser465/467), pSmad3 (Ser423/425)] was analyzed by Western blotting. Heatmap represents the phosphorylated/total ratio (results are from 1 of 2 experiments). (G and H) 4T1 tumor–bearing mice were treated as in A. At day 4, immunosuppressive cells were sorted, and Tgfb1 expression was measured by qPCR (G). MyCAF-related genes were analyzed in CAFs (H) (n ≥3/group). (I) At day 8, αSMA+ CAF proportions were measured by flow cytometry. (J and K) αSMA+ cells were quantified by IHC (J), and collagen deposition was analyzed by Masson’s trichrome (MTC) staining (K) (n ≥7/group). Images are shown again in Supplemental Figure 4I. (L–N) Mo-MDSC (L), TAM2 (M), and Treg (N) proportions were assessed by flow cytometry (n = 5/group). Box plots represent the mean ± SEM. *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001, by 1-way ANOVA (C–E) and 2-way ANOVA (B, G, H, and L–N).