Oncogene-induced TIM-3 ligand expression dictates susceptibility to anti–TIM-3 therapy in mice
Nana Talvard-Balland, et al.
Nana Talvard-Balland, et al.
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Research Article

Oncogene-induced TIM-3 ligand expression dictates susceptibility to anti–TIM-3 therapy in mice

Abstract

Leukemia relapse is a major cause of death after allogeneic hematopoietic cell transplantation (allo-HCT). We tested the potential of targeting T cell (Tc) immunoglobulin and mucin-containing molecule 3 (TIM-3) for improving graft-versus-leukemia (GVL) effects. We observed differential expression of TIM-3 ligands when hematopoietic stem cells overexpressed certain oncogenic-driver mutations. Anti–TIM-3 Ab treatment improved survival of mice bearing leukemia with oncogene-induced TIM-3 ligand expression. Conversely, leukemia cells with low ligand expression were anti–TIM-3 treatment resistant. In vitro, TIM-3 blockade or genetic deletion in CD8+ Tc enhanced Tc activation, proliferation, and IFN-γ production while enhancing GVL effects, preventing Tc exhaustion, and improving Tc cytotoxicity and glycolysis in vivo. Conversely, TIM-3 deletion in myeloid cells did not affect allogeneic Tc proliferation and activation in vitro, suggesting that anti–TIM-3 treatment–mediated GVL effects are Tc induced. In contrast to anti–programmed cell death protein 1 (anti–PD-1) and anti–cytotoxic T lymphocyte–associated protein 4 (anti–CTLA-4) treatment, anti–TIM-3-treatment did not enhance acute graft-versus-host disease (aGVHD). TIM-3 and its ligands were frequently expressed in acute myeloid leukemia (AML) cells of patients with post–allo-HCT relapse. We decipher the connections between oncogenic mutations found in AML and TIM-3 ligand expression and identify anti–TIM-3 treatment as a strategy for enhancing GVL effects via metabolic and transcriptional Tc reprogramming without exacerbation of aGVHD. Our findings support clinical testing of anti–TIM-3 Ab in patients with AML relapse after allo-HCT.

Authors

Nana Talvard-Balland, Lukas M. Braun, Karen O. Dixon, Melissa Zwick, Helena Engel, Alina Hartmann, Sandra Duquesne, Livius Penter, Geoffroy Andrieux, Lukas Rindlisbacher, Andrea Acerbis, Jule Ehmann, Christoph Köllerer, Michela Ansuinelli, Andres Rettig, Kevin Moschallski, Petya Apostolova, Tilman Brummer, Anna L. Illert, Markus A. Schramm, Yurong Cheng, Anna Köttgen, Justus Duyster, Hans D. Menssen, Jerome Ritz, Bruce R. Blazar, Melanie Boerries, Annette Schmitt-Gräff, Nurefsan Sariipek, Peter Van Galen, Joerg M. Buescher, Nina Cabezas-Wallscheid, Heike L. Pahl, Erika L. Pearce, Robert J. Soiffer, Catherine J. Wu, Luca Vago, Burkhard Becher, Natalie Köhler, Tobias Wertheimer, Vijay K. Kuchroo, Robert Zeiser

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Figure 7

Anti-human TIM-3 Ab sabatolimab enhances the GVL effect.

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Anti-human TIM-3 Ab sabatolimab enhances the GVL effect. (A) Kaplan-Meie...
(A) Kaplan-Meier plots showing mouse survival in the indicated groups. Rag2–/–Il2rg–/– recipient mice were injected i.v. with human-derived MOLM-13luc+ AML cells and human CD3+ Tc (from HLA-nonmatched healthy donors) and treated with vehicle (n = 10) or human anti–TIM-3 (n = 10). Results show 3 independent experiments, and P value was calculated using 2-sided Mantel-Cox test. (B) Representative images of bioluminescence imaging (BLI) of MOLM-13luc+ AML–bearing Rag2–/–Il2rg–/– recipient mice 21 days after injection of AML cells, following human Tc injection and vehicle or human anti–TIM-3 Ab treatment. (C) BLI signal quantification shows the expansion of AML cells over time. Data are represented as mean ± SEM from 3 independent experiments using 3 different healthy Tc donors. P values were calculated using 2-sided Mann-Whitney U test. (D and E) Kaplan-Meier plots showing mouse survival in the indicated groups. Rag2–/–Il2rg–/– recipient mice were injected i.v. with CD3-depleted primary AML cells (from PB at primary diagnosis) and treated with vehicle (n = 3) or human anti–TIM-3 (n = 3). Each survival curve represents 1 individual AML donor patient. P values were calculated using 2-sided Mantel-Cox test. (F) Representative Western blots showing the inhibition of phosphorylated FLT3 (pFLT3) (Tyr589/591) upon treatment with 3 different FLT3 inhibitors and loading control (β-actin) in MOLM-13 cells (FLT3-ITD). (G) Gal-9, (H) CEACAM1, and (I) TIM-3 protein expression as relative MFI was determined by FC upon the treatment with the indicated FLT3 inhibitors in FLT3-ITD–positive human cell line (MOLM-13) or FLT3 WT human cell line (Kasumi-1). Fold change is calculated in comparison with the treatment using DMSO (control treatment). Data are represented as mean ± SEM from 8 independent experiments using MOLM-13 cells and n = 5 independent experiments using Kasumi-1 cells. P values were calculated using ordinary 1-way ANOVA.