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.
View: Text | PDF
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

×

Figure 3

Anti–TIM-3 Ab treatment reduces Tc exhaustion in vitro.

Options: View larger image (or click on image) Download as PowerPoint
Anti–TIM-3 Ab treatment reduces Tc exhaustion in vitro. (A–G) Tc isolate...
(A–G) Tc isolated from spleens (C57BL/6 mice) were continuously exposed to TCR stimulation (αCD3/CD28 beads) in vitro for 14 days in RPMI medium supplemented with 10% FBS, 1% penicillin/streptomycin, and 30 U/mL of mIL-2 to obtain highly activated/exhausted Tc. Cells were treated every second day with 10 μg/mL of isotype or anti–TIM-3 Ab. (A) Representative plots showing the proportion of TIM-3+PD-1+ cells among viable CD4+ and CD8+ Tc treated with isotype or anti–TIM-3, determined by FC. Relative TIM-3 protein expression in CD4+ Tc (B) and in CD8+ Tc (C) based on MFI for n = 6 replicates for each condition. P values were calculated using Wilcoxon’s signed-rank test. (D and E) Proportion (%) of TIM-3+PD-1+ within all CD4+ Tc (D) and within all CD8+ Tc (E). Data are represented as mean ± SEM for isotype Ab (n = 6) and anti–TIM-3 Ab (n = 6). P values were calculated using Mann-Whitney U test. (F) Relative TOX protein expression as relative MFI determined by FC in CD4+ Tc and (G) representative FC staining of TOX expression in CD4+ Tc for both isotype Ab (blue line) and anti–TIM-3 Ab treatment (red line). (H) Relative TOX protein expression as relative MFI determined by FC in CD8+ Tc and (I) representative staining of TOX expression in CD8+ Tc for both isotype Ab (blue line) and anti–TIM-3 Ab treatment (red line). Data are represented as mean ± SEM of n = 5 isotype treatment or n = 5 anti–TIM-3 treatment. P values were calculated using Wilcoxon’s signed-rank test.