Oncogene-induced TIM-3 ligand expression dictates susceptibility to anti–TIM-3 therapy in mice
Nana Talvard-Balland, … , Vijay K. Kuchroo, Robert Zeiser
Nana Talvard-Balland, … , Vijay K. Kuchroo, Robert Zeiser
Published June 25, 2024
Citation Information: J Clin Invest. 2024;134(16):e177460. https://doi.org/10.1172/JCI177460.
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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 1

TIM-3 inhibition enhances GVL effects in different AML models.

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TIM-3 inhibition enhances GVL effects in different AML models. Murine pr...
Murine primary HSCs (C57BL/6) were transduced with the indicated oncogenes or gene fusions. RNA expression of (A) Lgals9 and (B) Ceacam1 was determined by quantitative PCR (qPCR). Gene expression was normalized to Hprt. Fold change was calculated in comparison with empty vector (dotted line). Results are represented as mean ± SEM from 3 independent experiments. P values were calculated using Kruskal-Wallis 1-way ANOVA multiple-comparisons test. (C–E) Kaplan-Meier plots showing mouse survival in the indicated groups. BALB/c recipient mice were injected i.v. with 7 × 104 oncogene-transduced HSCs and 5 × 106 allogeneic BM and/or Tc when indicated. All mice were treated i.p. with isotype Ab or anti–TIM-3 Ab as indicated. P values were calculated using 2-sided Mantel-Cox test. (F–I) RNA expression of (F) Havcr2, (G) Lgals9, (H) Ceacam1, and (I) Hmgb1 in WEHI-3B (n = 13) and in FLT3-ITD MLL-PTD cells (n = 6) was analyzed by qPCR. Gene expression is shown as percentage of Hprt. P values were calculated using an unpaired Student’s t test. (J and K) Kaplan-Meier plot showing mouse survival in the indicated groups. Recipient mice were injected i.v. with FLT3-ITD MLL-PTD (J) or WEHI-3B cells (K) and 5 × 106 allogeneic BM and/or Tc and treated with either isotype Ab or anti–TIM-3 Ab, as indicated. P value was calculated using 2-sided Mantel-Cox test. (K) Kaplan-Meier plots showing mouse survival in the indicated groups. BALB/c recipient mice were injected with WEHI-3B cells and 5 × 106 allogeneic BM and Tc and treated with isotype Ab (n = 10) or anti–TIM-3 Ab (n = 10). Data are represented as 2 independent experiments. P value was calculated using 2-sided Mantel-Cox test. (L) Data are represented as mean ± SEM of CD45+H-2Kb+ cell frequency in the BM at day 23 after allo-HCT. C57BL/6 recipient mice were injected with FLT3-ITD MLL-PTD cells and allogeneic BM and/or Tc and treated with isotype Ab (n = 10) or anti–TIM-3 Ab (n = 10). P values were calculated using 1-way ANOVA followed by Tukey’s multiple-comparisons test.