Pharmacologic LDH inhibition redirects intratumoral glucose uptake and improves antitumor immunity in solid tumor models
Svena Verma, Sadna Budhu, Inna Serganova, Lauren Dong, Levi M. Mangarin, Jonathan F. Khan, Mamadou A. Bah, Anais Assouvie, Yacine Marouf, Isabell Schulze, Roberta Zappasodi, Jedd D. Wolchok, Taha Merghoub
Svena Verma, Sadna Budhu, Inna Serganova, Lauren Dong, Levi M. Mangarin, Jonathan F. Khan, Mamadou A. Bah, Anais Assouvie, Yacine Marouf, Isabell Schulze, Roberta Zappasodi, Jedd D. Wolchok, Taha Merghoub
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Research Article Immunology Metabolism

Pharmacologic LDH inhibition redirects intratumoral glucose uptake and improves antitumor immunity in solid tumor models

Abstract

Tumor reliance on glycolysis is a hallmark of cancer. Immunotherapy is more effective in controlling glycolysis-low tumors lacking lactate dehydrogenase (LDH) due to reduced tumor lactate efflux and enhanced glucose availability within the tumor microenvironment (TME). LDH inhibitors (LDHi) reduce glucose uptake and tumor growth in preclinical models, but their impact on tumor-infiltrating T cells is not fully elucidated. Tumor cells have higher basal LDH expression and glycolysis levels compared with infiltrating T cells, creating a therapeutic opportunity for tumor-specific targeting of glycolysis. We demonstrate that LDHi treatment (a) decreases tumor cell glucose uptake, expression of the glucose transporter GLUT1, and tumor cell proliferation while (b) increasing glucose uptake, GLUT1 expression, and proliferation of tumor-infiltrating T cells. Accordingly, increasing glucose availability in the microenvironment via LDH inhibition leads to improved tumor-killing T cell function and impaired Treg immunosuppressive activity in vitro. Moreover, combining LDH inhibition with immune checkpoint blockade therapy effectively controls murine melanoma and colon cancer progression by promoting effector T cell infiltration and activation while destabilizing Tregs. Our results establish LDH inhibition as an effective strategy for rebalancing glucose availability for T cells within the TME, which can enhance T cell function and antitumor immunity.

Authors

Svena Verma, Sadna Budhu, Inna Serganova, Lauren Dong, Levi M. Mangarin, Jonathan F. Khan, Mamadou A. Bah, Anais Assouvie, Yacine Marouf, Isabell Schulze, Roberta Zappasodi, Jedd D. Wolchok, Taha Merghoub

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Figure 5

LDH inhibition improves antitumor T cell functions.

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LDH inhibition improves antitumor T cell functions. (A) Schematic depict...
(A) Schematic depicting tumor-killing assay with LDHi in which B16-YFP cells were treated with 20 μM LDHi or vehicle 24 hours apart and T cells were added 24 hours after the first LDHi treatment. (B) Quantified media glucose from killing assay coculture. (C) Flow cytometry quantification of 2-NBDG (MFI) in B16-YFP and CD8+ Pmel-1 T cells from killing assay cocultures 48 hours after last treatment. (D–F) (D) Quantified YFP+ tumor cells and (E) representative in vitro killing assay images of YFP+ tumor cells after 48 hours of coincubation with Pmel-1 CD8+ T cells as in A. (F) Corresponding quantified YFP+ tumor cells and percentages of tumor killing in the same conditions as above alongside vehicle supplemented with 10 mM glucose. (G) Quantification of killing of OVA257-264–pulsed live B16-YFP tumor cells by OVA-primed CD8+ T cells from OT1 transgenic mice upon 48 hours of coculture in the presence of LDHi (as indicated in A). E:T = 2:1, cocultured over 48 hours. (H) Schematic depicting in vitro Treg suppression assay with MACS column–sorted Tregs (CD4+CD25+ Regulatory T Cell Isolation Kit, mouse) cocultured with αCD3/αCD28-activated CTV-labeled syngeneic CD8+ T cells for 48 hours with the addition of conditioned media from B16 cells treated with 20 μM LDHi or vehicle or fresh media containing 10 mM glucose. (I) Percentage of suppression was calculated as percentage reduction in CD8+ T cell proliferation with respect to CD8+ T cells cultured alone in the same treatment and glucose conditions. Data show 1 representative experiment of 3 independent experiments (n = 3–4 technical replicates). All statistics produced by 2-way ANOVA with Bonferroni’s multiple-comparisons test implemented in GraphPad Prism. *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001. Data are represented as mean ± SEM.