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Methylmalonic acid induces metabolic abnormalities and exhaustion in CD8+ T cells to suppress anti-tumor immunity

Oncogene volume 44, pages 105–114 (2025)Cite this article

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Abstract

Systemic levels of methylmalonic acid (MMA), a byproduct of propionate metabolism, increase with age and MMA promotes tumor progression via its direct effects in tumor cells. However, the role of MMA in modulating the tumor ecosystem remains to be investigated. The proliferation and function of CD8+ T cells, key anti-tumor immune cells, declines with age and in conditions of vitamin B12 deficiency, which are the two most well-established conditions that lead to increased systemic levels of MMA. Thus, we hypothesized that increased circulatory levels of MMA would lead to a suppression of CD8+ T cell immunity. Treatment of primary CD8+ T cells with MMA induced a dysfunctional phenotype characterized by robust immunosuppressive transcriptional reprogramming and marked increases in the expression of the exhaustion regulator, TOX. Accordingly, MMA treatment upregulated exhaustion markers in CD8+ T cells and decreased their effector functions, which drove the suppression of anti-tumor immunity in vitro and in vivo. Mechanistically, MMA-induced CD8+ T cell exhaustion was associated with a suppression of NADH-regenerating reactions in the TCA cycle and concomitant defects in mitochondrial function. Thus, MMA has immunomodulatory roles, thereby highlighting MMA as an important link between aging, immune dysfunction, and cancer.

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Fig. 1: MMA impairs activation and effector functions of primary CD8+ T cells.
Fig. 2: MMA-treated CD8+ T cells exhibit TCA cycle abnormalities and defective oxidative phosphorylation.
Fig. 3: MMA promotes a state of T cell exhaustion and the induction of TOX.
Fig. 4: MMA impairs CD8+ T cell cytotoxicity and augments tumor penetrance.

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Data availability

Data will be made available upon reasonable request to ana.dasilvagomes@moffitt.org.

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Acknowledgements

We would like to thank members of the DeNicola lab (Moffitt) for helpful feedback and support; Dr. Brian Ruffell and members of the Ruffell lab (Moffitt) for kindly sharing LLC-ZsGreen and LLC-OVA-ZsGreen cells as well as helpful advice; members of Dr. Javier Pinilla-Ibarz lab (Moffitt) for kindly sharing human PBMCs; and Drs. Alfred Zippelius (University of Basel) and Massimo Broggini (Instituto di Ricerche Farmacologiche Mario Negri IRCCS) for kindly sharing KP1.9 cells. We would also like to thank the Moffitt Cancer Center/USF Comparative Medicine Program for animal care and staff members of the Flow Cytometry, Proteomics and Metabolomics, and Analytic Microscopy Core facilities at the H. Lee Moffitt Cancer Center & Research Institute, an NCI designated Comprehensive Cancer Center (P30-CA076292). RSH is supported by the T32 CA233399 program. JLC is supported by the Cortner-Couch Endowed Chair for Cancer Research from the University of South Florida School of Medicine and by P01-CA250984. The mass spectrometry work was partially funded by NIH grants 5P01CA120964 (JMA) and 5P30CA006516 (JMA). JFG was supported by consecutive FWO postdoctoral fellowships. SMF acknowledges funding from Stichting tegen Kanker. This work was directly supported by the Pathway to Independence Award from NCI (R00CA218686; R00CA218686‐04S1). APG and the Gomes Laboratory are also supported by the New Innovator Award from OD/NIH (DP2AG0776980), an American Cancer Society Research Scholar Award (RSG-22-164-01-MM), the NIA (R21AG083720), the NCI (R01CA279023), the Florida Health Department Bankhead-Coley Research Program (24B03), METAvivor, the Florida Breast Cancer Research Foundation, and the Phi Beta Psi Sorority. Schematic figures were created using BioRender.com.

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Authors and Affiliations

  1. Department of Molecular Oncology, H. Lee Moffitt Cancer Center & Research Institute, Tampa, FL, 33612, USA

    Joanne D. Tejero, Stanislav Drapela, Didem Ilter, Devesh Raizada, Felicia Lazure, Hossein Kashfi & Ana P. Gomes

  2. Department of Molecular Medicine, University of South Florida Morsani College of Medicine, Tampa, FL, 33612, USA

    Joanne D. Tejero

  3. Department of Tumor Microenvironment and Metastasis, H. Lee Moffitt Cancer Center & Research Institute, Tampa, FL, 33612, USA

    Rebecca S. Hesterberg & John L. Cleveland

  4. Proteomics and Metabolomics Core, H. Lee Moffitt Cancer Center & Research Institute, Tampa, FL, 33612, USA

    Min Liu

  5. Department of Immunology, H. Lee Moffitt Cancer Center & Research Institute, Tampa, FL, 33612, USA

    Leonardo Silvane & Dorina Avram

  6. Laboratory of Cellular Metabolism and Metabolic Regulation, VIB-KU Leuven Center for Cancer Biology, VIB, Herestraat 49, 3000, Leuven, Belgium

    Juan Fernández-García & Sarah-Maria Fendt

  7. Laboratory of Cellular Metabolism and Metabolic Regulation, Department of Oncology, KU Leuven and Leuven Cancer Institute (LKI), Herestraat 49, 3000, Leuven, Belgium

    Juan Fernández-García & Sarah-Maria Fendt

  8. Department of Medicine, Beth Israel Deaconess Medical Center and Harvard Medical School, Boston, MA, 02215, USA

    John M. Asara

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Contributions

JDT contributed to experiment design and execution, data analysis, preparation of figures, and manuscript writing. RSH and SD contributed to experiment design and execution, data analysis, and interpretation. DI prepared the RNA for the RNA-seq experiment and performed its analysis and contributed to immunofluorescent tissue staining and lung cancer cell death experiments. SD, DR, FL, and HK contributed to tissue collection and processing of mouse experiments. JF-G and S-MF contributed to methodology and identification of MMA-regulated metabolism. ML performed the mass spectrometry analysis of MMA in mouse serum. LS and DA provided critical reagents. JLC contributed to project supervision, interpretation of results, and provided critical reagents. JA performed the metabolomic analysis. APG contributed to project conception and design, supervision, data analysis, data interpretation, funding acquisition, and manuscript writing. All authors have reviewed and approved the manuscript.

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Correspondence to Ana P. Gomes.

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Tejero, J.D., Hesterberg, R.S., Drapela, S. et al. Methylmalonic acid induces metabolic abnormalities and exhaustion in CD8+ T cells to suppress anti-tumor immunity. Oncogene 44, 105–114 (2025). https://doi.org/10.1038/s41388-024-03191-1

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