Abstract
Although most patients with relapsed/refractory B-cell acute lymphoblastic leukemia (B-ALL) receiving CD19-targeted chimeric antigen receptor (CAR) T cell therapy achieve remission, loss of CAR T cell functionality and subsequent relapse remains an unmet therapeutic need. Herein, we apply an integrative approach to study the immunometabolism of pre- and post-infusion CD19-CAR T cells of patients with relapsed/refractory B-ALL. Pre-infusion CAR T cells of long-term responders (LTR) have increased oxidative phosphorylation, fatty acid oxidation, and pentose phosphate pathway activities, higher mitochondrial mass, tighter cristae, and lower mTOR expression compared to products of short-term responders. Post-infusion CAR T cells in bone marrow (BM) of LTR have high immunometabolic plasticity and mTOR-pS6 expression supported by the BM microenvironment. Transient inhibition of mTOR during manufacture induces metabolic reprogramming and enhances anti-tumor activity of CAR T cells. Our findings provide insight into immunometabolic determinants of long-term response and suggest a therapeutic strategy to improve long-term remission.
Data availability
The RNA sequencing data generated in this study are available at the Gene Expression Omnibus (GEO) repository of the National Center for Biotechnology Information under accession code GSE298663. Metabolomic profiles are available at the NIH Common Fund’s National Metabolomics Data Repository (NMDR) website, the Metabolomics Workbench42 (https://www.metabolomicsworkbench.org, Study IDs: ST003963, ST003964, ST003966. Supplementary information, including Supplementary Figs. 1–8 and Supplementary Data files 1–6 are provided with the online version of this paper. All other datasets generated during and/or analyzed during this study are available from the corresponding author on request. Source data are provided with this paper.
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Acknowledgements
Research reported in this publication included work performed in the Integrative Genomics Core and the Integrated Mass Spectrometry Shared Resource supported by the National Cancer Institute of the National Institutes of Health under grant number P30CA033572. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. L.G. acknowledges support from NIH K12 grant no. 5K12CA001727–29, the Hyundai Hope on Wheels Young Investigator Award, the Margaret E. Early Medical Research Trust Award, the Schwartz Accelerator Fund, the Norman and Sadie Lee Foundation, and the Albert and Bettie Sacchi Foundation. The authors thank Chunyan Zhang for her meticulous technical support throughout the preparation of this manuscript.
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L.G., S.J.F., and X. Wang. conceived and conceptualized the study. L.G. performed the in vitro and in vivo experiments, analyzed the data, and wrote the manuscript. E.R.H. performed cytometry data analysis and generated plots. J.W., B.G., R.U., V.V., and R.E. performed in vitro and in vivo experiments and analyzed data. K.V.P., N.P.H., and P.P. performed and analyzed the mass spectrometry experiments. J.S. and J.L.F. performed the extracellular flux experiments. R.Z.N. and Z.L. performed transmission electron microscope experiments. D.W., E.T., and R.K.G. analyzed transmission electron microscope and mass spectrometry data. M.H.C. and X. Wu. analyzed RNA data. T.M., S.B., J.R.W., J.P., M.C.C., D.N., and I.A. collected and analyzed patient data. L.G. and M.C.C. wrote the original draft. All authors reviewed and edited the final manuscript. L.G., X. Wang., and S.J.F. provided resources, acquired funding, and supervised the study.
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Goldberg, L., Haas, E.R., Wu, J. et al. Immunometabolic determinants of long-term response in leukemia patients receiving CD19 CAR T cell therapy. Nat Commun (2026). https://doi.org/10.1038/s41467-026-69857-4
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DOI: https://doi.org/10.1038/s41467-026-69857-4
