Abstract
CD19 CAR-T cells have led to durable remissions in patients with refractory B-cell malignancies; nevertheless, most patients eventually relapse in the long term. Many interventions aimed at improving current products have been reported, with a subset of them focusing on a direct or indirect link to the metabolic state of the CAR-T cells. We assessed clinical products from an ongoing clinical trial utilizing CD19-28z CAR-T cells from patients with acute lymphoblastic leukemia. CAR-T clinical products leading to a complete response had significantly higher mitochondrial function (by oxygen consumption rate) irrespective of mitochondrial content. Next, we replaced the carbon source of the media from glucose to galactose to impact cellular metabolism. Galactose-containing media increased mitochondrial activity in CAR-T cells, and improved in in-vitro efficacy, without any consistent phenotypic change in memory profile. Finally, CAR-T cells produced in galactose-based glucose-free media resulted in increased mitochondrial activity. Using an in-vivo model of Nalm6 injected mice, galactose-primed CAR-T cells significantly improved leukemia-free survival compared to standard glucose-cultured CAR-T cells. Our results prove the significance of mitochondrial metabolism on CAR-T cell efficacy and suggest a translational pathway to improve clinical products.
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Data availability
All data is available upon request. A pre-print version was posted and can be accessed here: BIORXIV/2023/559091.
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Acknowledgements
The authors thank the production team at the Ella Institute of Immuno-oncology who produced the clinical CAR-T cells. This work was carried out in partial requirements for GG and SA’s MSc degree (Faculty of Medicinal and Health Sciences, Tel Aviv University).
Funding
This work was funded by the Dotan center for hematologic malignancies grant and Israel Cancer Association grant 20240064 (EJ) and NIH grant 5R01CA259635 (TY).
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GG, TY and EJ conceived the initial study plan. GG, SA, AM, YAT, SBY AZ, LS, TY and EJ performed experiments and analyzed the data. AT, TY and EJ supervised the work. GG, TY and EJ wrote the manuscript draft. All authors approved the manuscript.
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Gross, G., Alkadieri, S., Meir, A. et al. Improved CAR-T cell activity associated with increased mitochondrial function primed by galactose. Leukemia (2024). https://doi.org/10.1038/s41375-024-02257-z
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DOI: https://doi.org/10.1038/s41375-024-02257-z