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IMMUNOTHERAPY

Improved CAR-T cell activity associated with increased mitochondrial function primed by galactose

Leukemia (2024)Cite this article

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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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Fig. 1: Mitochondrial parameters of clinical CAR-T cell products of six pediatric ALL patients.
Fig. 2: CD19 CAR-T cell metabolism and activity in glucose and galactose.
Fig. 3: Functional properties of CAR-T cells are impacted by galactose.
Fig. 4: Production in galactose improves CAR-T cell function.

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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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Authors and Affiliations

  1. Cell Therapy Lab, Sheba Medical Center, Tel Hashomer, Israel

    Golda Gross, Suha Alkadieri, Amilia Meir, Yarden Aharoni-Tevet, Shahar Ben Yosef & Elad Jacoby

  2. Faculty of Medicinal & Health Sciences, Tel Aviv University, Tel Aviv, Israel

    Golda Gross, Suha Alkadieri, Yarden Aharoni-Tevet, Angi Zenab, Amos Toren & Elad Jacoby

  3. Ella Institute of Immuno-Oncology, Sheba Medical Center, Tel Hashomer, Israel

    Orit Itzhaki

  4. Bert Strassburger Metabolic Center for Preventive Medicine, Sheba Medical Center, Tel Hashomer, Israel

    Angi Zenab, Liat Shbiro & Tal Yardeni

  5. Division of Pediatric Hematology and Oncology, The Edmond and Lily Safra Children’s Hospital, Sheba Medical Center, Tel Hashomer, Israel

    Amos Toren & Elad Jacoby

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  1. Golda Gross
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Contributions

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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Correspondence to Elad Jacoby.

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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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