CAR signaling instructs divergent metabolic reprogramming and functional fates in αβ and γδ T cells
5 affiliations
- 1Department of Immunology, H. Lee Moffitt Cancer Center & Research Institute, Tampa, FL USA
- 2Proteomics & Metabolomics Core, H. Lee Moffitt Cancer Center & Research Institute, Tampa, FL USA
- 3Molecular Oncology, H. Lee Moffitt Cancer Center & Research Institute, Tampa, FL USA
- 4Department of Biostatistics and Bioinformatics, H. Lee Moffitt Cancer Center & Research Institute, Tampa, FL USA
- 5Department of Cutaneous Oncology, H. Lee Moffitt Cancer Center & Research Institute, Tampa, FL USA
Abstract
γδ T-cell-based immunotherapies have become relevant as alternatives to conventional αβ T-cell products, with preclinical data demonstrating tumor burden reduction and the mitigation of tumor-induced tissue damage. Given that most CAR constructs have been optimized for αβ T cells, we hypothesized that distinct T-cell types may require tailored CAR architectures to achieve optimal function. To test this hypothesis, we conducted a systematic comparative analysis of γδ and αβ T cells transduced with a second-generation PSCA-targeting CAR (PSCA-8t28z). We found that although γδ and αβ CAR-T cells exhibit comparable levels of cytotoxicity, they differ phenotypically. Through a system-level phosphoproteomic analysis, we identified 307 phosphosites whose abundance differed between γδ and αβ CAR-T cells. Pathway enrichment analysis placed glycolysis/gluconeogenesis and TCR signaling within the top significantly overrepresented signaling networks. The results of functional validation studies confirmed that γδ CAR-T cells have lower glycolytic and oxidative phosphorylation capacity than αβ-CAR-T cells do and weaker activation of activator protein 1 (AP-1). Notably, we identified thioredoxin-interacting protein (TXNIP) as a potential actionable target to enhance γδ CAR-T-cell metabolism. Finally, we designed a new synthetic costimulatory receptor that potentiates AP-1 activation, resulting in improved in vivo persistence. These results highlight the fundamental biological differences between γδ and αβ T cells and support the development of cell type-specific receptor engineering strategies to maximize γδ CAR-T-cell function and therapeutic benefit.
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BibTeX
@article{bustos2026signaling,
title = {{CAR signaling instructs divergent metabolic reprogramming and functional fates in αβ and γδ T cells}},
author = {Xiomar E. Bustos and Leticia Tordesillas and Elena Martinez Planes and Miguel G. Fontela and Sebastian Snedal and Renata Ariza Marques Rossetti and Victoria Izumi and Bin Fang and John M. Koomen and Eric A. Welsh and Patrick Hwu and Daniel Abate-Daga},
journal = {Cellular and Molecular Immunology},
year = {2026},
month = sep,
doi = {10.1038/s41423-026-01475-y},
url = {https://doi.org/10.1038/s41423-026-01475-y},
note = {CC BY 4.0},
}
RIS
TY - JOUR
TI - CAR signaling instructs divergent metabolic reprogramming and functional fates in αβ and γδ T cells
AU - Xiomar E. Bustos
AU - Leticia Tordesillas
AU - Elena Martinez Planes
AU - Miguel G. Fontela
AU - Sebastian Snedal
AU - Renata Ariza Marques Rossetti
AU - Victoria Izumi
AU - Bin Fang
AU - John M. Koomen
AU - Eric A. Welsh
AU - Patrick Hwu
AU - Daniel Abate-Daga
PY - 2026
DA - 2026/09/30
JO - Cellular and Molecular Immunology
DO - 10.1038/s41423-026-01475-y
UR - https://doi.org/10.1038/s41423-026-01475-y
AB - γδ T-cell-based immunotherapies have become relevant as alternatives to conventional αβ T-cell products, with preclinical data demonstrating tumor burden reduction and the mitigation of tumor-induced tissue damage. Given that most CAR constructs have been optimized for αβ T cells, we hypothesized that distinct T-cell types may require tailored CAR architectures to achieve optimal function. To test this hypothesis, we conducted a systematic comparative analysis of γδ and αβ T cells transduced with a second-generation PSCA-targeting CAR (PSCA-8t28z). We found that although γδ and αβ CAR-T cells exhibit comparable levels of cytotoxicity, they differ phenotypically. Through a system-level phosphoproteomic analysis, we identified 307 phosphosites whose abundance differed between γδ and αβ CAR-T cells. Pathway enrichment analysis placed glycolysis/gluconeogenesis and TCR signaling within the top significantly overrepresented signaling networks. The results of functional validation studies confirmed that γδ CAR-T cells have lower glycolytic and oxidative phosphorylation capacity than αβ-CAR-T cells do and weaker activation of activator protein 1 (AP-1). Notably, we identified thioredoxin-interacting protein (TXNIP) as a potential actionable target to enhance γδ CAR-T-cell metabolism. Finally, we designed a new synthetic costimulatory receptor that potentiates AP-1 activation, resulting in improved in vivo persistence. These results highlight the fundamental biological differences between γδ and αβ T cells and support the development of cell type-specific receptor engineering strategies to maximize γδ CAR-T-cell function and therapeutic benefit.
SN - 1672-7681
SN - 2042-0226
ER -