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Updated: Jul 17, 2025

Author Spotlight: Advancements in Hypoxia-Sensitive CAR-T Therapy for Enhanced Cancer Immunotherapy
Published on: June 14, 2024
FOXO1 Inhibition Generates Potent Nonactivated CAR T Cells against Solid Tumors.
Maude Marchais1,2, Luca Simula2, Mélanie Phayanouvong3
1CNRS UMR9196, Physiologie et Pathologie Moléculaires des Rétrovirus Endogènes et Infectieux, Gustave Roussy, Faculté de Médecine, Université Paris-Sud, Université Paris-Saclay, Villejuif, France.
Researchers developed a new method to create chimeric antigen receptor (CAR) T cells without ex vivo preactivation by inhibiting FOXO1. This novel CAR T-cell therapy enhances antitumor functions and shows improved efficacy against solid tumors.
Area of Science:
- Immunology
- Cellular Therapy
- Oncology
Background:
- Chimeric antigen receptor (CAR) T-cell therapy shows promise for B-cell malignancies but faces manufacturing challenges.
- Ex vivo activation, while necessary for CAR T-cell expansion, can reduce their antitumor potential.
- Improving CAR T-cell properties is crucial for enhancing therapy efficacy, especially for solid tumors.
Purpose of the Study:
- To develop a protocol for generating CAR T cells without ex vivo preactivation.
- To investigate the impact of inhibiting the transcription factor FOXO1 on T-cell function and CAR expression.
- To evaluate the therapeutic potential of these modified CAR T cells (CAR TAS) against solid tumors.
Main Methods:
- Inhibition of the transcription factor FOXO1 in primary T cells to create T-cell Activation and Survival (TAS) cells.
- Lentiviral transduction of TAS cells to generate CAR TAS cells without ex vivo preactivation.
- Assessment of CAR TAS cell phenotype, function (proliferation, cytotoxicity, migration), and in vivo efficacy in solid tumor models.
Main Results:
- FOXO1 inhibition rendered T cells permissive to lentiviral infection, enabling CAR expression.
- CAR TAS cells exhibited a stem cell memory phenotype, increased granzyme B and TNFα production.
- CAR TAS cells demonstrated enhanced proliferative, cytotoxic, and migratory capabilities compared to classical CAR T cells.
- In vivo studies confirmed superior solid tumor growth control by CAR TAS cells.
- The protocol's clinical feasibility was validated using patient-derived cells.
Conclusions:
- Inhibiting FOXO1 is a viable strategy to generate potent CAR T cells without ex vivo preactivation.
- CAR TAS cells possess enhanced antitumor functions, offering a promising approach for improving CAR T-cell therapy, particularly for solid tumors.
- This protocol is feasible for clinical application, paving the way for enhanced cell-based cancer immunotherapies.

