Dominik and Victoria et al. identified NR2F6 as a T cell-intrinsic metabolic checkpoint for CAR T cells in solid tumors. CRISPR/Cas9-mediated deletion of NR2F6 sustained a TCF1+ progenitor exhausted state and maintained metabolic fitness during chronic antigen stimulation. NR2F6 deletion in CAR T cells increased cytotoxicity, cytokine production, resistance to functional exhaustion, and tumor control in immunocompetent Panc02-EpCAM tumor models. DC-mediated cross-priming with epitope spreading and activation of endogenous immunity generated durable protection against CAR antigen-positive and -negative tumor rechallenge.
Contributed by Shishir Pant
ABSTRACT: CAR-T cell therapy is effective in hematologic malignancies but remains challenging in solid tumors owing to antigen heterogeneity and tumor microenvironment-induced exhaustion. Here, gene editing of the nuclear receptor NR2F6 restores CAR-T cell functionality, sustaining a TCF1⁺ progenitor-exhausted phenotype, enhancing metabolic fitness, and preserving cytotoxic potency under chronic antigen exposure. In immunocompetent models, Nr2f6-deficient CAR-T cells suppress solid tumor growth and induce robust, polyclonal host antitumor responses that persist after CAR-T clearance, as demonstrated by tumor re-challenge protection. Although infused CAR-T cells disappear within 2 weeks, durable tumor control coincides with epitope spreading and secondary immune responses, likely via dendritic cell reactivation. Protection against antigen-negative tumors and transferable immunity reveal a dual mode of direct cytotoxicity followed by durable immune reprogramming. This broadened host immunity may offset immune escape driven by antigen heterogeneity or loss, establishing NR2F6 inhibition as a promising CAR-T engineering strategy for durable, antigen-agnostic solid-tumor immunotherapy.
Author Info: (1) Institute for Cell Genetics, Medical University of Innsbruck, Innsbruck, Austria. (2) Institute for Cell Genetics, Medical University of Innsbruck, Innsbruck, Austria. victoria
Author Info: (1) Institute for Cell Genetics, Medical University of Innsbruck, Innsbruck, Austria. (2) Institute for Cell Genetics, Medical University of Innsbruck, Innsbruck, Austria. victoria.klepsch@i-med.ac.at. (3) Biocenter, Institute of Bioinformatics, Medical University of Innsbruck, Innsbruck, Austria. (4) Institute for Cell Genetics, Medical University of Innsbruck, Innsbruck, Austria. (5) Institute for Cell Genetics, Medical University of Innsbruck, Innsbruck, Austria. (6) Institute for Cell Genetics, Medical University of Innsbruck, Innsbruck, Austria. (7) Institute for Cell Genetics, Medical University of Innsbruck, Innsbruck, Austria. (8) Institute for Cell Genetics, Medical University of Innsbruck, Innsbruck, Austria. Department of Internal Medicine V, Haematology & Oncology, Comprehensive Cancer Center Innsbruck and Tyrolean Cancer Research Institute, Medical University of Innsbruck, Innsbruck, Austria. (9) Institute for Cell Genetics, Medical University of Innsbruck, Innsbruck, Austria. (10) Institute for Cell Genetics, Medical University of Innsbruck, Innsbruck, Austria. (11) Institute for Cell Genetics, Medical University of Innsbruck, Innsbruck, Austria. Institute of Molecular Immunology, School of Medicine and Health, Technical University of Munich, Munich, Germany. (12) Tyrolean Cancer Research Institute, Innsbruck, Austria. Department of Therapeutic Radiology and Oncology, Medical University Innsbruck, Innsbruck, Austria. (13) Institute of Hygiene and Medical Microbiology Medical University of Innsbruck, Innsbruck, Austria. (14) Institute of Hygiene and Medical Microbiology Medical University of Innsbruck, Innsbruck, Austria. (15) Tyrolean Cancer Research Institute, Innsbruck, Austria. Department of Therapeutic Radiology and Oncology, Medical University Innsbruck, Innsbruck, Austria. (16) Biocenter, Institute of Bioinformatics, Medical University of Innsbruck, Innsbruck, Austria. (17) Department of Internal Medicine V, Haematology & Oncology, Comprehensive Cancer Center Innsbruck and Tyrolean Cancer Research Institute, Medical University of Innsbruck, Innsbruck, Austria. (18) Institute for Clinical Pharmacology, Klinikum der Universitt Mnchen, Munich, Germany. German Cancer Consortium, a partnership between LMU Hospital and the DKFZ, Munich, Germany. (19) Biocenter, Institute of Bioinformatics, Medical University of Innsbruck, Innsbruck, Austria. (20) Institute for Cell Genetics, Medical University of Innsbruck, Innsbruck, Austria. (21) Institute for Cell Genetics, Medical University of Innsbruck, Innsbruck, Austria. (22) Institute for Cell Genetics, Medical University of Innsbruck, Innsbruck, Austria. (23) Department of Internal Medicine V, Haematology & Oncology, Comprehensive Cancer Center Innsbruck and Tyrolean Cancer Research Institute, Medical University of Innsbruck, Innsbruck, Austria. (24) Institute for Cell Genetics, Medical University of Innsbruck, Innsbruck, Austria. gottfried.baier@i-med.ac.at.
