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.
Data availability
The bulk data generated in this study have been deposited in the Gene Expression Omnibus (GEO) under accession GSE284026. The single-cell sequencing supporting the findings of this study are available on GEO and can be found with the accession GSE284026. All data is publicly available and can be accessed by researchers worldwide. All data are included in the Supplementary Information or available from the authors, as are unique reagents used in this Article. The raw numbers for charts and graphs are available in the Source Data file whenever possible. Source data are provided with this paper.
Code availability
While we did not develop any new algorithms or tools for this study, we did use a number of open source tools to analyze the data. The specific tools and versions that we used are listed in the “methods” section of the paper. We will share our analysis code upon request.
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Acknowledgements
This research was funded in part or in full by the Austrian Science Fund (FWF; Grant-DOI 10.55776/P31383 to GB, Grant-DOI 10.55776/P34368 to KBS, Grant-DOI 10.55776/PAT9292223 to TG and Grant-DOI10. 55776/T1292 to VK), the DOC fellowship from the Austrian Academy of Sciences to JK, the Bayerische Forschungsstiftung Grant BAYCELLATOR to SK and the European Research Council Fund (CoG 101124203 to SK, ADG 786462, PoC 101054365 and PoC 101189004, the last three to GB). We thank BMA Nina Posch and Dr. Friedrich Fresser (all from our institute in Innsbruck) for technical assistance. Illustrations in the figures were created with BioRender.com. For open access purposes, the author has applied a CC BY public copyright license to any accepted version of the manuscript arising from this submission.
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D.H., V.K. and G.B. conceived and designed experiments. D.H., V.K., D.R., D.S., V.L., J.K., S.P., T.S., V.W., I.H., A.F., D.S., G.D., W.P., I.S., K.S., N.T., T.G. acquired data. D.R. performed bioinformatics analysis. D.H., V.K., D.R., D.S., G.D., W.P., N.T., T.G. and G.B. analyzed and interpreted data. D.H., V.K. and G.B. wrote the manuscript. A.K., S.S., S.K., Z.T., and D.W. provided feedback. All authors read, revised and approved the final manuscript.
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SK has received honoraria from Cymab, Plectonic, TCR2 Inc., Miltenyi, Galapagos, Novartis, BMS and GSK. SK is an inventor of several patents in the field of immuno-oncology. SK received license fees from TCR2 Inc and Carina Biotech. SK received research support from TCR2 Inc., Tabby Therapeutics, Catalym GmBH, Plectonic GmBH and Arcus Bioscience for work unrelated to the manuscript. V.K., J.K., K.S., D.H. and G.B. are inventors on patents related to immunological targets in the field of immuno-oncology. D.R., I.H., D.S., V.L., S.P., T.S., Z.S., V.W., A.F., G.D., W.P., I-I. S., A.K., S.S., Z.T. and T.G. declare no competing interests.
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Humer, D., Klepsch, V., Rieder, D. et al. NR2F6 deletion revives CAR-T cell function and induces antigen-agnostic immune memory in solid tumors. Nat Commun (2026). https://doi.org/10.1038/s41467-026-69796-0
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DOI: https://doi.org/10.1038/s41467-026-69796-0
