NR2F6 functions as a key transcriptional checkpoint enforcing dysfunction of human CD3+ T cells and limiting CAR-T cell potency
Abstract
Although nuclear receptor subfamily 2 group F member 6 (NR2F6) regulates lymphocyte activation in murine models, its function in human T cells remains incompletely defined. Here, we identify NR2F6 as a T cell receptor (TCR)-inducible transcriptional checkpoint that limits effector potency in human CD3+ T cells, including chimeric antigen receptor (CAR) T cells. In an anti-EGFR CAR-T cell-A549 co-culture model, enforced NR2F6 expression, mimicking its upregulation in tumor-infiltrating lymphocytes, markedly impairs killing under repeated tumor challenge. In polyclonal T cells, NR2F6 overexpression and CRISPR-Cas9-mediated ablation produced directionally opposing effects, supporting its role as a negative regulator of activation, differentiation-associated programs, and cytokine production. Mechanistically, transcriptomic, epigenomic, and nuclear protein analyses are consistent with suppression of NFAT-, AP-1-, and NF-κB-associated pathways and reveal context-dependent changes in TCF-1-associated readouts. Collectively, these findings position NR2F6 as a signaling-integrated transcriptional checkpoint that contributes to dysfunctional T cell states and may promote tumor immune evasion in humans.
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BibTeX
@article{koutnik2026nr2f6,
title = {{NR2F6 functions as a key transcriptional checkpoint enforcing dysfunction of human CD3+ T cells and limiting CAR-T cell potency}},
author = {Jiří Koutník and Thomas Gruber and Dorothee Freimark and Dominik Humer and Daniel Schreiber and Isabel Hölzl and Viktor Lang and Nino Tokic and Tajana Sajinovic and Dominik Wolf and Nikolaus Thuille and Kerstin Siegmund and Victoria Klepsch and Gottfried Baier},
journal = {iScience},
year = {2026},
month = oct,
doi = {10.1016/j.isci.2026.117982},
url = {https://www.cell.com/iscience/fulltext/S2589-0042(26)03365-1},
note = {CC BY 4.0},
}
RIS
TY - JOUR
TI - NR2F6 functions as a key transcriptional checkpoint enforcing dysfunction of human CD3+ T cells and limiting CAR-T cell potency
AU - Jiří Koutník
AU - Thomas Gruber
AU - Dorothee Freimark
AU - Dominik Humer
AU - Daniel Schreiber
AU - Isabel Hölzl
AU - Viktor Lang
AU - Nino Tokic
AU - Tajana Sajinovic
AU - Dominik Wolf
AU - Nikolaus Thuille
AU - Kerstin Siegmund
AU - Victoria Klepsch
AU - Gottfried Baier
PY - 2026
DA - 2026/10/26
JO - iScience
DO - 10.1016/j.isci.2026.117982
UR - https://www.cell.com/iscience/fulltext/S2589-0042(26)03365-1
AB - Although nuclear receptor subfamily 2 group F member 6 (NR2F6) regulates lymphocyte activation in murine models, its function in human T cells remains incompletely defined. Here, we identify NR2F6 as a T cell receptor (TCR)-inducible transcriptional checkpoint that limits effector potency in human CD3+ T cells, including chimeric antigen receptor (CAR) T cells. In an anti-EGFR CAR-T cell-A549 co-culture model, enforced NR2F6 expression, mimicking its upregulation in tumor-infiltrating lymphocytes, markedly impairs killing under repeated tumor challenge. In polyclonal T cells, NR2F6 overexpression and CRISPR-Cas9-mediated ablation produced directionally opposing effects, supporting its role as a negative regulator of activation, differentiation-associated programs, and cytokine production. Mechanistically, transcriptomic, epigenomic, and nuclear protein analyses are consistent with suppression of NFAT-, AP-1-, and NF-κB-associated pathways and reveal context-dependent changes in TCF-1-associated readouts. Collectively, these findings position NR2F6 as a signaling-integrated transcriptional checkpoint that contributes to dysfunctional T cell states and may promote tumor immune evasion in humans.
SN - 2589-0042
ER -