T-BET promotes inflammatory specialization without driving terminal maturation of iPSC-derived NK cells
5 affiliations
- 1Center for Hematology and Regenerative Medicine, Department of Medicine Huddinge
- 2Department of Clinical Sciences, Intervention and Technology, Karolinska Institutet
- 3Gynecology and Reproductive Medicine, Karolinska Universitetssjukhuset
- 4Lund Stem Cell Center and Division of Molecular Hematology, Department of Laboratory Medicine, Lund University
- 5Hematology Center, Karolinska Universitetssjukhuset
Abstract
Induced pluripotent stem cells (iPSCs) provide a scalable platform for off-the-shelf immune cell therapies; however, the mechanisms controlling maturation and functional specialization of iPSC-derived natural killer (iNK) cells are not fully understood. In this study, we combined single-cell transcriptomics, phenotypic profiling, and functional assays to characterize iNK cells and to determine whether temporally controlled expression of the transcription factor T-BET (TBX21) can promote iNK terminal specification and effector functions. Single-cell RNA sequencing and phenotypic profiling confirmed that baseline iNK cells form a transcriptionally distinct NK population and retain a developmental transcriptional signature. These iNK cells exhibited incomplete terminal maturation compared with peripheral blood NK cells, expressing much of the cytotoxic machinery but displaying limited acquisition of maturation-associated receptors. Despite induction of selected terminal specification markers, T-BET did not shift iNK cells toward a peripheral NK-like transcriptional identity. Instead, T-BET selectively triggered an inflammatory effector program characterized by elevated CCL3 , CCL5 and GZMA expression and increased IFNγ production across multiple tumor targets, without enhancement of terminal cytotoxic gene programs. These findings suggest a functional dissociation between inflammatory and cytotoxic programs in iPSC-derived NK cells. This work highlights transcriptional engineering as a strategy to modulate functional state in stem cell-derived NK cells.
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BibTeX
@article{marsal2026tbet,
title = {{T-BET promotes inflammatory specialization without driving terminal maturation of iPSC-derived NK cells}},
author = {Claire Marsal and Muhammad Kashif and Sophia Borate and Charlotte Hållstrand and Linda Haazen and Mari Gilljam and Yagmur Dölalan and Nutsa Burduli and Stephan Meinke and Anders Norman and Alamdar Hussain and Fredrik Lanner and Ewa Sitnicka and Evren Alici},
journal = {Frontiers in Pharmacology},
year = {2026},
month = oct,
doi = {10.3389/fphar.2026.1952429},
url = {https://www.frontiersin.org/articles/10.3389/fphar.2026.1952429/full},
note = {CC BY 4.0},
}
RIS
TY - JOUR
TI - T-BET promotes inflammatory specialization without driving terminal maturation of iPSC-derived NK cells
AU - Claire Marsal
AU - Muhammad Kashif
AU - Sophia Borate
AU - Charlotte Hållstrand
AU - Linda Haazen
AU - Mari Gilljam
AU - Yagmur Dölalan
AU - Nutsa Burduli
AU - Stephan Meinke
AU - Anders Norman
AU - Alamdar Hussain
AU - Fredrik Lanner
AU - Ewa Sitnicka
AU - Evren Alici
PY - 2026
DA - 2026/10/27
JO - Frontiers in Pharmacology
DO - 10.3389/fphar.2026.1952429
UR - https://www.frontiersin.org/articles/10.3389/fphar.2026.1952429/full
AB - Induced pluripotent stem cells (iPSCs) provide a scalable platform for off-the-shelf immune cell therapies; however, the mechanisms controlling maturation and functional specialization of iPSC-derived natural killer (iNK) cells are not fully understood. In this study, we combined single-cell transcriptomics, phenotypic profiling, and functional assays to characterize iNK cells and to determine whether temporally controlled expression of the transcription factor T-BET (TBX21) can promote iNK terminal specification and effector functions. Single-cell RNA sequencing and phenotypic profiling confirmed that baseline iNK cells form a transcriptionally distinct NK population and retain a developmental transcriptional signature. These iNK cells exhibited incomplete terminal maturation compared with peripheral blood NK cells, expressing much of the cytotoxic machinery but displaying limited acquisition of maturation-associated receptors. Despite induction of selected terminal specification markers, T-BET did not shift iNK cells toward a peripheral NK-like transcriptional identity. Instead, T-BET selectively triggered an inflammatory effector program characterized by elevated CCL3 , CCL5 and GZMA expression and increased IFNγ production across multiple tumor targets, without enhancement of terminal cytotoxic gene programs. These findings suggest a functional dissociation between inflammatory and cytotoxic programs in iPSC-derived NK cells. This work highlights transcriptional engineering as a strategy to modulate functional state in stem cell-derived NK cells.
SN - 1663-9812
ER -