GPX4 is a key ferroptosis regulator orchestrating T cells and CAR-T-cells sensitivity to ferroptosis

Abstract Induction of ferroptosis, an iron-dependent form of regulated cell death, holds promise as a strategy to overcome tumor resistance to conventional therapies and enhance immunotherapy responses. However, while the susceptibility of tumor cells to ferroptosis is extensively studied, limited d...

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Main Authors: Marta Kłopotowska, Iwona Baranowska, Szymon Hajduk, Anna Jurga, Natalia Leśniowska, Michał Łaźniewski, Monika Granica, Marta Krawczyk, Milena Dziewicka, Agnieszka Graczyk, Jan Słupski, Radosław Zagożdżon, Dariusz Plewczynski, Magdalena Winiarska, Malgorzata Bajor
Format: Article
Language:English
Published: Springer 2025-08-01
Series:Cancer Immunology, Immunotherapy
Subjects:
Online Access:https://doi.org/10.1007/s00262-025-04133-w
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author Marta Kłopotowska
Iwona Baranowska
Szymon Hajduk
Anna Jurga
Natalia Leśniowska
Michał Łaźniewski
Monika Granica
Marta Krawczyk
Milena Dziewicka
Agnieszka Graczyk
Jan Słupski
Radosław Zagożdżon
Dariusz Plewczynski
Magdalena Winiarska
Malgorzata Bajor
author_facet Marta Kłopotowska
Iwona Baranowska
Szymon Hajduk
Anna Jurga
Natalia Leśniowska
Michał Łaźniewski
Monika Granica
Marta Krawczyk
Milena Dziewicka
Agnieszka Graczyk
Jan Słupski
Radosław Zagożdżon
Dariusz Plewczynski
Magdalena Winiarska
Malgorzata Bajor
author_sort Marta Kłopotowska
collection DOAJ
description Abstract Induction of ferroptosis, an iron-dependent form of regulated cell death, holds promise as a strategy to overcome tumor resistance to conventional therapies and enhance immunotherapy responses. However, while the susceptibility of tumor cells to ferroptosis is extensively studied, limited data exists on the vulnerability of immune cells to disturbed iron balance and lipid peroxidation. Here, we found that T-cell stimulation rewires iron and redox homeostasis and by increasing levels of reactive oxygen species and labile iron promotes lipid peroxidation and T-cells’ ferroptosis. Upon stimulation, we detected changes in the balance of ferroptosis-suppressive proteins, including decrease of GPX4. Subsequently, we identified GPX4 as a master regulator orchestrating T/CAR-T-cells’ sensitivity to ferroptosis and observed that GPX4 inhibitors impair CAR-T cells’ antitumor functions. Our study demonstrated differential GPX4 expression and diverse susceptibility to ferroptosis between CD4⁺ and CD8⁺ T cells. Among analyzed subsets of naïve, central memory (CM), effector memory (EM), and terminally differentiated effector memory (TEMRA), CD8⁺ EM and CD8⁺ TEMRA cells exhibited the highest sensitivity to ferroptosis. We also showed that ferroptosis limited the anti-tumor efficacy of CAR-T cells, while ferroptosis inhibition improved their therapeutic effect, both in vitro and in vivo. Our findings are not only important to understand vulnerabilities of CAR-T cells but may also hold particular significance for their therapeutic development. In this context, future anticancer therapies should be carefully designed to selectively induce the ferroptosis of tumor cells without impeding cytotoxic cells’ antitumor efficacy. Additionally, we postulate that promoting less differentiated phenotype of CAR-T cells should be exploited therapeutically to create CAR-T products characterized by decreased sensitivity to ferroptosis within tumor microenvironment.
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spelling doaj-art-bc570355a65c4eeda8af14d8065c426e2025-08-20T04:03:03ZengSpringerCancer Immunology, Immunotherapy1432-08512025-08-0174911710.1007/s00262-025-04133-wGPX4 is a key ferroptosis regulator orchestrating T cells and CAR-T-cells sensitivity to ferroptosisMarta Kłopotowska0Iwona Baranowska1Szymon Hajduk2Anna Jurga3Natalia Leśniowska4Michał Łaźniewski5Monika Granica6Marta Krawczyk7Milena Dziewicka8Agnieszka Graczyk9Jan Słupski10Radosław Zagożdżon11Dariusz Plewczynski12Magdalena Winiarska13Malgorzata Bajor14Department of Immunology, Mossakowski Medical Research Institute Polish Academy of SciencesDepartment of Immunology, Mossakowski Medical Research Institute Polish Academy of SciencesDepartment of Immunology, Mossakowski Medical Research Institute Polish Academy of SciencesDepartment of Immunology, Mossakowski Medical Research Institute Polish Academy of SciencesDepartment of Immunology, Mossakowski Medical Research Institute Polish Academy of SciencesDepartment of Immunology, Medical University of WarsawDepartment of Immunology, Mossakowski Medical Research Institute Polish Academy of SciencesDepartment of Immunology, Mossakowski Medical Research Institute Polish Academy of SciencesDepartment of Immunology, Mossakowski Medical Research Institute Polish Academy of SciencesDepartment of Immunology, Mossakowski Medical Research Institute Polish Academy of SciencesDepartment of Immunology, Mossakowski Medical Research Institute Polish Academy of SciencesLaboratory of Cellular and Genetic Therapies, Medical University of WarsawCentre of New Technologies, University of WarsawDepartment of Immunology, Mossakowski Medical Research Institute Polish Academy of SciencesDepartment of Immunology, Mossakowski Medical Research Institute Polish Academy of SciencesAbstract Induction of ferroptosis, an iron-dependent form of regulated cell death, holds promise as a strategy to overcome tumor resistance to conventional therapies and enhance immunotherapy responses. However, while the susceptibility of tumor cells to ferroptosis is extensively studied, limited data exists on the vulnerability of immune cells to disturbed iron balance and lipid peroxidation. Here, we found that T-cell stimulation rewires iron and redox homeostasis and by increasing levels of reactive oxygen species and labile iron promotes lipid peroxidation and T-cells’ ferroptosis. Upon stimulation, we detected changes in the balance of ferroptosis-suppressive proteins, including decrease of GPX4. Subsequently, we identified GPX4 as a master regulator orchestrating T/CAR-T-cells’ sensitivity to ferroptosis and observed that GPX4 inhibitors impair CAR-T cells’ antitumor functions. Our study demonstrated differential GPX4 expression and diverse susceptibility to ferroptosis between CD4⁺ and CD8⁺ T cells. Among analyzed subsets of naïve, central memory (CM), effector memory (EM), and terminally differentiated effector memory (TEMRA), CD8⁺ EM and CD8⁺ TEMRA cells exhibited the highest sensitivity to ferroptosis. We also showed that ferroptosis limited the anti-tumor efficacy of CAR-T cells, while ferroptosis inhibition improved their therapeutic effect, both in vitro and in vivo. Our findings are not only important to understand vulnerabilities of CAR-T cells but may also hold particular significance for their therapeutic development. In this context, future anticancer therapies should be carefully designed to selectively induce the ferroptosis of tumor cells without impeding cytotoxic cells’ antitumor efficacy. Additionally, we postulate that promoting less differentiated phenotype of CAR-T cells should be exploited therapeutically to create CAR-T products characterized by decreased sensitivity to ferroptosis within tumor microenvironment.https://doi.org/10.1007/s00262-025-04133-wFerroptosisT cellsChimeric antigen receptorGPX4Lipid metabolismImmunotherapy
spellingShingle Marta Kłopotowska
Iwona Baranowska
Szymon Hajduk
Anna Jurga
Natalia Leśniowska
Michał Łaźniewski
Monika Granica
Marta Krawczyk
Milena Dziewicka
Agnieszka Graczyk
Jan Słupski
Radosław Zagożdżon
Dariusz Plewczynski
Magdalena Winiarska
Malgorzata Bajor
GPX4 is a key ferroptosis regulator orchestrating T cells and CAR-T-cells sensitivity to ferroptosis
Cancer Immunology, Immunotherapy
Ferroptosis
T cells
Chimeric antigen receptor
GPX4
Lipid metabolism
Immunotherapy
title GPX4 is a key ferroptosis regulator orchestrating T cells and CAR-T-cells sensitivity to ferroptosis
title_full GPX4 is a key ferroptosis regulator orchestrating T cells and CAR-T-cells sensitivity to ferroptosis
title_fullStr GPX4 is a key ferroptosis regulator orchestrating T cells and CAR-T-cells sensitivity to ferroptosis
title_full_unstemmed GPX4 is a key ferroptosis regulator orchestrating T cells and CAR-T-cells sensitivity to ferroptosis
title_short GPX4 is a key ferroptosis regulator orchestrating T cells and CAR-T-cells sensitivity to ferroptosis
title_sort gpx4 is a key ferroptosis regulator orchestrating t cells and car t cells sensitivity to ferroptosis
topic Ferroptosis
T cells
Chimeric antigen receptor
GPX4
Lipid metabolism
Immunotherapy
url https://doi.org/10.1007/s00262-025-04133-w
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