DNA-demethylation by DAC induces MAGE expression and MAGE-specific T cell reactivity against tumors but also healthy cell subsets

Cancer testis antigens (CTAs) can be expressed in tumors, whereas expression is silenced in normal tissue except for the immune-privileged testis. This quasi-tumor-restricted expression makes CTAs attractive targets for T cell receptor (TCR) gene therapy. However, CTA-specific TCR gene therapy is on...

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Main Authors: Marije A.J. de Rooij, Miranda H. Meeuwsen, Anne K. Wouters, Dennis F.G. Remst, Renate S. Hagedoorn, Dirk M. van der Steen, Els M.E. Verdegaal, Tassilo L.A. Wachsmann, J.H. Frederik Falkenburg, Mirjam H.M. Heemskerk
Format: Article
Language:English
Published: Elsevier 2025-09-01
Series:Molecular Therapy: Oncology
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Online Access:http://www.sciencedirect.com/science/article/pii/S2950329925000876
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author Marije A.J. de Rooij
Miranda H. Meeuwsen
Anne K. Wouters
Dennis F.G. Remst
Renate S. Hagedoorn
Dirk M. van der Steen
Els M.E. Verdegaal
Tassilo L.A. Wachsmann
J.H. Frederik Falkenburg
Mirjam H.M. Heemskerk
author_facet Marije A.J. de Rooij
Miranda H. Meeuwsen
Anne K. Wouters
Dennis F.G. Remst
Renate S. Hagedoorn
Dirk M. van der Steen
Els M.E. Verdegaal
Tassilo L.A. Wachsmann
J.H. Frederik Falkenburg
Mirjam H.M. Heemskerk
author_sort Marije A.J. de Rooij
collection DOAJ
description Cancer testis antigens (CTAs) can be expressed in tumors, whereas expression is silenced in normal tissue except for the immune-privileged testis. This quasi-tumor-restricted expression makes CTAs attractive targets for T cell receptor (TCR) gene therapy. However, CTA-specific TCR gene therapy is only applicable for tumors with substantial and homogeneous CTA expression. To increase the number of patients eligible for CTA-specific TCR gene therapy, CTA expression can be upregulated with DNA-demethylating agents like 5-aza-2′-deoxycytidine (DAC). Here, we studied the effect of DAC on the recognition of a wide range of tumor cells by TCR-engineered T cells specific for the CTAs MAGE-A1, MAGE-A3/A6, or MAGE-A9. DAC treatment strongly increased MAGE expression in most tumor cell lines tested and strongly induced or improved recognition by MAGE-specific TCR-engineered T cells. However, MAGE upregulation was not limited to tumor cells but also occurred in healthy cells, resulting in MAGE-specific T cell reactivity against proliferating T and B cells. Overall, these results underscore the potential of DAC treatment to induce MAGE expression in tumor cells and to increase their sensitivity for MAGE-specific T cell therapy. However, DAC treatment can potentially result in on-target off-tumor reactivity, warranting careful consideration when using DAC as sensitizing strategy prior to adoptive transfer of CTA-specific T cells.
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spelling doaj-art-898a1ed0dba945e2b07f3b91a668f31c2025-08-20T03:58:18ZengElsevierMolecular Therapy: Oncology2950-32992025-09-0133320101810.1016/j.omton.2025.201018DNA-demethylation by DAC induces MAGE expression and MAGE-specific T cell reactivity against tumors but also healthy cell subsetsMarije A.J. de Rooij0Miranda H. Meeuwsen1Anne K. Wouters2Dennis F.G. Remst3Renate S. Hagedoorn4Dirk M. van der Steen5Els M.E. Verdegaal6Tassilo L.A. Wachsmann7J.H. Frederik Falkenburg8Mirjam H.M. Heemskerk9Department of Hematology, Leiden University Medical Center, Leiden 2333ZA, the NetherlandsDepartment of Hematology, Leiden University Medical Center, Leiden 2333ZA, the NetherlandsDepartment of Hematology, Leiden University Medical Center, Leiden 2333ZA, the NetherlandsDepartment of Hematology, Leiden University Medical Center, Leiden 2333ZA, the NetherlandsDepartment of Hematology, Leiden University Medical Center, Leiden 2333ZA, the NetherlandsDepartment of Hematology, Leiden University Medical Center, Leiden 2333ZA, the NetherlandsDepartment of Medical Oncology, Oncode Institute, Leiden University Medical Center, Leiden 2333ZA, the NetherlandsDepartment of Hematology, Leiden University Medical Center, Leiden 2333ZA, the NetherlandsDepartment of Hematology, Leiden University Medical Center, Leiden 2333ZA, the NetherlandsDepartment of Hematology, Leiden University Medical Center, Leiden 2333ZA, the Netherlands; Corresponding author: Mirjam H.M. Heemskerk, Department of Hematology, Leiden University Medical Center, 2333ZA Leiden, the Netherlands.Cancer testis antigens (CTAs) can be expressed in tumors, whereas expression is silenced in normal tissue except for the immune-privileged testis. This quasi-tumor-restricted expression makes CTAs attractive targets for T cell receptor (TCR) gene therapy. However, CTA-specific TCR gene therapy is only applicable for tumors with substantial and homogeneous CTA expression. To increase the number of patients eligible for CTA-specific TCR gene therapy, CTA expression can be upregulated with DNA-demethylating agents like 5-aza-2′-deoxycytidine (DAC). Here, we studied the effect of DAC on the recognition of a wide range of tumor cells by TCR-engineered T cells specific for the CTAs MAGE-A1, MAGE-A3/A6, or MAGE-A9. DAC treatment strongly increased MAGE expression in most tumor cell lines tested and strongly induced or improved recognition by MAGE-specific TCR-engineered T cells. However, MAGE upregulation was not limited to tumor cells but also occurred in healthy cells, resulting in MAGE-specific T cell reactivity against proliferating T and B cells. Overall, these results underscore the potential of DAC treatment to induce MAGE expression in tumor cells and to increase their sensitivity for MAGE-specific T cell therapy. However, DAC treatment can potentially result in on-target off-tumor reactivity, warranting careful consideration when using DAC as sensitizing strategy prior to adoptive transfer of CTA-specific T cells.http://www.sciencedirect.com/science/article/pii/S2950329925000876MT: Regular IssueCD8+ T cellTCRDACMAGEDNA-demethylation
spellingShingle Marije A.J. de Rooij
Miranda H. Meeuwsen
Anne K. Wouters
Dennis F.G. Remst
Renate S. Hagedoorn
Dirk M. van der Steen
Els M.E. Verdegaal
Tassilo L.A. Wachsmann
J.H. Frederik Falkenburg
Mirjam H.M. Heemskerk
DNA-demethylation by DAC induces MAGE expression and MAGE-specific T cell reactivity against tumors but also healthy cell subsets
Molecular Therapy: Oncology
MT: Regular Issue
CD8+ T cell
TCR
DAC
MAGE
DNA-demethylation
title DNA-demethylation by DAC induces MAGE expression and MAGE-specific T cell reactivity against tumors but also healthy cell subsets
title_full DNA-demethylation by DAC induces MAGE expression and MAGE-specific T cell reactivity against tumors but also healthy cell subsets
title_fullStr DNA-demethylation by DAC induces MAGE expression and MAGE-specific T cell reactivity against tumors but also healthy cell subsets
title_full_unstemmed DNA-demethylation by DAC induces MAGE expression and MAGE-specific T cell reactivity against tumors but also healthy cell subsets
title_short DNA-demethylation by DAC induces MAGE expression and MAGE-specific T cell reactivity against tumors but also healthy cell subsets
title_sort dna demethylation by dac induces mage expression and mage specific t cell reactivity against tumors but also healthy cell subsets
topic MT: Regular Issue
CD8+ T cell
TCR
DAC
MAGE
DNA-demethylation
url http://www.sciencedirect.com/science/article/pii/S2950329925000876
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