Sustained and specific multiplexed immune checkpoint modulation in CAR T cells induced by targeted epigenome editing
Engineered T cells equipped with a chimeric antigen receptor (CAR) have shown tremendous clinical success, but tumor-mediated stimulation of T cell inhibitory receptors leads to exhaustion, hampering durable remission in patients. Mitigation of this effect via checkpoint inhibition or genome editing...
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| Format: | Article |
| Language: | English |
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Elsevier
2025-09-01
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| Series: | Molecular Therapy: Nucleic Acids |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2162253125001726 |
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| author | Maria Silvia Roman Azcona Gianni Monaco Melissa Whitehead Masako Monika Kaufmann Jamal Alzubi Toni Cathomen Claudio Mussolino |
| author_facet | Maria Silvia Roman Azcona Gianni Monaco Melissa Whitehead Masako Monika Kaufmann Jamal Alzubi Toni Cathomen Claudio Mussolino |
| author_sort | Maria Silvia Roman Azcona |
| collection | DOAJ |
| description | Engineered T cells equipped with a chimeric antigen receptor (CAR) have shown tremendous clinical success, but tumor-mediated stimulation of T cell inhibitory receptors leads to exhaustion, hampering durable remission in patients. Mitigation of this effect via checkpoint inhibition or genome editing to knockout the genes encoding for these receptors has shown promise. Yet, the side effects of these procedures require better alternatives. Targeted epigenome editing offers a potent strategy to alter gene expression without DNA modifications. Its hit-and-run mechanism enables durable, multiplexed modulation of gene expression with greater safety. Here, we describe multiplexed epigenome editing inactivation of two critical-exhaustion-related genes, PDCD1 and LAG3, both in primary human T cells and in prostate-cancer-specific CAR T cells. Epigenetically modified CAR T cells are indistinguishable from parental cells across a range of functional assays. Although the model does not fully mimic T cell exhaustion, limiting functional assessment, gene silencing remains durable across multiple divisions and repeated CAR stimulations. Furthermore, transcriptomic analysis revealed minimal off-target effects not directly attributable to the effectors used. We demonstrate that targeted epigenome editing is effective and safe for multiplexed gene inhibition and holds potential in engineering CAR T cells with enhanced and customizable features. |
| format | Article |
| id | doaj-art-a9a97b04826c47b7b1978abd4dddab59 |
| institution | Kabale University |
| issn | 2162-2531 |
| language | English |
| publishDate | 2025-09-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Molecular Therapy: Nucleic Acids |
| spelling | doaj-art-a9a97b04826c47b7b1978abd4dddab592025-08-20T03:29:13ZengElsevierMolecular Therapy: Nucleic Acids2162-25312025-09-0136310261810.1016/j.omtn.2025.102618Sustained and specific multiplexed immune checkpoint modulation in CAR T cells induced by targeted epigenome editingMaria Silvia Roman Azcona0Gianni Monaco1Melissa Whitehead2Masako Monika Kaufmann3Jamal Alzubi4Toni Cathomen5Claudio Mussolino6Institute for Transfusion Medicine and Gene Therapy, Medical Center - University of Freiburg, 79106 Freiburg, Germany; Center for Chronic Immunodeficiency (CCI), Medical Center - University of Freiburg, 79106 Freiburg, GermanyInstitute for Transfusion Medicine and Gene Therapy, Medical Center - University of Freiburg, 79106 Freiburg, Germany; Center for Chronic Immunodeficiency (CCI), Medical Center - University of Freiburg, 79106 Freiburg, GermanyInstitute for Transfusion Medicine and Gene Therapy, Medical Center - University of Freiburg, 79106 Freiburg, Germany; Center for Chronic Immunodeficiency (CCI), Medical Center - University of Freiburg, 79106 Freiburg, Germany; Ph.D. Program, Faculty of Biology, University of Freiburg, 79104 Freiburg, GermanyInstitute for Transfusion Medicine and Gene Therapy, Medical Center - University of Freiburg, 79106 Freiburg, Germany; Center for Chronic Immunodeficiency (CCI), Medical Center - University of Freiburg, 79106 Freiburg, Germany; Spemann Graduate School of Biology and Medicine (SGBM), University of Freiburg, Freiburg, GermanyInstitute for Transfusion Medicine and Gene Therapy, Medical Center - University of Freiburg, 79106 Freiburg, Germany; Center for Chronic Immunodeficiency (CCI), Medical Center - University of Freiburg, 79106 Freiburg, GermanyInstitute for Transfusion Medicine and Gene Therapy, Medical Center - University of Freiburg, 79106 Freiburg, Germany; Center for Chronic Immunodeficiency (CCI), Medical Center - University of Freiburg, 79106 Freiburg, Germany; Faculty of Medicine, University of Freiburg, 79106 Freiburg, GermanyInstitute for Transfusion Medicine and Gene Therapy, Medical Center - University of Freiburg, 79106 Freiburg, Germany; Center for Chronic Immunodeficiency (CCI), Medical Center - University of Freiburg, 79106 Freiburg, Germany; Faculty of Medicine, University of Freiburg, 79106 Freiburg, Germany; Corresponding author: Claudio Mussolino, Institute for Transfusion Medicine and Gene Therapy, Medical Center - University of Freiburg, Freiburg 79106, Germany.Engineered T cells equipped with a chimeric antigen receptor (CAR) have shown tremendous clinical success, but tumor-mediated stimulation of T cell inhibitory receptors leads to exhaustion, hampering durable remission in patients. Mitigation of this effect via checkpoint inhibition or genome editing to knockout the genes encoding for these receptors has shown promise. Yet, the side effects of these procedures require better alternatives. Targeted epigenome editing offers a potent strategy to alter gene expression without DNA modifications. Its hit-and-run mechanism enables durable, multiplexed modulation of gene expression with greater safety. Here, we describe multiplexed epigenome editing inactivation of two critical-exhaustion-related genes, PDCD1 and LAG3, both in primary human T cells and in prostate-cancer-specific CAR T cells. Epigenetically modified CAR T cells are indistinguishable from parental cells across a range of functional assays. Although the model does not fully mimic T cell exhaustion, limiting functional assessment, gene silencing remains durable across multiple divisions and repeated CAR stimulations. Furthermore, transcriptomic analysis revealed minimal off-target effects not directly attributable to the effectors used. We demonstrate that targeted epigenome editing is effective and safe for multiplexed gene inhibition and holds potential in engineering CAR T cells with enhanced and customizable features.http://www.sciencedirect.com/science/article/pii/S2162253125001726MT: Delivery Strategiesepigenome editingimmunotherapyCAR T cellsimmune checkpointsprostate cancer |
| spellingShingle | Maria Silvia Roman Azcona Gianni Monaco Melissa Whitehead Masako Monika Kaufmann Jamal Alzubi Toni Cathomen Claudio Mussolino Sustained and specific multiplexed immune checkpoint modulation in CAR T cells induced by targeted epigenome editing Molecular Therapy: Nucleic Acids MT: Delivery Strategies epigenome editing immunotherapy CAR T cells immune checkpoints prostate cancer |
| title | Sustained and specific multiplexed immune checkpoint modulation in CAR T cells induced by targeted epigenome editing |
| title_full | Sustained and specific multiplexed immune checkpoint modulation in CAR T cells induced by targeted epigenome editing |
| title_fullStr | Sustained and specific multiplexed immune checkpoint modulation in CAR T cells induced by targeted epigenome editing |
| title_full_unstemmed | Sustained and specific multiplexed immune checkpoint modulation in CAR T cells induced by targeted epigenome editing |
| title_short | Sustained and specific multiplexed immune checkpoint modulation in CAR T cells induced by targeted epigenome editing |
| title_sort | sustained and specific multiplexed immune checkpoint modulation in car t cells induced by targeted epigenome editing |
| topic | MT: Delivery Strategies epigenome editing immunotherapy CAR T cells immune checkpoints prostate cancer |
| url | http://www.sciencedirect.com/science/article/pii/S2162253125001726 |
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