Simultaneous Genetic Ablation of PD-1, LAG-3, and TIM-3 in CD8 T Cells Delays Tumor Growth and Improves Survival Outcome
Immune checkpoint inhibitors (ICI) represented a step forward in improving the outcome of patients with various refractory solid tumors and several therapeutic regimens incorporating ICI have already been approved for a variety of tumor entities. However, besides remarkable long-term responses, chec...
| Main Authors: | , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2022-03-01
|
| Series: | International Journal of Molecular Sciences |
| Subjects: | |
| Online Access: | https://www.mdpi.com/1422-0067/23/6/3207 |
| _version_ | 1797470802868699136 |
|---|---|
| author | Elisa Ciraolo Stefanie Althoff Josefine Ruß Stanislav Rosnev Monique Butze Miriam Pühl Marco Frentsch Lars Bullinger Il-Kang Na |
| author_facet | Elisa Ciraolo Stefanie Althoff Josefine Ruß Stanislav Rosnev Monique Butze Miriam Pühl Marco Frentsch Lars Bullinger Il-Kang Na |
| author_sort | Elisa Ciraolo |
| collection | DOAJ |
| description | Immune checkpoint inhibitors (ICI) represented a step forward in improving the outcome of patients with various refractory solid tumors and several therapeutic regimens incorporating ICI have already been approved for a variety of tumor entities. However, besides remarkable long-term responses, checkpoint inhibition can trigger severe immune-related adverse events in some patients. In order to improve safety of ICI as well as T cell therapy, we tested the feasibility of combining T cell-based immunotherapy with genetic disruption of checkpoint molecule expression. Therefore, we generated H-Y and ovalbumin antigen-specific CD8<sup>+</sup> T cells with abolished PD-1, LAG-3, and TIM-3 expression through CRISPR/Cas9 technology. CD8<sup>+</sup> T cells, subjected to PD-1, LAG-3, and TIM-3 genetic editing, showed a strong reduction in immune checkpoint molecule expression after in vitro activation, while no relevant reduction in responsiveness to in vitro stimulation was observed. At the same time, in B16-OVA tumor model, transferred genetically edited OT-1 CD8<sup>+</sup> T cells promoted longer survival compared to control T cells and showed enhanced expansion without associated toxicity. Our study supports the notion that antigen-specific adoptive T cell therapy with concomitant genetic disruption of multiple checkpoint inhibitory receptors could represent an effective antitumor immunotherapy approach with improved tolerability profile. |
| first_indexed | 2024-03-09T19:42:04Z |
| format | Article |
| id | doaj.art-1417179e7191449ba113e03a628c88d5 |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-09T19:42:04Z |
| publishDate | 2022-03-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-1417179e7191449ba113e03a628c88d52023-11-24T01:34:53ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672022-03-01236320710.3390/ijms23063207Simultaneous Genetic Ablation of PD-1, LAG-3, and TIM-3 in CD8 T Cells Delays Tumor Growth and Improves Survival OutcomeElisa Ciraolo0Stefanie Althoff1Josefine Ruß2Stanislav Rosnev3Monique Butze4Miriam Pühl5Marco Frentsch6Lars Bullinger7Il-Kang Na8Experimental and Clinical Research Center, Max Delbrück Center for Molecular Medicine and Charité Universitätsmedizin Berlin, 13125 Berlin, GermanyExperimental and Clinical Research Center, Max Delbrück Center for Molecular Medicine and Charité Universitätsmedizin Berlin, 13125 Berlin, GermanyExperimental and Clinical Research Center, Max Delbrück Center for Molecular Medicine and Charité Universitätsmedizin Berlin, 13125 Berlin, GermanyDepartment of Hematology, Oncology and Tumor Immunology, Charité-Universitätsmedizin Berlin, Corporate Member of Freie Universität Berlin, Humboldt-Universität zu Berlin, and Berlin Institute of Health, 10117 Berlin, GermanyExperimental and Clinical Research Center, Max Delbrück Center for Molecular Medicine and Charité Universitätsmedizin Berlin, 13125 Berlin, GermanyExperimental and Clinical Research Center, Max Delbrück Center for Molecular Medicine and Charité Universitätsmedizin Berlin, 13125 Berlin, GermanyDepartment of Hematology, Oncology and Tumor Immunology, Charité-Universitätsmedizin Berlin, Corporate Member of Freie Universität Berlin, Humboldt-Universität zu Berlin, and Berlin Institute of Health, 10117 Berlin, GermanyExperimental and Clinical Research Center, Max Delbrück Center for Molecular Medicine and Charité Universitätsmedizin Berlin, 13125 Berlin, GermanyExperimental and Clinical Research Center, Max Delbrück Center for Molecular Medicine and Charité Universitätsmedizin Berlin, 13125 Berlin, GermanyImmune checkpoint inhibitors (ICI) represented a step forward in improving the outcome of patients with various refractory solid tumors and several therapeutic regimens incorporating ICI have already been approved for a variety of tumor entities. However, besides remarkable long-term responses, checkpoint inhibition can trigger severe immune-related adverse events in some patients. In order to improve safety of ICI as well as T cell therapy, we tested the feasibility of combining T cell-based immunotherapy with genetic disruption of checkpoint molecule expression. Therefore, we generated H-Y and ovalbumin antigen-specific CD8<sup>+</sup> T cells with abolished PD-1, LAG-3, and TIM-3 expression through CRISPR/Cas9 technology. CD8<sup>+</sup> T cells, subjected to PD-1, LAG-3, and TIM-3 genetic editing, showed a strong reduction in immune checkpoint molecule expression after in vitro activation, while no relevant reduction in responsiveness to in vitro stimulation was observed. At the same time, in B16-OVA tumor model, transferred genetically edited OT-1 CD8<sup>+</sup> T cells promoted longer survival compared to control T cells and showed enhanced expansion without associated toxicity. Our study supports the notion that antigen-specific adoptive T cell therapy with concomitant genetic disruption of multiple checkpoint inhibitory receptors could represent an effective antitumor immunotherapy approach with improved tolerability profile.https://www.mdpi.com/1422-0067/23/6/3207adoptive immunotherapyT lymphocytesCRISPR/Cas9checkpoint inhibitory molecule |
| spellingShingle | Elisa Ciraolo Stefanie Althoff Josefine Ruß Stanislav Rosnev Monique Butze Miriam Pühl Marco Frentsch Lars Bullinger Il-Kang Na Simultaneous Genetic Ablation of PD-1, LAG-3, and TIM-3 in CD8 T Cells Delays Tumor Growth and Improves Survival Outcome International Journal of Molecular Sciences adoptive immunotherapy T lymphocytes CRISPR/Cas9 checkpoint inhibitory molecule |
| title | Simultaneous Genetic Ablation of PD-1, LAG-3, and TIM-3 in CD8 T Cells Delays Tumor Growth and Improves Survival Outcome |
| title_full | Simultaneous Genetic Ablation of PD-1, LAG-3, and TIM-3 in CD8 T Cells Delays Tumor Growth and Improves Survival Outcome |
| title_fullStr | Simultaneous Genetic Ablation of PD-1, LAG-3, and TIM-3 in CD8 T Cells Delays Tumor Growth and Improves Survival Outcome |
| title_full_unstemmed | Simultaneous Genetic Ablation of PD-1, LAG-3, and TIM-3 in CD8 T Cells Delays Tumor Growth and Improves Survival Outcome |
| title_short | Simultaneous Genetic Ablation of PD-1, LAG-3, and TIM-3 in CD8 T Cells Delays Tumor Growth and Improves Survival Outcome |
| title_sort | simultaneous genetic ablation of pd 1 lag 3 and tim 3 in cd8 t cells delays tumor growth and improves survival outcome |
| topic | adoptive immunotherapy T lymphocytes CRISPR/Cas9 checkpoint inhibitory molecule |
| url | https://www.mdpi.com/1422-0067/23/6/3207 |
| work_keys_str_mv | AT elisaciraolo simultaneousgeneticablationofpd1lag3andtim3incd8tcellsdelaystumorgrowthandimprovessurvivaloutcome AT stefaniealthoff simultaneousgeneticablationofpd1lag3andtim3incd8tcellsdelaystumorgrowthandimprovessurvivaloutcome AT josefineruß simultaneousgeneticablationofpd1lag3andtim3incd8tcellsdelaystumorgrowthandimprovessurvivaloutcome AT stanislavrosnev simultaneousgeneticablationofpd1lag3andtim3incd8tcellsdelaystumorgrowthandimprovessurvivaloutcome AT moniquebutze simultaneousgeneticablationofpd1lag3andtim3incd8tcellsdelaystumorgrowthandimprovessurvivaloutcome AT miriampuhl simultaneousgeneticablationofpd1lag3andtim3incd8tcellsdelaystumorgrowthandimprovessurvivaloutcome AT marcofrentsch simultaneousgeneticablationofpd1lag3andtim3incd8tcellsdelaystumorgrowthandimprovessurvivaloutcome AT larsbullinger simultaneousgeneticablationofpd1lag3andtim3incd8tcellsdelaystumorgrowthandimprovessurvivaloutcome AT ilkangna simultaneousgeneticablationofpd1lag3andtim3incd8tcellsdelaystumorgrowthandimprovessurvivaloutcome |