Regulatory T Cell Depletion Using a CRISPR Fc-Optimized CD25 Antibody
Regulatory T cells (T<sub>regs</sub>) are major drivers behind immunosuppressive mechanisms and present a major hurdle for cancer therapy. T<sub>regs</sub> are characterized by a high expression of CD25, which is a potentially valuable target for T<sub>reg</sub> d...
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| Format: | Article |
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MDPI AG
2022-08-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/23/15/8707 |
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| author | Marit J. van Elsas Johan M. S. van der Schoot Alexander Bartels Kas Steuten Duco van Dalen Zacharias Wijfjes Carl G. Figdor Thorbald van Hall Sjoerd H. van der Burg Martijn Verdoes Ferenc A. Scheeren |
| author_facet | Marit J. van Elsas Johan M. S. van der Schoot Alexander Bartels Kas Steuten Duco van Dalen Zacharias Wijfjes Carl G. Figdor Thorbald van Hall Sjoerd H. van der Burg Martijn Verdoes Ferenc A. Scheeren |
| author_sort | Marit J. van Elsas |
| collection | DOAJ |
| description | Regulatory T cells (T<sub>regs</sub>) are major drivers behind immunosuppressive mechanisms and present a major hurdle for cancer therapy. T<sub>regs</sub> are characterized by a high expression of CD25, which is a potentially valuable target for T<sub>reg</sub> depletion to alleviate immune suppression. The preclinical anti-CD25 (αCD25) antibody, clone PC-61, has met with modest anti-tumor activity due to its capacity to clear T<sub>regs</sub> from the circulation and lymph nodes, but not those that reside in the tumor. The optimization of the Fc domain of this antibody clone has been shown to enhance the intratumoral T<sub>reg</sub> depletion capacity. Here, we generated a stable cell line that produced optimized recombinant T<sub>reg</sub>-depleting antibodies. A genome engineering strategy in which CRISPR-Cas9 was combined with homology-directed repair (CRISPR-HDR) was utilized to optimize the Fc domain of the hybridoma PC-61 for effector functions by switching it from its original rat IgG1 to a mouse IgG2a isotype. In a syngeneic tumor mouse model, the resulting αCD25-m2a (mouse IgG2a isotype) antibody mediated the effective depletion of tumor-resident T<sub>regs</sub>, leading to a high effector T cell (T<sub>eff</sub>) to T<sub>reg</sub> ratio. Moreover, a combination of αCD25-m2a and an αPD-L1 treatment augmented tumor eradication in mice, demonstrating the potential for αCD25 as a cancer immunotherapy. |
| first_indexed | 2024-03-09T10:07:41Z |
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| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-09T10:07:41Z |
| publishDate | 2022-08-01 |
| publisher | MDPI AG |
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| series | International Journal of Molecular Sciences |
| spelling | doaj.art-9b99950f014f4b1bb3ab28ac5b6c67cb2023-12-01T22:58:33ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672022-08-012315870710.3390/ijms23158707Regulatory T Cell Depletion Using a CRISPR Fc-Optimized CD25 AntibodyMarit J. van Elsas0Johan M. S. van der Schoot1Alexander Bartels2Kas Steuten3Duco van Dalen4Zacharias Wijfjes5Carl G. Figdor6Thorbald van Hall7Sjoerd H. van der Burg8Martijn Verdoes9Ferenc A. Scheeren10Department of Medical Oncology, Oncode Institute, Leiden University Medical Centre (LUMC), Albinusdreef 2, 2333 ZA Leiden, The NetherlandsDepartment of Tumor Immunology, Radboud Institute for Molecular Life Sciences, Radboud University Medical Centre, Geert Grooteplein 26, 6525 GA Nijmegen, The NetherlandsDepartment of Tumor Immunology, Radboud Institute for Molecular Life Sciences, Radboud University Medical Centre, Geert Grooteplein 26, 6525 GA Nijmegen, The NetherlandsDepartment of Tumor Immunology, Radboud Institute for Molecular Life Sciences, Radboud University Medical Centre, Geert Grooteplein 26, 6525 GA Nijmegen, The NetherlandsDepartment of Tumor Immunology, Radboud Institute for Molecular Life Sciences, Radboud University Medical Centre, Geert Grooteplein 26, 6525 GA Nijmegen, The NetherlandsDepartment of Tumor Immunology, Radboud Institute for Molecular Life Sciences, Radboud University Medical Centre, Geert Grooteplein 26, 6525 GA Nijmegen, The NetherlandsDepartment of Tumor Immunology, Radboud Institute for Molecular Life Sciences, Radboud University Medical Centre, Geert Grooteplein 26, 6525 GA Nijmegen, The NetherlandsDepartment of Medical Oncology, Oncode Institute, Leiden University Medical Centre (LUMC), Albinusdreef 2, 2333 ZA Leiden, The NetherlandsDepartment of Medical Oncology, Oncode Institute, Leiden University Medical Centre (LUMC), Albinusdreef 2, 2333 ZA Leiden, The NetherlandsDepartment of Tumor Immunology, Radboud Institute for Molecular Life Sciences, Radboud University Medical Centre, Geert Grooteplein 26, 6525 GA Nijmegen, The NetherlandsDepartment of Dermatology, Leiden University Medical Centre (LUMC), Albinusdreef 2, 2333 ZA Leiden, The NetherlandsRegulatory T cells (T<sub>regs</sub>) are major drivers behind immunosuppressive mechanisms and present a major hurdle for cancer therapy. T<sub>regs</sub> are characterized by a high expression of CD25, which is a potentially valuable target for T<sub>reg</sub> depletion to alleviate immune suppression. The preclinical anti-CD25 (αCD25) antibody, clone PC-61, has met with modest anti-tumor activity due to its capacity to clear T<sub>regs</sub> from the circulation and lymph nodes, but not those that reside in the tumor. The optimization of the Fc domain of this antibody clone has been shown to enhance the intratumoral T<sub>reg</sub> depletion capacity. Here, we generated a stable cell line that produced optimized recombinant T<sub>reg</sub>-depleting antibodies. A genome engineering strategy in which CRISPR-Cas9 was combined with homology-directed repair (CRISPR-HDR) was utilized to optimize the Fc domain of the hybridoma PC-61 for effector functions by switching it from its original rat IgG1 to a mouse IgG2a isotype. In a syngeneic tumor mouse model, the resulting αCD25-m2a (mouse IgG2a isotype) antibody mediated the effective depletion of tumor-resident T<sub>regs</sub>, leading to a high effector T cell (T<sub>eff</sub>) to T<sub>reg</sub> ratio. Moreover, a combination of αCD25-m2a and an αPD-L1 treatment augmented tumor eradication in mice, demonstrating the potential for αCD25 as a cancer immunotherapy.https://www.mdpi.com/1422-0067/23/15/8707antibodyimmunotherapyCRISPR-HDRFc optimizationhybridoma |
| spellingShingle | Marit J. van Elsas Johan M. S. van der Schoot Alexander Bartels Kas Steuten Duco van Dalen Zacharias Wijfjes Carl G. Figdor Thorbald van Hall Sjoerd H. van der Burg Martijn Verdoes Ferenc A. Scheeren Regulatory T Cell Depletion Using a CRISPR Fc-Optimized CD25 Antibody International Journal of Molecular Sciences antibody immunotherapy CRISPR-HDR Fc optimization hybridoma |
| title | Regulatory T Cell Depletion Using a CRISPR Fc-Optimized CD25 Antibody |
| title_full | Regulatory T Cell Depletion Using a CRISPR Fc-Optimized CD25 Antibody |
| title_fullStr | Regulatory T Cell Depletion Using a CRISPR Fc-Optimized CD25 Antibody |
| title_full_unstemmed | Regulatory T Cell Depletion Using a CRISPR Fc-Optimized CD25 Antibody |
| title_short | Regulatory T Cell Depletion Using a CRISPR Fc-Optimized CD25 Antibody |
| title_sort | regulatory t cell depletion using a crispr fc optimized cd25 antibody |
| topic | antibody immunotherapy CRISPR-HDR Fc optimization hybridoma |
| url | https://www.mdpi.com/1422-0067/23/15/8707 |
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