A Preclinical Embryonic Zebrafish Xenograft Model to Investigate CAR T Cells In Vivo
Chimeric antigen receptor (CAR) T cells have proven to be a powerful cellular therapy for B cell malignancies. Massive efforts are now being undertaken to reproduce the high efficacy of CAR T cells in the treatment of other malignancies. Here, predictive preclinical model systems are important, and...
| Main Authors: | , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2020-02-01
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| Series: | Cancers |
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| Online Access: | https://www.mdpi.com/2072-6694/12/3/567 |
| _version_ | 1797766488392728576 |
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| author | Susana Pascoal Benjamin Salzer Eva Scheuringer Andrea Wenninger-Weinzierl Caterina Sturtzel Wolfgang Holter Sabine Taschner-Mandl Manfred Lehner Martin Distel |
| author_facet | Susana Pascoal Benjamin Salzer Eva Scheuringer Andrea Wenninger-Weinzierl Caterina Sturtzel Wolfgang Holter Sabine Taschner-Mandl Manfred Lehner Martin Distel |
| author_sort | Susana Pascoal |
| collection | DOAJ |
| description | Chimeric antigen receptor (CAR) T cells have proven to be a powerful cellular therapy for B cell malignancies. Massive efforts are now being undertaken to reproduce the high efficacy of CAR T cells in the treatment of other malignancies. Here, predictive preclinical model systems are important, and the current gold standard for preclinical evaluation of CAR T cells are mouse xenografts. However, mouse xenograft assays are expensive and slow. Therefore, an additional vertebrate in vivo assay would be beneficial to bridge the gap from in vitro to mouse xenografts. Here, we present a novel assay based on embryonic zebrafish xenografts to investigate CAR T cell-mediated killing of human cancer cells. Using a CD19-specific CAR and Nalm-6 leukemia cells, we show that live observation of killing of Nalm-6 cells by CAR T cells is possible in zebrafish embryos. Furthermore, we applied Fiji macros enabling automated quantification of Nalm-6 cells and CAR T cells over time. In conclusion, we provide a proof-of-principle study that embryonic zebrafish xenografts can be used to investigate CAR T cell-mediated killing of tumor cells. This assay is cost-effective, fast, and offers live imaging possibilities to directly investigate CAR T cell migration, engagement, and killing of effector cells. |
| first_indexed | 2024-03-12T20:26:01Z |
| format | Article |
| id | doaj.art-0895cd8a570f4031ac1fafd02ede7b64 |
| institution | Directory Open Access Journal |
| issn | 2072-6694 |
| language | English |
| last_indexed | 2024-03-12T20:26:01Z |
| publishDate | 2020-02-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Cancers |
| spelling | doaj.art-0895cd8a570f4031ac1fafd02ede7b642023-08-02T00:29:26ZengMDPI AGCancers2072-66942020-02-0112356710.3390/cancers12030567cancers12030567A Preclinical Embryonic Zebrafish Xenograft Model to Investigate CAR T Cells In VivoSusana Pascoal0Benjamin Salzer1Eva Scheuringer2Andrea Wenninger-Weinzierl3Caterina Sturtzel4Wolfgang Holter5Sabine Taschner-Mandl6Manfred Lehner7Martin Distel8Innovative Cancer Models, St. Anna Children’s Cancer Research Institute, Zimmermannplatz 10, 1090 Vienna, AustriaChristian Doppler Laboratory for Next Generation CAR T Cells, Zimmermannplatz 10, 1090 Vienna, AustriaInnovative Cancer Models, St. Anna Children’s Cancer Research Institute, Zimmermannplatz 10, 1090 Vienna, AustriaInnovative Cancer Models, St. Anna Children’s Cancer Research Institute, Zimmermannplatz 10, 1090 Vienna, AustriaInnovative Cancer Models, St. Anna Children’s Cancer Research Institute, Zimmermannplatz 10, 1090 Vienna, AustriaDevelopment of Cellular Therapeutics, St. Anna Children’s Cancer Research Institute, Zimmermannplatz 10, 1090 Vienna, AustriaTumor Biology, St. Anna Children’s Cancer Research Institute, Zimmermannplatz 10, 1090 Vienna, AustriaChristian Doppler Laboratory for Next Generation CAR T Cells, Zimmermannplatz 10, 1090 Vienna, AustriaInnovative Cancer Models, St. Anna Children’s Cancer Research Institute, Zimmermannplatz 10, 1090 Vienna, AustriaChimeric antigen receptor (CAR) T cells have proven to be a powerful cellular therapy for B cell malignancies. Massive efforts are now being undertaken to reproduce the high efficacy of CAR T cells in the treatment of other malignancies. Here, predictive preclinical model systems are important, and the current gold standard for preclinical evaluation of CAR T cells are mouse xenografts. However, mouse xenograft assays are expensive and slow. Therefore, an additional vertebrate in vivo assay would be beneficial to bridge the gap from in vitro to mouse xenografts. Here, we present a novel assay based on embryonic zebrafish xenografts to investigate CAR T cell-mediated killing of human cancer cells. Using a CD19-specific CAR and Nalm-6 leukemia cells, we show that live observation of killing of Nalm-6 cells by CAR T cells is possible in zebrafish embryos. Furthermore, we applied Fiji macros enabling automated quantification of Nalm-6 cells and CAR T cells over time. In conclusion, we provide a proof-of-principle study that embryonic zebrafish xenografts can be used to investigate CAR T cell-mediated killing of tumor cells. This assay is cost-effective, fast, and offers live imaging possibilities to directly investigate CAR T cell migration, engagement, and killing of effector cells.https://www.mdpi.com/2072-6694/12/3/567zebrafish xenograftscar t cellscd19 carin vivo imaging |
| spellingShingle | Susana Pascoal Benjamin Salzer Eva Scheuringer Andrea Wenninger-Weinzierl Caterina Sturtzel Wolfgang Holter Sabine Taschner-Mandl Manfred Lehner Martin Distel A Preclinical Embryonic Zebrafish Xenograft Model to Investigate CAR T Cells In Vivo Cancers zebrafish xenografts car t cells cd19 car in vivo imaging |
| title | A Preclinical Embryonic Zebrafish Xenograft Model to Investigate CAR T Cells In Vivo |
| title_full | A Preclinical Embryonic Zebrafish Xenograft Model to Investigate CAR T Cells In Vivo |
| title_fullStr | A Preclinical Embryonic Zebrafish Xenograft Model to Investigate CAR T Cells In Vivo |
| title_full_unstemmed | A Preclinical Embryonic Zebrafish Xenograft Model to Investigate CAR T Cells In Vivo |
| title_short | A Preclinical Embryonic Zebrafish Xenograft Model to Investigate CAR T Cells In Vivo |
| title_sort | preclinical embryonic zebrafish xenograft model to investigate car t cells in vivo |
| topic | zebrafish xenografts car t cells cd19 car in vivo imaging |
| url | https://www.mdpi.com/2072-6694/12/3/567 |
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