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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...

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Autores principales: Pascoal, Susana, Salzer, Benjamin, Scheuringer, Eva, Wenninger-Weinzierl, Andrea, Sturtzel, Caterina, Holter, Wolfgang, Taschner-Mandl, Sabine, Lehner, Manfred, Distel, Martin
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7139560/
https://www.ncbi.nlm.nih.gov/pubmed/32121414
http://dx.doi.org/10.3390/cancers12030567
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author Pascoal, Susana
Salzer, Benjamin
Scheuringer, Eva
Wenninger-Weinzierl, Andrea
Sturtzel, Caterina
Holter, Wolfgang
Taschner-Mandl, Sabine
Lehner, Manfred
Distel, Martin
author_facet Pascoal, Susana
Salzer, Benjamin
Scheuringer, Eva
Wenninger-Weinzierl, Andrea
Sturtzel, Caterina
Holter, Wolfgang
Taschner-Mandl, Sabine
Lehner, Manfred
Distel, Martin
author_sort Pascoal, Susana
collection PubMed
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.
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spelling pubmed-71395602020-04-10 A Preclinical Embryonic Zebrafish Xenograft Model to Investigate CAR T Cells in Vivo Pascoal, Susana Salzer, Benjamin Scheuringer, Eva Wenninger-Weinzierl, Andrea Sturtzel, Caterina Holter, Wolfgang Taschner-Mandl, Sabine Lehner, Manfred Distel, Martin Cancers (Basel) Article 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. MDPI 2020-02-29 /pmc/articles/PMC7139560/ /pubmed/32121414 http://dx.doi.org/10.3390/cancers12030567 Text en © 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Pascoal, Susana
Salzer, Benjamin
Scheuringer, Eva
Wenninger-Weinzierl, Andrea
Sturtzel, Caterina
Holter, Wolfgang
Taschner-Mandl, Sabine
Lehner, Manfred
Distel, Martin
A Preclinical Embryonic Zebrafish Xenograft Model to Investigate CAR T Cells in Vivo
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 Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7139560/
https://www.ncbi.nlm.nih.gov/pubmed/32121414
http://dx.doi.org/10.3390/cancers12030567
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