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Characterization of a Novel Third-Generation Anti-CD24-CAR against Ovarian Cancer
Novel therapeutic approaches against ovarian cancer (OC) are urgently needed because of its high rate of recurrence even after extensive surgery and multi-agent chemotherapy. We aimed to develop a novel anti-CD24 chimeric antigen receptor (CAR) as an immunotherapeutic approach against OC cells and c...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2019
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6387114/ https://www.ncbi.nlm.nih.gov/pubmed/30717444 http://dx.doi.org/10.3390/ijms20030660 |
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author | Klapdor, Rüdiger Wang, Shuo Morgan, Michael Dörk, Thilo Hacker, Ulrich Hillemanns, Peter Büning, Hildegard Schambach, Axel |
author_facet | Klapdor, Rüdiger Wang, Shuo Morgan, Michael Dörk, Thilo Hacker, Ulrich Hillemanns, Peter Büning, Hildegard Schambach, Axel |
author_sort | Klapdor, Rüdiger |
collection | PubMed |
description | Novel therapeutic approaches against ovarian cancer (OC) are urgently needed because of its high rate of recurrence even after extensive surgery and multi-agent chemotherapy. We aimed to develop a novel anti-CD24 chimeric antigen receptor (CAR) as an immunotherapeutic approach against OC cells and cancer stem cells (CSC). CSC represents a subpopulation of the tumor characterized by enhanced chemoresistance as well as the increased capability of self-renewal and metastasis. We designed a codon-optimized third-generation CAR containing the highly active single chain variable fragment (scFv) “SWA11” against CD24. We equipped the human NK-cell line NK-92 with the anti-CD24 CAR and an anti-CD19 control CAR using lentiviral transduction. Engineered NK-92 cells showed high cytotoxic activity against CD24-positive OC cell lines (SKOV3, OVCAR3). This effect was restricted to CD24-expressing cells as shown after lentiviral transduction of CD24-negative cell lines (A2780, HEK-293T) with CD24 transmembrane proteins. Additionally, NK-92 cells equipped with our novel anti-CD24 CAR were highly effective against patient-derived primary ovarian cancer cells. The activation of NK cells was shown by specific IFNγ secretion upon antigen stimulation. To further reduce possible off-target effects in vivo, we applied a dual-CAR approach using an anti-CD24-CD28-41BB fusion protein linked via a 2A sequence to an anti-mesothelin-CD3ζ-CAR. The dual-CAR was simultaneously active against CD24 and mesothelin expressing cells. Our novel anti-CD24-CAR showed a highly cytotoxic effect against OC cell lines and primary OC cells and will be evaluated in future in vivo trials as a promising immunotherapeutic approach against OC. |
format | Online Article Text |
id | pubmed-6387114 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-63871142019-02-27 Characterization of a Novel Third-Generation Anti-CD24-CAR against Ovarian Cancer Klapdor, Rüdiger Wang, Shuo Morgan, Michael Dörk, Thilo Hacker, Ulrich Hillemanns, Peter Büning, Hildegard Schambach, Axel Int J Mol Sci Article Novel therapeutic approaches against ovarian cancer (OC) are urgently needed because of its high rate of recurrence even after extensive surgery and multi-agent chemotherapy. We aimed to develop a novel anti-CD24 chimeric antigen receptor (CAR) as an immunotherapeutic approach against OC cells and cancer stem cells (CSC). CSC represents a subpopulation of the tumor characterized by enhanced chemoresistance as well as the increased capability of self-renewal and metastasis. We designed a codon-optimized third-generation CAR containing the highly active single chain variable fragment (scFv) “SWA11” against CD24. We equipped the human NK-cell line NK-92 with the anti-CD24 CAR and an anti-CD19 control CAR using lentiviral transduction. Engineered NK-92 cells showed high cytotoxic activity against CD24-positive OC cell lines (SKOV3, OVCAR3). This effect was restricted to CD24-expressing cells as shown after lentiviral transduction of CD24-negative cell lines (A2780, HEK-293T) with CD24 transmembrane proteins. Additionally, NK-92 cells equipped with our novel anti-CD24 CAR were highly effective against patient-derived primary ovarian cancer cells. The activation of NK cells was shown by specific IFNγ secretion upon antigen stimulation. To further reduce possible off-target effects in vivo, we applied a dual-CAR approach using an anti-CD24-CD28-41BB fusion protein linked via a 2A sequence to an anti-mesothelin-CD3ζ-CAR. The dual-CAR was simultaneously active against CD24 and mesothelin expressing cells. Our novel anti-CD24-CAR showed a highly cytotoxic effect against OC cell lines and primary OC cells and will be evaluated in future in vivo trials as a promising immunotherapeutic approach against OC. MDPI 2019-02-03 /pmc/articles/PMC6387114/ /pubmed/30717444 http://dx.doi.org/10.3390/ijms20030660 Text en © 2019 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 Klapdor, Rüdiger Wang, Shuo Morgan, Michael Dörk, Thilo Hacker, Ulrich Hillemanns, Peter Büning, Hildegard Schambach, Axel Characterization of a Novel Third-Generation Anti-CD24-CAR against Ovarian Cancer |
title | Characterization of a Novel Third-Generation Anti-CD24-CAR against Ovarian Cancer |
title_full | Characterization of a Novel Third-Generation Anti-CD24-CAR against Ovarian Cancer |
title_fullStr | Characterization of a Novel Third-Generation Anti-CD24-CAR against Ovarian Cancer |
title_full_unstemmed | Characterization of a Novel Third-Generation Anti-CD24-CAR against Ovarian Cancer |
title_short | Characterization of a Novel Third-Generation Anti-CD24-CAR against Ovarian Cancer |
title_sort | characterization of a novel third-generation anti-cd24-car against ovarian cancer |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6387114/ https://www.ncbi.nlm.nih.gov/pubmed/30717444 http://dx.doi.org/10.3390/ijms20030660 |
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