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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...

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Autores principales: Klapdor, Rüdiger, Wang, Shuo, Morgan, Michael, Dörk, Thilo, Hacker, Ulrich, Hillemanns, Peter, Büning, Hildegard, Schambach, Axel
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2019
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.
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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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