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Effect of CRISPR/Cas9-Mediated PD-1-Disrupted Primary Human Third-Generation CAR-T Cells Targeting EGFRvIII on In Vitro Human Glioblastoma Cell Growth

Glioblastoma (GBM), which is the most common malignant brain tumor, is resistant to standard treatments. Immunotherapy might be a promising alternative for the treatment of this cancer. Chimeric antigen receptor (CAR) is an artificially modified fusion protein that can be engineered to direct the sp...

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Autores principales: Nakazawa, Tsutomu, Natsume, Atsushi, Nishimura, Fumihiko, Morimoto, Takayuki, Matsuda, Ryosuke, Nakamura, Mitsutoshi, Yamada, Shuichi, Nakagawa, Ichiro, Motoyama, Yasushi, Park, Young-Soo, Tsujimura, Takahiro, Wakabayashi, Toshihiko, Nakase, Hiroyuki
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7227242/
https://www.ncbi.nlm.nih.gov/pubmed/32316275
http://dx.doi.org/10.3390/cells9040998
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author Nakazawa, Tsutomu
Natsume, Atsushi
Nishimura, Fumihiko
Morimoto, Takayuki
Matsuda, Ryosuke
Nakamura, Mitsutoshi
Yamada, Shuichi
Nakagawa, Ichiro
Motoyama, Yasushi
Park, Young-Soo
Tsujimura, Takahiro
Wakabayashi, Toshihiko
Nakase, Hiroyuki
author_facet Nakazawa, Tsutomu
Natsume, Atsushi
Nishimura, Fumihiko
Morimoto, Takayuki
Matsuda, Ryosuke
Nakamura, Mitsutoshi
Yamada, Shuichi
Nakagawa, Ichiro
Motoyama, Yasushi
Park, Young-Soo
Tsujimura, Takahiro
Wakabayashi, Toshihiko
Nakase, Hiroyuki
author_sort Nakazawa, Tsutomu
collection PubMed
description Glioblastoma (GBM), which is the most common malignant brain tumor, is resistant to standard treatments. Immunotherapy might be a promising alternative for the treatment of this cancer. Chimeric antigen receptor (CAR) is an artificially modified fusion protein that can be engineered to direct the specificity and function of T cells against tumor antigens. However, the antitumor effects of EGFRvIII-targeting CAR-T (EvCAR-T) cells in GBM are limited. The inhibitory effect is induced by the interaction between programmed cell death protein 1 (PD-1) on activated EvCAR-T cells and its ligands on GBM cells. In the present study, PD-1-disrupted EvCAR-T cells were established using the clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9). The sgRNA/Cas9 expression vectors designed precisely disrupted the target region of PD-1 and inhibited the expression of PD-1 in EvCAR-T cells. The PD-1-disrupted EvCAR-T cells had an in vitro growth inhibitory effect on EGFRvIII-expressing GBM cells without altering the T-cell phenotype and the expression of other checkpoint receptors. In the future, the in vivo antitumor effect of this vector should be evaluated in order to determine if it could be applied clinically for improving the efficacy of EvCAR-T cell-based adoptive immunotherapy for GBM.
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spelling pubmed-72272422020-05-28 Effect of CRISPR/Cas9-Mediated PD-1-Disrupted Primary Human Third-Generation CAR-T Cells Targeting EGFRvIII on In Vitro Human Glioblastoma Cell Growth Nakazawa, Tsutomu Natsume, Atsushi Nishimura, Fumihiko Morimoto, Takayuki Matsuda, Ryosuke Nakamura, Mitsutoshi Yamada, Shuichi Nakagawa, Ichiro Motoyama, Yasushi Park, Young-Soo Tsujimura, Takahiro Wakabayashi, Toshihiko Nakase, Hiroyuki Cells Article Glioblastoma (GBM), which is the most common malignant brain tumor, is resistant to standard treatments. Immunotherapy might be a promising alternative for the treatment of this cancer. Chimeric antigen receptor (CAR) is an artificially modified fusion protein that can be engineered to direct the specificity and function of T cells against tumor antigens. However, the antitumor effects of EGFRvIII-targeting CAR-T (EvCAR-T) cells in GBM are limited. The inhibitory effect is induced by the interaction between programmed cell death protein 1 (PD-1) on activated EvCAR-T cells and its ligands on GBM cells. In the present study, PD-1-disrupted EvCAR-T cells were established using the clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9). The sgRNA/Cas9 expression vectors designed precisely disrupted the target region of PD-1 and inhibited the expression of PD-1 in EvCAR-T cells. The PD-1-disrupted EvCAR-T cells had an in vitro growth inhibitory effect on EGFRvIII-expressing GBM cells without altering the T-cell phenotype and the expression of other checkpoint receptors. In the future, the in vivo antitumor effect of this vector should be evaluated in order to determine if it could be applied clinically for improving the efficacy of EvCAR-T cell-based adoptive immunotherapy for GBM. MDPI 2020-04-16 /pmc/articles/PMC7227242/ /pubmed/32316275 http://dx.doi.org/10.3390/cells9040998 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
Nakazawa, Tsutomu
Natsume, Atsushi
Nishimura, Fumihiko
Morimoto, Takayuki
Matsuda, Ryosuke
Nakamura, Mitsutoshi
Yamada, Shuichi
Nakagawa, Ichiro
Motoyama, Yasushi
Park, Young-Soo
Tsujimura, Takahiro
Wakabayashi, Toshihiko
Nakase, Hiroyuki
Effect of CRISPR/Cas9-Mediated PD-1-Disrupted Primary Human Third-Generation CAR-T Cells Targeting EGFRvIII on In Vitro Human Glioblastoma Cell Growth
title Effect of CRISPR/Cas9-Mediated PD-1-Disrupted Primary Human Third-Generation CAR-T Cells Targeting EGFRvIII on In Vitro Human Glioblastoma Cell Growth
title_full Effect of CRISPR/Cas9-Mediated PD-1-Disrupted Primary Human Third-Generation CAR-T Cells Targeting EGFRvIII on In Vitro Human Glioblastoma Cell Growth
title_fullStr Effect of CRISPR/Cas9-Mediated PD-1-Disrupted Primary Human Third-Generation CAR-T Cells Targeting EGFRvIII on In Vitro Human Glioblastoma Cell Growth
title_full_unstemmed Effect of CRISPR/Cas9-Mediated PD-1-Disrupted Primary Human Third-Generation CAR-T Cells Targeting EGFRvIII on In Vitro Human Glioblastoma Cell Growth
title_short Effect of CRISPR/Cas9-Mediated PD-1-Disrupted Primary Human Third-Generation CAR-T Cells Targeting EGFRvIII on In Vitro Human Glioblastoma Cell Growth
title_sort effect of crispr/cas9-mediated pd-1-disrupted primary human third-generation car-t cells targeting egfrviii on in vitro human glioblastoma cell growth
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7227242/
https://www.ncbi.nlm.nih.gov/pubmed/32316275
http://dx.doi.org/10.3390/cells9040998
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