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Preclinical Evaluation of CRISPR-Edited CAR-NK-92 Cells for Off-the-Shelf Treatment of AML and B-ALL
Acute myeloid leukemia (AML) and B-cell acute lymphocytic leukemia (B-ALL) are severe blood malignancies affecting both adults and children. Chimeric antigen receptor (CAR)-based immunotherapies have proven highly efficacious in the treatment of leukemia. However, the challenge of the immune escape...
Autores principales: | , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9655234/ https://www.ncbi.nlm.nih.gov/pubmed/36361619 http://dx.doi.org/10.3390/ijms232112828 |
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author | Ureña-Bailén, Guillermo Dobrowolski, Jérôme-Maurice Hou, Yujuan Dirlam, Alicia Roig-Merino, Alicia Schleicher, Sabine Atar, Daniel Seitz, Christian Feucht, Judith Antony, Justin S. Mohammadian Gol, Tahereh Handgretinger, Rupert Mezger, Markus |
author_facet | Ureña-Bailén, Guillermo Dobrowolski, Jérôme-Maurice Hou, Yujuan Dirlam, Alicia Roig-Merino, Alicia Schleicher, Sabine Atar, Daniel Seitz, Christian Feucht, Judith Antony, Justin S. Mohammadian Gol, Tahereh Handgretinger, Rupert Mezger, Markus |
author_sort | Ureña-Bailén, Guillermo |
collection | PubMed |
description | Acute myeloid leukemia (AML) and B-cell acute lymphocytic leukemia (B-ALL) are severe blood malignancies affecting both adults and children. Chimeric antigen receptor (CAR)-based immunotherapies have proven highly efficacious in the treatment of leukemia. However, the challenge of the immune escape of cancer cells remains. The development of more affordable and ready-to-use therapies is essential in view of the costly and time-consuming preparation of primary cell-based treatments. In order to promote the antitumor function against AML and B-ALL, we transduced NK-92 cells with CD276-CAR or CD19-CAR constructs. We also attempted to enhance cytotoxicity by a gene knockout of three different inhibitory checkpoints in NK cell function (CBLB, NKG2A, TIGIT) with CRISPR-Cas9 technology. The antileukemic activity of the generated cell lines was tested with calcein and luciferase-based cytotoxicity assays in various leukemia cell lines. Both CAR-NK-92 exhibited targeted cytotoxicity and a significant boost in antileukemic function in comparison to parental NK-92. CRISPR-Cas9 knock-outs did not improve B-ALL cytotoxicity. However, triple knock-out CD276-CAR-NK-92 cells, as well as CBLB or TIGIT knock-out NK-92 cells, showed significantly enhanced cytotoxicity against U-937 or U-937 CD19/tag AML cell lines. These results indicate that the CD19-CAR and CD276-CAR-NK-92 cell lines’ cytotoxic performance is suitable for leukemia killing, making them promising off-the-shelf therapeutic candidates. The knock-out of CBLB and TIGIT in NK-92 and CD276-CAR-NK-92 should be further investigated for the treatment of AML. |
format | Online Article Text |
id | pubmed-9655234 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-96552342022-11-15 Preclinical Evaluation of CRISPR-Edited CAR-NK-92 Cells for Off-the-Shelf Treatment of AML and B-ALL Ureña-Bailén, Guillermo Dobrowolski, Jérôme-Maurice Hou, Yujuan Dirlam, Alicia Roig-Merino, Alicia Schleicher, Sabine Atar, Daniel Seitz, Christian Feucht, Judith Antony, Justin S. Mohammadian Gol, Tahereh Handgretinger, Rupert Mezger, Markus Int J Mol Sci Article Acute myeloid leukemia (AML) and B-cell acute lymphocytic leukemia (B-ALL) are severe blood malignancies affecting both adults and children. Chimeric antigen receptor (CAR)-based immunotherapies have proven highly efficacious in the treatment of leukemia. However, the challenge of the immune escape of cancer cells remains. The development of more affordable and ready-to-use therapies is essential in view of the costly and time-consuming preparation of primary cell-based treatments. In order to promote the antitumor function against AML and B-ALL, we transduced NK-92 cells with CD276-CAR or CD19-CAR constructs. We also attempted to enhance cytotoxicity by a gene knockout of three different inhibitory checkpoints in NK cell function (CBLB, NKG2A, TIGIT) with CRISPR-Cas9 technology. The antileukemic activity of the generated cell lines was tested with calcein and luciferase-based cytotoxicity assays in various leukemia cell lines. Both CAR-NK-92 exhibited targeted cytotoxicity and a significant boost in antileukemic function in comparison to parental NK-92. CRISPR-Cas9 knock-outs did not improve B-ALL cytotoxicity. However, triple knock-out CD276-CAR-NK-92 cells, as well as CBLB or TIGIT knock-out NK-92 cells, showed significantly enhanced cytotoxicity against U-937 or U-937 CD19/tag AML cell lines. These results indicate that the CD19-CAR and CD276-CAR-NK-92 cell lines’ cytotoxic performance is suitable for leukemia killing, making them promising off-the-shelf therapeutic candidates. The knock-out of CBLB and TIGIT in NK-92 and CD276-CAR-NK-92 should be further investigated for the treatment of AML. MDPI 2022-10-24 /pmc/articles/PMC9655234/ /pubmed/36361619 http://dx.doi.org/10.3390/ijms232112828 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/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 (https://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Ureña-Bailén, Guillermo Dobrowolski, Jérôme-Maurice Hou, Yujuan Dirlam, Alicia Roig-Merino, Alicia Schleicher, Sabine Atar, Daniel Seitz, Christian Feucht, Judith Antony, Justin S. Mohammadian Gol, Tahereh Handgretinger, Rupert Mezger, Markus Preclinical Evaluation of CRISPR-Edited CAR-NK-92 Cells for Off-the-Shelf Treatment of AML and B-ALL |
title | Preclinical Evaluation of CRISPR-Edited CAR-NK-92 Cells for Off-the-Shelf Treatment of AML and B-ALL |
title_full | Preclinical Evaluation of CRISPR-Edited CAR-NK-92 Cells for Off-the-Shelf Treatment of AML and B-ALL |
title_fullStr | Preclinical Evaluation of CRISPR-Edited CAR-NK-92 Cells for Off-the-Shelf Treatment of AML and B-ALL |
title_full_unstemmed | Preclinical Evaluation of CRISPR-Edited CAR-NK-92 Cells for Off-the-Shelf Treatment of AML and B-ALL |
title_short | Preclinical Evaluation of CRISPR-Edited CAR-NK-92 Cells for Off-the-Shelf Treatment of AML and B-ALL |
title_sort | preclinical evaluation of crispr-edited car-nk-92 cells for off-the-shelf treatment of aml and b-all |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9655234/ https://www.ncbi.nlm.nih.gov/pubmed/36361619 http://dx.doi.org/10.3390/ijms232112828 |
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