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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...

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Autores principales: 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
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
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.
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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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