PD-L1 CAR effector cells induce self-amplifying cytotoxic effects against target cells

Background Immune checkpoint inhibitors and chimeric antigen receptor (CAR)-based therapies have transformed cancer treatment. Recently, combining these approaches into a strategy of PD-L1-targeted CAR has been proposed to target PD-L1high tumors. Our study provides new information on the efficacy o...

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Main Authors: Leszek Pączek, Radoslaw Zagozdzon, Karl-Johan Malmberg, Malgorzata Bajor, Agnieszka Graczyk-Jarzynka, Katsiaryna Marhelava, Anna Burdzinska, Angelika Muchowicz, Agnieszka Goral, Andriy Zhylko, Karolina Soroczynska, Kuba Retecki, Marta Krawczyk, Marta Klopotowska, Zofia Pilch, Sébastien Wälchli, Magdalena Winiarska
Format: Article
Language:English
Published: BMJ Publishing Group 2022-01-01
Series:Journal for ImmunoTherapy of Cancer
Online Access:https://jitc.bmj.com/content/10/1/e002500.full
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author Leszek Pączek
Radoslaw Zagozdzon
Karl-Johan Malmberg
Malgorzata Bajor
Agnieszka Graczyk-Jarzynka
Katsiaryna Marhelava
Anna Burdzinska
Angelika Muchowicz
Agnieszka Goral
Andriy Zhylko
Karolina Soroczynska
Kuba Retecki
Marta Krawczyk
Marta Klopotowska
Zofia Pilch
Sébastien Wälchli
Magdalena Winiarska
author_facet Leszek Pączek
Radoslaw Zagozdzon
Karl-Johan Malmberg
Malgorzata Bajor
Agnieszka Graczyk-Jarzynka
Katsiaryna Marhelava
Anna Burdzinska
Angelika Muchowicz
Agnieszka Goral
Andriy Zhylko
Karolina Soroczynska
Kuba Retecki
Marta Krawczyk
Marta Klopotowska
Zofia Pilch
Sébastien Wälchli
Magdalena Winiarska
author_sort Leszek Pączek
collection DOAJ
description Background Immune checkpoint inhibitors and chimeric antigen receptor (CAR)-based therapies have transformed cancer treatment. Recently, combining these approaches into a strategy of PD-L1-targeted CAR has been proposed to target PD-L1high tumors. Our study provides new information on the efficacy of such an approach against PD-L1low targets.Methods New atezolizumab-based PD-L1-targeted CAR was generated and introduced into T, NK, or NK-92 cells. Breast cancer MDA-MB-231 and MCF-7 cell lines or non-malignant cells (HEK293T, HMEC, MCF-10A, or BM-MSC) were used as targets to assess the reactivity or cytotoxic activity of the PD-L1–CAR-bearing immune effector cells. Stimulation with IFNγ or with supernatants from activated CAR T cells were used to induce upregulation of PD-L1 molecule expression on the target cells. HER2–CAR T cells were used for combination with PD-L1–CAR T cells against MCF-7 cells.Results PD-L1–CAR effector cells responded vigorously with degranulation and cytokine production to PD-L1high MDA-MB-231 cells, but not to PD-L1low MCF-7 cells. However, in long-term killing assays, both MDA-MB-231 and MCF-7 cells were eliminated by the PD-L1–CAR cells, although with a delay in the case of PD-L1low MCF-7 cells. Notably, the coculture of MCF-7 cells with activated PD-L1–CAR cells led to bystander induction of PD-L1 expression on MCF-7 cells and to the unique self-amplifying effect of the PD-L1–CAR cells. Accordingly, PD-L1–CAR T cells were active not only against MDA-MD-231 and MCF-7-PD-L1 but also against MCF-7-pLVX cells in tumor xenograft models. Importantly, we have also observed potent cytotoxic effects of PD-L1–CAR cells against non-malignant MCF-10A, HMEC, and BM-MSC cells, but not against HEK293T cells that initially did not express PD-L1 and were unresponsive to the stimulation . Finally, we have observed that HER-2–CAR T cells stimulate PD-L1 expression on MCF-7 cells and therefore accelerate the functionality of PD-L1–CAR T cells when used in combination.Conclusions In summary, our studies show that CAR-effector cells trigger the expression of PD-L1 on target cells, which in case of PD-L1–CAR results in the unique self-amplification phenomenon. This self-amplifying effect could be responsible for the enhanced cytotoxicity of PD-L1–CAR T cells against both malignant and non-malignant cells and implies extensive caution in introducing PD-L1–CAR strategy into clinical studies.
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spelling doaj-art-2bc9113457e748bd8bdb6676f28d3abe2025-08-20T02:16:44ZengBMJ Publishing GroupJournal for ImmunoTherapy of Cancer2051-14262022-01-0110110.1136/jitc-2021-002500PD-L1 CAR effector cells induce self-amplifying cytotoxic effects against target cellsLeszek Pączek0Radoslaw Zagozdzon1Karl-Johan Malmberg2Malgorzata Bajor3Agnieszka Graczyk-Jarzynka4Katsiaryna Marhelava5Anna Burdzinska6Angelika Muchowicz7Agnieszka Goral8Andriy Zhylko9Karolina Soroczynska10Kuba Retecki11Marta Krawczyk12Marta Klopotowska13Zofia Pilch14Sébastien Wälchli15Magdalena Winiarska16Department of Bioinformatics, Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Warsaw, PolandLaboratory for Cellular and Genetic Therapies, Medical University of Warsaw, Warsaw, Poland2Precision Immunotherapy Alliance, Oslo, NorwayDepartment of Clinical Immunology, Medical University of Warsaw, Warszawa, Mazowieckie, PolandLaboratory of Immunology, Mossakowski Medical Research Institute, Polish Academy of Sciences, Warsaw, PolandLaboratory for Cellular and Genetic Therapies, Medical University of Warsaw, Warsaw, PolandDepartment of Immunology, Transplantology and Internal Diseases, Medical University of Warsaw, Warszawa, PolandDepartment of Immunology, Medical University of Warsaw, Warszawa, PolandDepartment of Immunology, Medical University of Warsaw, Warszawa, PolandDepartment of Immunology, Medical University of Warsaw, Warszawa, PolandDepartment of Immunology, Medical University of Warsaw, Warszawa, PolandDepartment of Immunology, Medical University of Warsaw, Warszawa, PolandLaboratory for Cellular and Genetic Therapies, Medical University of Warsaw, Warsaw, PolandDepartment of Clinical Immunology, Medical University of Warsaw, Warszawa, Mazowieckie, PolandDepartment of Immunology, Medical University of Warsaw, Warszawa, PolandTranslational Research Unit, Section of Cellular Therapy, Department of Oncology, Oslo University Hospital, Oslo, NorwayDepartment of Immunology, Medical University of Warsaw, Warszawa, PolandBackground Immune checkpoint inhibitors and chimeric antigen receptor (CAR)-based therapies have transformed cancer treatment. Recently, combining these approaches into a strategy of PD-L1-targeted CAR has been proposed to target PD-L1high tumors. Our study provides new information on the efficacy of such an approach against PD-L1low targets.Methods New atezolizumab-based PD-L1-targeted CAR was generated and introduced into T, NK, or NK-92 cells. Breast cancer MDA-MB-231 and MCF-7 cell lines or non-malignant cells (HEK293T, HMEC, MCF-10A, or BM-MSC) were used as targets to assess the reactivity or cytotoxic activity of the PD-L1–CAR-bearing immune effector cells. Stimulation with IFNγ or with supernatants from activated CAR T cells were used to induce upregulation of PD-L1 molecule expression on the target cells. HER2–CAR T cells were used for combination with PD-L1–CAR T cells against MCF-7 cells.Results PD-L1–CAR effector cells responded vigorously with degranulation and cytokine production to PD-L1high MDA-MB-231 cells, but not to PD-L1low MCF-7 cells. However, in long-term killing assays, both MDA-MB-231 and MCF-7 cells were eliminated by the PD-L1–CAR cells, although with a delay in the case of PD-L1low MCF-7 cells. Notably, the coculture of MCF-7 cells with activated PD-L1–CAR cells led to bystander induction of PD-L1 expression on MCF-7 cells and to the unique self-amplifying effect of the PD-L1–CAR cells. Accordingly, PD-L1–CAR T cells were active not only against MDA-MD-231 and MCF-7-PD-L1 but also against MCF-7-pLVX cells in tumor xenograft models. Importantly, we have also observed potent cytotoxic effects of PD-L1–CAR cells against non-malignant MCF-10A, HMEC, and BM-MSC cells, but not against HEK293T cells that initially did not express PD-L1 and were unresponsive to the stimulation . Finally, we have observed that HER-2–CAR T cells stimulate PD-L1 expression on MCF-7 cells and therefore accelerate the functionality of PD-L1–CAR T cells when used in combination.Conclusions In summary, our studies show that CAR-effector cells trigger the expression of PD-L1 on target cells, which in case of PD-L1–CAR results in the unique self-amplification phenomenon. This self-amplifying effect could be responsible for the enhanced cytotoxicity of PD-L1–CAR T cells against both malignant and non-malignant cells and implies extensive caution in introducing PD-L1–CAR strategy into clinical studies.https://jitc.bmj.com/content/10/1/e002500.full
spellingShingle Leszek Pączek
Radoslaw Zagozdzon
Karl-Johan Malmberg
Malgorzata Bajor
Agnieszka Graczyk-Jarzynka
Katsiaryna Marhelava
Anna Burdzinska
Angelika Muchowicz
Agnieszka Goral
Andriy Zhylko
Karolina Soroczynska
Kuba Retecki
Marta Krawczyk
Marta Klopotowska
Zofia Pilch
Sébastien Wälchli
Magdalena Winiarska
PD-L1 CAR effector cells induce self-amplifying cytotoxic effects against target cells
Journal for ImmunoTherapy of Cancer
title PD-L1 CAR effector cells induce self-amplifying cytotoxic effects against target cells
title_full PD-L1 CAR effector cells induce self-amplifying cytotoxic effects against target cells
title_fullStr PD-L1 CAR effector cells induce self-amplifying cytotoxic effects against target cells
title_full_unstemmed PD-L1 CAR effector cells induce self-amplifying cytotoxic effects against target cells
title_short PD-L1 CAR effector cells induce self-amplifying cytotoxic effects against target cells
title_sort pd l1 car effector cells induce self amplifying cytotoxic effects against target cells
url https://jitc.bmj.com/content/10/1/e002500.full
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