Cited by 354CAR-T cell therapy researchImmune Cell Function and InteractionImmunotherapy and Immune Responses

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3D model for CAR‐mediated cytotoxicity using patient‐derived colorectal cancer organoids

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Schnalzger, Theresa E, de Groot, Marnix HP, Zhang, Congcong, Mosa, Mohammed H, Michels, Birgitta E, Röder, Jasmin, Darvishi, Tahmineh, Wels, Winfried S, Farin, Henner F·2019·The EMBO Journal

DOI 10.15252/embj.2018100928PMC PMC6576164

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Abstract

Immunotherapy using chimeric antigen receptor (CAR)-engineered lymphocytes has shown impressive results in leukemia. However, for solid tumors such as colorectal cancer (CRC), new preclinical models are needed that allow to test CAR-mediated cytotoxicity in a tissue-like environment. Here, we developed a platform to study CAR cell cytotoxicity against 3-dimensional (3D) patient-derived colon organoids. Luciferase-based measurement served as a quantitative read-out for target cell viability. Additionally, we set up a confocal live imaging protocol to monitor effector cell recruitment and cytolytic activity at a single organoid level. As proof of principle, we demonstrated efficient targeting in diverse organoid models using CAR-engineered NK-92 cells directed toward a ubiquitous epithelial antigen (EPCAM). Tumor antigen-specific cytotoxicity was studied with CAR-NK-92 cells targeting organoids expressing EGFRvIII, a neoantigen found in several cancers. Finally, we tested a novel CAR strategy targeting FRIZZLED receptors that show increased expression in a subgroup of CRC tumors. Here, comparative killing assays with normal organoids failed to show tumor-specific activity. Taken together, we report a sensitive in vitro platform to evaluate CAR efficacy and tumor specificity in a personalized manner.

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Self-citation
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A 3D multi-compartment assembloid to study combined immune cell infiltration and cytotoxicity
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Authors: Schnalzger, T. E., de Groot, M. H., Zhang, C., Mosa, M. H., Michels, B. E., Röder, J., Darvishi, T., Wels, W. S., & Farin, H. F.
Affiliations: University of Konstanz (DE)
German Cancer Research Center (DE)
Goethe University Frankfurt (DE)
Corresponding author: Farin, Henner F
ORCID: Schnalzger, Theresa E
Zhang, Congcong
Mosa, Mohammed H
Michels, Birgitta E
Darvishi, Tahmineh
Wels, Winfried S
Farin, Henner F
Venue: The EMBO Journal
Publisher: Springer Science and Business Media LLC
Published: 2019-04-29
Volume: 38
Issue: 12
ISSN: 0261-4189 (ISSN-L)
1460-2075
DOI: 10.15252/embj.2018100928
PMC: PMC6576164
OpenAlex: W2942656692
Cited by: 354
References: 57
Citations by year: 2026: 45 · 2025: 60 · 2024: 62 · 2023: 63 · 2022: 47 · 2021: 41 · 2020: 29 · 2019: 7
FWCI: 12.96
Type: journal article
Language: en
Open access: bronze
doi.org
Licence: www.springer.com/tdm (tdm)
www.springer.com/tdm (vor)
Locations: The EMBO Journal (publishedVersion)
PubMed Central (publishedVersion)
PubMed (publishedVersion)
Funders: Frankfurt Cancer Institute (III L 5 ‐ 519/03/03.001 ‐ (0015))
German Cancer Consortium
Topics: Health Sciences › Medicine › Oncology › CAR-T cell therapy research
Life Sciences › Immunology and Microbiology › Immunology › Immune Cell Function and Interaction
Life Sciences › Immunology and Microbiology › Immunology › Immunotherapy and Immune Responses
Keywords: chimeric antigen receptor, colorectal cancer, 3D organoids, EpCAM, EGFRvIII, Frizzled receptors, tumor antigen specificity, luciferase reporter
MeSH terms: Receptors, Chimeric Antigen, Cells, Cultured, Cytotoxicity, Immunologic, Humans, Models, Biological, Organoids, Receptors, Antigen, T-Cell, Colorectal Neoplasms, Genetic Therapy, Immunotherapy, Adoptive, Tissue Culture Techniques, Tissue Scaffolds, HEK293 Cells, Primary Cell Culture

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3D model for CAR‐mediated cytotoxicity using patient‐derived colorectal cancer organoids

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