3D model for CAR‐mediated cytotoxicity using patient‐derived colorectal cancer organoids
Open accessAbstract
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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Non-tumor cells from adjacent normal tissue can be used to develop control healthy organoids to confirm tumor-specific efficacy of tested drugs.
“Non-tumor cells from adjacent normal tissue in biopsies or surgically resected samples can be used to develop control healthy organoids to confirm the tumor-specific efficacy of the tested drugs155.”
From Organoids (2022) · cites this paper as [155]
The Patient samples section says healthy colon organoids were derived from non-pathological mucosa, including tumor-adjacent tissue collected after resection. The CAR targeting of tumor neoantigens section uses healthy organoids as controls to assess tumor-specific killing. The tested treatment was CAR-mediated cytotoxicity, however, not drugs generally; the paper does not establish the claimed drug-testing application. (LLM, conf=0.88)
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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
- Venue
- The EMBO Journal
- Published
- 2019-04-29
- Volume
- 38
- Issue
- 12
- ISSN
- 0261-4189 (ISSN-L)
- 1460-2075
- 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
- 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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