3D model for CAR-mediated cytotoxicity using patient-derived colorectal cancer organoids.
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.
['Schnalzger|Theresa E|TE|', 'de Groot|Marnix Hp|MH|', 'Zhang|Congcong|C|', 'Mosa|Mohammed H|MH|', 'Michels|Birgitta E|BE|', 'Röder|Jasmin|J|', 'Darvishi|Tahmineh|T|', 'Wels|Winfried S|WS|', 'Farin|Henner F|HF|0000-0003-1558-5366']
D002478:Cells, Cultured D015179:Colorectal Neoplasms D003602:Cytotoxicity, Immunologic D015316:Genetic Therapy D057809:HEK293 Cells D006801:Humans D016219:Immunotherapy, Adoptive D008954:Models, Biological D009940:Organoids D061251:Primary Cell Culture D011948:Receptors, Antigen, T-Cell D000076962:Receptors, Chimeric Antigen D046509:Tissue Culture Techniques D054457:Tissue Scaffolds
colorectal cancer CAR immunotherapy cytotoxicity assays natural killer cells patient‐derived organoids