Format

Send to

Choose Destination
EMBO J. 2019 Apr 29. pii: e100928. doi: 10.15252/embj.2018100928. [Epub ahead of print]

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

Schnalzger TE1,2, de Groot MH1, Zhang C1,3,4, Mosa MH1,3,4,5, Michels BE1,3,4,5,6, Röder J1,5, Darvishi T1,3,4, Wels WS1,3,4,5, Farin HF7,3,4,5.

Author information

1
Georg-Speyer-Haus, Institute for Tumor Biology and Experimental Therapy, Frankfurt am Main, Germany.
2
University of Konstanz, Konstanz, Germany.
3
German Cancer Consortium (DKTK), Partner Site Frankfurt/Mainz, Frankfurt am Main, Germany.
4
German Cancer Research Center (DKFZ), Heidelberg, Germany.
5
Frankfurt Cancer Institute, Goethe University, Frankfurt, Germany.
6
Faculty of Biological Sciences, Goethe University, Frankfurt, Germany.
7
Georg-Speyer-Haus, Institute for Tumor Biology and Experimental Therapy, Frankfurt am Main, Germany farin@gsh.uni-frankfurt.de.

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.

KEYWORDS:

CAR immunotherapy; colorectal cancer; cytotoxicity assays; natural killer cells; patient‐derived organoids

PMID:
31036555
DOI:
10.15252/embj.2018100928

Supplemental Content

Full text links

Icon for HighWire
Loading ...
Support Center