High-content screening of cells has become a widespread approach for cellular assays due to its capacity to capture complex biological processes. However, conventional cell culture is limited with respect to cell and tissue architecture. Organoids are a unique model system for the intact and diseased intestinal epithelium. The 3D model can be used for the functional study of cancer development and, potentially, prospective therapeutic testing of drugs in patient derived tumor organoids. Here, we present a high-content microscopy based screening workflow to study organoid self-organization and growth with up to single cell resolution. After seeding of organoid fragments in a basal membrane extract, screening plates are incubated to allow for organoid formation. Subsequent treatment and incubation is followed by staining and imaging on a high-throughput microscopy platform followed by automated image analysis using open-source software. Profiling of both complete organoids and their individual architecture enables the quantitative description of population and tissue heterogeneity in the context of various perturbations. We generated four distinct colon organoid lines from mice carrying mutations of APC and KRAS in different combinations. These are profiled for differential phenotypic responses to a library of >1000 drug-like compounds. Also, this methodology is used to screen for clinically relevant differential treatment responses in patient derived tumor and normal colon organoids. Hence, based on this work we are able to analyze gene-drug interactions in early colon cancer development and drug response of patient derived colorectal cancer organoids.

Citation Format: Niklas T. Rindtorff, Johannes Betge, Jan Sauer, Thilo Miersch, Tianzuo Zhan, Florian Heigwer, Claudia Scholl, Matthias Ebert, Bernd Fischer, Michael Boutros. High-content microscopy-based screening of colorectal organoids [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 5766. doi:10.1158/1538-7445.AM2017-5766