Abstract 6315: High content screening of intestinal organoid cultures to visualize and quantify effects of drug combinations

Abstract

Abstract Background: The choice in advanced pre-clinical in vitro models that use cultures of patient-derived tissues has increased but their full potential in drug discovery has not been fully exploited. Assays often lack sensitivity or clinically relevant end-points or are not suitable for high throughput screening. Here we describe a 3D screening platform that combines complex biology with high content phenotype-based image analysis to provide visualization and quantification of the effects of various targeted therapies on healthy and diseased tissue. Improved characterization of compound effects increases the scope for predicting treatment responses in patients, discriminating different drug responses and flagging off-target effects. Methods: A panel of tumor and healthy intestinal organoids, obtained from HUB Organoid Technology, were cultured in natural extracellular matrix scaffolds that support appropriate gene expression, differentiation and functional characteristics of the tissue of origin. To investigate the effects of anti-EGFR targeted therapies in combination with standard-of-care (SoC) treatments on the function, formation and integrity of colorectal cancer organoids (CRCs) and matching normal gut epithelium, we set up the following intestinal organoid test systems: 1) A panel of CRCs with a broad heterogeneity of mutations for high throughput phenotype-based screening that enable accurate compound profiling 2) Normal vs. tumor organoid pair assays for off-target effect studies and 3) Mode of action studies in various tumoroid models. Results: High content 3D image analysis of the organoids enabled sensitive detection of treatment-induced and compound-specific morphological changes beyond conventional cell viability measurements, including those associated with tumor killing, cell cycle arrest, toxicity and epithelial integrity to discriminate therapeutic and adverse responses in intestinal organoids. This enabled distinction between different mechanisms of action and determination of compound synergy. The image-based measurements can be complemented with detection of secreted factors (e.g. cytokines, chemokines) or expressed genes in response to various therapeutic compounds. Conclusion: The advanced 3D image analysis of in vitro cultured organoids described here represents a rapid and biologically relevant approach to test various anti-cancer-therapeutic modalities, including antibodies, antibody-drug conjugates and small molecules. This platform technology is suitable not only for high throughput screening in organoid panels but also in-depth mode of action studies. Moreover, evaluating effects on healthy and tumor intestinal organoids from the same patient and between patients, gives insight into potential toxicities, therapeutic window and patient-derived differences in subsequent in vivo studies. Citation Format: Mariusz Madej, Cinthya Del Angel Zuvirie, Jara García Mateos, Leo Price, Kuan Yan, Bram Herpers. High content screening of intestinal organoid cultures to visualize and quantify effects of drug combinations [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 6315.