Abstract 1178: Image-based quantification of tumor-immune cell interactions in 3D cultures

Abstract

INTRODUCTION Increasing numbers of cancer patients that benefit from immunotherapies and maintain durable response requires understanding of cellular mechanisms that govern anti-tumor immune responses. However, the mechanisms of action of these treatment modalities are not fully understood and the progress in this direction is hampered by a lack of appropriate pre-clinical testing models that are both clinically relevant and suitable for routine screening of drug candidates. Therefore, we developed a robust in vitro assay that allows image based analysis of 3D cultures in a high-throughput set-up. Here, immune cells are co-cultured with cancer cells in a 3D environment which recapitulates the tumor micro-environment and its complex cellular interactions. Functional read-outs, such as active migration of immune cells towards tumoroids, infiltration of immune cells into the tumoroids and their killing lead to a better understanding of the immune-modulatory profile of different immunotherapies. MATERIAL and METHOD Tumoroids generated from cancer cell lines (e.g. breast, prostate) and colorectal cancer organoids (from HUB Organoid Technology) were cultured in protein hydrogel. Different immune cells, such as PBMCs, T cells, dendritic cells or macrophages, with and without activation were added to the 3D culture and their infiltration into tumoroids and subsequent killing was visualized using high-content microscopy. Quantification of immune cell effects was achieved with morphometric analysis with OMinerTM software. RESULTS and DISCUSSION Image-based analysis enabled the discrimination of immune-tumor cell interactions in 3D cultures. These results demonstrated the effect of immune cell targeting on the tumor progression. Different levels of immune cells infiltration and killing of tumoroids and organoids were measured depending on the activation status of immune cells. The 3D environment, both for the cell culture and image analysis, allows for measurement of spatially resolved information, not accessible by monolayer cultures or biochemical assays. CONCLUSION Our image-based platform described here allows for analysis of immunotherapy effects on different cell types that engage in a more physiologically relevant spatial setting than when culturing them in traditional 2D cultures. Visualization and quantification of these tumor-immune cells interactions offer a highly powerful tool for cancer immunotherapy drug developers to understand the mechanism of action of their treatments and ultimately translating to a better clinical performance. Citation Format: Gera Goverse, Nataliia Beztsinna, Kuan Yan, Leo Price, Lidia Daszkiewicz. Image-based quantification of tumor-immune cell interactions in 3D cultures [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1178.