A Novel High-Throughput Framework to Quantify Spatio-Temporal Tumor Clonal Dynamics

Clonal proliferation dynamics within a tumor channels the course of tumor growth, drug response and activity. A high-throughput image screening technique is required to analyze and quantify the spatiotemporal variations in cell proliferation and influence of chemotherapy on clonal colonies. We present two protocols for generating spatial, Lentiviral Gene Ontology (LeGO) fluorescent tag based, mono- and co culture systems with provisions for spatio-temporal tracking of clonal growth at the nucleus- and cytoplasm-level. The cultured cells are subjected to either drug treatment or co-cultured with fibroblasts and analyzed with a novel image processing framework. This framework enables alignment of cell positions based on motion capture techniques, tracking through time and investigation of drug actions or co-culturing on individual cell colonies. Finally, utilizing this framework, we develop agent-based models to simulate and predict the effects of the microenvironment and clonal density on cell proliferation. The model and experimental findings suggest growth stimulating effects of local clonal density irrespective of overall cell confluency.