Abstract
This study introduces Behavior-Guided Transcriptomics (BGT), an extension to the BEHAV3D protocol, integrating T cell live imaging data with single-cell RNA sequencing. The method maps gene programs to specific dynamic behaviors of T cells in 3D tumor organoid cocultures. It enables prediction of transcriptional states based on imaging-defined behavior classes such as “super engagers”, providing novel biomarkers for immunotherapeutic development. The protocol includes coculture setup, behavior-driven sorting, scRNA-seq, and computational behavioral probability mapping. BGT expands the analytical toolkit for dissecting immune cell behavior and efficacy in cancer treatment.
Type
Publication
Nature Protocols