Uncovering the mode of action of engineered T cells in patient cancer organoids

Extending the success of cellular immunotherapies against blood cancers to the realm of solid tumors will require improved in vitro models that reveal therapeutic modes of action at the molecular level. Here we describe a system, called BEHAV3D, developed to study the dynamic interactions of immune cells and patient cancer organoids by means of imaging and transcriptomics. We apply BEHAV3D to live-track >150,000 engineered T cells cultured with patient-derived, solid-tumor organoids, identifying a 'super engager' behavioral cluster comprising T cells with potent serial killing capacity. Among other T cell concepts we also study cancer metabolome-sensing engineered T cells (TEGs) and detect behavior-specific gene signatures that include a group of 27 genes with no previously described T cell function that are expressed by super engager killer TEGs. We further show that type I interferon can prime resistant organoids for TEG-mediated killing. BEHAV3D is a promising tool for the characterization of behavioral-phenotypic heterogeneity of cellular immunotherapies and may support the optimization of personalized solid-tumor-targeting cell therapies. The dynamics and molecular mechanisms of T cell therapies are probed in cancer organoids.

Automated summary

This study utilizes a system called BEHAV3D to investigate the dynamic interactions between immune cells and patient cancer organoids. By live-tracking a large number of engineered T cells cultured with patient-derived solid-tumor organoids, a cluster of "super engager" T cells with potent serial killing capacity is identified. The study also examines cancer metabolome-sensing engineered T cells (TEGs) and identifies behavior-specific gene signatures in these cells. Additionally, it demonstrates the ability of type I interferon to prime resistant organoids for TEG-mediated killing.