Dekkers and Aliev et al. developed an organoid-based 3D-imaging and transcriptomic platform to live-track engineered T cells (TEGs) cultured with patient-derived organoids (PDO) and characterize the behavioral and phenotypic heterogeneity of cellular immunotherapies. TEGs exposed to PDOs showed high variation in their behavior, with “super engager” killer TEGs associated with longer tumor engagement, higher NCAM1 expression, and a specific 27-gene signature that overlapped with a cytotoxic CD8+ TIL signature from responder patients. IFN-I signaling increased the sensitivity of PDOs to TEG therapy, but didn’t impact the killing capacity of TEGs.
Contributed by Shishir Pant
ABSTRACT: 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.
Author Info: (1) Hubrecht Institute, Royal Netherlands Academy of Arts and Sciences and University Medical Center Utrecht, Utrecht, the Netherlands. Princess Mxima Center for Pediatric Oncolog
Author Info: (1) Hubrecht Institute, Royal Netherlands Academy of Arts and Sciences and University Medical Center Utrecht, Utrecht, the Netherlands. Princess Mxima Center for Pediatric Oncology, Utrecht, the Netherlands. Oncode Institute, Utrecht, the Netherlands. (2) Princess Mxima Center for Pediatric Oncology, Utrecht, the Netherlands. Oncode Institute, Utrecht, the Netherlands. (3) Center for Translational Immunology, University Medical Center Utrecht, Utrecht University, Utrecht, the Netherlands. (4) Princess Mxima Center for Pediatric Oncology, Utrecht, the Netherlands. (5) Princess Mxima Center for Pediatric Oncology, Utrecht, the Netherlands. Oncode Institute, Utrecht, the Netherlands. (6) Princess Mxima Center for Pediatric Oncology, Utrecht, the Netherlands. Oncode Institute, Utrecht, the Netherlands. (7) Hubrecht Institute, Royal Netherlands Academy of Arts and Sciences and University Medical Center Utrecht, Utrecht, the Netherlands. Oncode Institute, Utrecht, the Netherlands. Microbiome and Cancer Division, German Cancer Research Center, Heidelberg, Germany. (8) Princess Mxima Center for Pediatric Oncology, Utrecht, the Netherlands. (9) Center for Translational Immunology, University Medical Center Utrecht, Utrecht University, Utrecht, the Netherlands. (10) Center for Translational Immunology, University Medical Center Utrecht, Utrecht University, Utrecht, the Netherlands. (11) Princess Mxima Center for Pediatric Oncology, Utrecht, the Netherlands. Oncode Institute, Utrecht, the Netherlands. (12) Princess Mxima Center for Pediatric Oncology, Utrecht, the Netherlands. Oncode Institute, Utrecht, the Netherlands. (13) Princess Mxima Center for Pediatric Oncology, Utrecht, the Netherlands. Oncode Institute, Utrecht, the Netherlands. (14) Princess Mxima Center for Pediatric Oncology, Utrecht, the Netherlands. Oncode Institute, Utrecht, the Netherlands. (15) Princess Mxima Center for Pediatric Oncology, Utrecht, the Netherlands. Oncode Institute, Utrecht, the Netherlands. (16) Center for Translational Immunology, University Medical Center Utrecht, Utrecht University, Utrecht, the Netherlands. (17) Hubrecht Institute, Royal Netherlands Academy of Arts and Sciences and University Medical Center Utrecht, Utrecht, the Netherlands. Oncode Institute, Utrecht, the Netherlands. (18) Princess Mxima Center for Pediatric Oncology, Utrecht, the Netherlands. Center for Translational Immunology, University Medical Center Utrecht, Utrecht University, Utrecht, the Netherlands. (19) Hubrecht Institute, Royal Netherlands Academy of Arts and Sciences and University Medical Center Utrecht, Utrecht, the Netherlands. Oncode Institute, Utrecht, the Netherlands. (20) Hubrecht Institute, Royal Netherlands Academy of Arts and Sciences and University Medical Center Utrecht, Utrecht, the Netherlands. Oncode Institute, Utrecht, the Netherlands. (21) Center for Translational Immunology, University Medical Center Utrecht, Utrecht University, Utrecht, the Netherlands. Department of Hematology, University Medical Center Utrecht, Utrecht University, Utrecht, the Netherlands. (22) Center for Translational Immunology, University Medical Center Utrecht, Utrecht University, Utrecht, the Netherlands. (23) Center for Translational Immunology, University Medical Center Utrecht, Utrecht University, Utrecht, the Netherlands. (24) Princess Mxima Center for Pediatric Oncology, Utrecht, the Netherlands. Oncode Institute, Utrecht, the Netherlands. (25) Princess Mxima Center for Pediatric Oncology, Utrecht, the Netherlands. Oncode Institute, Utrecht, the Netherlands. (26) Princess Mxima Center for Pediatric Oncology, Utrecht, the Netherlands. Oncode Institute, Utrecht, the Netherlands. (27) Center for Translational Immunology, University Medical Center Utrecht, Utrecht University, Utrecht, the Netherlands. (28) Hubrecht Institute, Royal Netherlands Academy of Arts and Sciences and University Medical Center Utrecht, Utrecht, the Netherlands. Oncode Institute, Utrecht, the Netherlands. (29) Hubrecht Institute, Royal Netherlands Academy of Arts and Sciences and University Medical Center Utrecht, Utrecht, the Netherlands. Oncode Institute, Utrecht, the Netherlands. (30) Hubrecht Institute, Royal Netherlands Academy of Arts and Sciences and University Medical Center Utrecht, Utrecht, the Netherlands. Oncode Institute, Utrecht, the Netherlands. Mildred Scheel Early Career Center for Cancer Research Wrzburg, University Hospital Wrzburg, MSNZ/IZKF, Wurzburg, Germany. (31) Princess Mxima Center for Pediatric Oncology, Utrecht, the Netherlands. University Medical Center Utrecht, Utrecht, the Netherlands. Comprehensive Cancer Center Mnchen, Munich, Germany. (32) Princess Mxima Center for Pediatric Oncology, Utrecht, the Netherlands. Center for Translational Immunology, University Medical Center Utrecht, Utrecht University, Utrecht, the Netherlands. (33) Princess Mxima Center for Pediatric Oncology, Utrecht, the Netherlands. Oncode Institute, Utrecht, the Netherlands. (34) Princess Mxima Center for Pediatric Oncology, Utrecht, the Netherlands. (35) Hubrecht Institute, Royal Netherlands Academy of Arts and Sciences and University Medical Center Utrecht, Utrecht, the Netherlands. Princess Mxima Center for Pediatric Oncology, Utrecht, the Netherlands. Oncode Institute, Utrecht, the Netherlands. Pharma, Research and Early Development, F. Hoffmann-La Roche Ltd, Basel, Switzerland. (36) Center for Translational Immunology, University Medical Center Utrecht, Utrecht University, Utrecht, the Netherlands. Department of Hematology, University Medical Center Utrecht, Utrecht University, Utrecht, the Netherlands. (37) Center for Translational Immunology, University Medical Center Utrecht, Utrecht University, Utrecht, the Netherlands. (38) Princess Mxima Center for Pediatric Oncology, Utrecht, the Netherlands. a.c.rios@prinsesmaximacentrum.nl. Oncode Institute, Utrecht, the Netherlands. a.c.rios@prinsesmaximacentrum.nl.
