Novel co-culture strategies of tumor organoids with autologous T-cells reveal clinically relevant combinations of immune-checkpoint and targeted therapies

Abstract

Patient-derived tumor organoids (PDTOs) have become relevant pre-clinical models for therapeutic modeling since they highly recapitulate patients' responses to treatment. Nevertheless, their value for immunotherapy modeling has not been fully explored. We developed a tumor processing protocol that enables the establishment of PDTOs and tumor-infiltrating lymphocytes (TILs) isolation. By the optimization of functional assays, we compared the T-cells effector functions of matching PBMCs and TILs, demonstrating that PBMCs after co-culture and TILs after initial expansion display similar responses. In addition, the evaluation of cytokine production by fluorospot in combination with an image-based killing assay enables the screening of anti-PD-1 combinations with alternative immune checkpoint inhibitors as well as its combination with target inhibitors. Our proof-of-concept functional assays showed the potential and versatility of PDTOs and T-cell co-culture systems for immunotherapy screening. The optimization of scalable functional assays downstream co-culture represents a significant step forward to increase the value of PDTOs as pre-clinical models for immunotherapeutic screens.

Deficiency of metabolic regulator PKM2 activates the pentose phosphate pathway and generates TCF1+ progenitor CD8+ T cells to improve checkpoint blockade

“…Our studies in mouse T cells showed a potent ability of PPP agonism to induce a TCF1 + progenitor state and combine with checkpoint blockade to yield a significant tumor control and survival benefit. We therefore tested the effects of AG1 in a human immunocompetent patient-derived tumor organoid system 51 , 52 ( Fig. 7G ).…”

“…7G ). Following standard methodologies 51 , 52 , we generated patient-derived tumor organoids (PDTOs) from NSCLC patient specimens. We rapidly expanded 53 T cells from autologous peripheral blood mononuclear cells (PBMCs) donated by the patients for two weeks, either in the presence or absence of AG1 ( Fig.…”