Rational Design and Organoid‐Based Evaluation of a Cocktail CAR‐γδ T Cell Therapy for Heterogeneous Glioblastoma

Abstract Various challenges, including tumor heterogeneity and inadequate T cell infiltration, impede the progress of chimeric antigen receptor T cell (CAR‐T) therapy for glioblastoma (GBM). To address these obstacles, a multiple step strategy is designed. Initially, literature review and bioinformatics analysis to screen a set of antigens that are heterogeneously expressed in GBM, which are designated as the target‐bank, are leveraged. Then, according to the multiplex immunohistochemistry results of each patient's tumor sample, a personalized panel of antigens based on the principle that most cancer cells in tumor tissues can be covered from the target‐bank is selected. To target these antigens, Vδ1 T cells are chosen as CAR vehicles because of its high tissue infiltration and off‐the‐shelf properties, and an optimized protocol for engineering CAR‐Vδ1 T cells with high purity and cytotoxicity, low exhaustion, and cytokine release is developed. Next, the specific panel of cocktail CAR‐Vδ1 T cells in the GBM organoids that are directly derived from the same patient's tumor is tested. The term “ prof ” cocktail therapy is coined to describe the approach using precise and rational combination of tumor antigens, organoid‐based evaluation, and fitness of Vδ1 T cells. It may accelerate development of effective CAR‐T drugs for heterogeneous solid tumors.