GLUT1 overexpression in CAR-T cells enhances metabolic reprogramming and antitumor potency. The study shows that increasing glucose uptake via GLUT1 overexpression in CAR-T cells improves their function and antitumor effectiveness. GLUT1 overexpression increases glucose consumption, glycolysis, and oxidative phosphorylation, leading to reduced T cell exhaustion and increased Th17 differentiation. This metabolic reprogramming also enhances resistance to reactive oxygen species (ROS) and increases inosine accumulation. When challenged with tumors, GLUT1-overexpressing CAR-T cells secrete more proinflammatory cytokines, show enhanced cytotoxicity, and demonstrate superior tumor control and persistence in mouse models. The findings suggest that glucose availability is a limiting factor for CAR-T cell function, and enhancing glucose availability through GLUT1 overexpression could improve antitumor immune function. The study also shows that GLUT1 overexpression enhances metabolic pathways that favor resistance to ROS suppression, alters arginine and inosine metabolism, and improves CAR-T cell tumor clearance in vivo. Overall, the results demonstrate that GLUT1 overexpression enhances CAR-T cell potency and persistence, providing a new approach to improve the effectiveness of engineered T cell populations for antitumor targeting.GLUT1 overexpression in CAR-T cells enhances metabolic reprogramming and antitumor potency. The study shows that increasing glucose uptake via GLUT1 overexpression in CAR-T cells improves their function and antitumor effectiveness. GLUT1 overexpression increases glucose consumption, glycolysis, and oxidative phosphorylation, leading to reduced T cell exhaustion and increased Th17 differentiation. This metabolic reprogramming also enhances resistance to reactive oxygen species (ROS) and increases inosine accumulation. When challenged with tumors, GLUT1-overexpressing CAR-T cells secrete more proinflammatory cytokines, show enhanced cytotoxicity, and demonstrate superior tumor control and persistence in mouse models. The findings suggest that glucose availability is a limiting factor for CAR-T cell function, and enhancing glucose availability through GLUT1 overexpression could improve antitumor immune function. The study also shows that GLUT1 overexpression enhances metabolic pathways that favor resistance to ROS suppression, alters arginine and inosine metabolism, and improves CAR-T cell tumor clearance in vivo. Overall, the results demonstrate that GLUT1 overexpression enhances CAR-T cell potency and persistence, providing a new approach to improve the effectiveness of engineered T cell populations for antitumor targeting.