Framework humanization enhances GM3(Neu5Gc)-targeting CAR-T cell function by reducing tonic signaling.

ABSTRACT: GM3(Neu5Gc), a tumor-associated ganglioside absent in normal human tissues due to a CMP-N-acetylneuraminic acid hydroxylase (CMAH) mutation, is an attractive target for solid tumor immunotherapy. To advance the clinical potential of GM3(Neu5Gc)-targeted CAR-T cells, we systematically evaluated antibody humanization by comparing CARs based on the murine 14F7 antibody and its humanized variant 14F7hT. Within each scFv framework, we further assessed three hinge domains (CD8α, CD28, IgG4) to optimize CAR design. While hinge selection influenced in vitro cytotoxicity-favoring CD28-the humanized 14F7hT-based CARs consistently outperformed their murine counterparts in vivo. The optimized 14F7hT-CD28 CAR-T cells demonstrated superior expansion, persistence, tumor infiltration, and an

SUBMITTER: Tu J 

PROVIDER: S-EPMC12589110 | biostudies-literature | 2025

REPOSITORIES: biostudies-literature