生物功能性水凝胶涂层膜减少 CAR-T 细胞生产中的 T 细胞耗竭

Hydrogel-coated membranes (HCMs) were designed with different co-stimulatory ligands and a physiologically-relevant substrate modulus inspired by the native microenvironment in which T cells are programmed. Phenotype, activation, and exhaustion markers were used to compare T cells cultured with HCMs or industry standard TransAct. Next, transduction with a CD19 CAR lentivirus was performed, and the killing potential of the resulting CAR T product was evaluated using an in vitro cytolysis model.

With this controlled and well-defined system, we hypothesized that a combination of ligands inspired by antigen-presenting cells would promote desired T-cell phenotypes with reduced exhaustion and thereby improved killing efficacy. We found memory phenotypes, minimal exhaustion, and similar activation profiles with HCMs. Additionally, increased T-cell transduction and decreased exhaustion for the CAR T population were observed with HCMs. Further, the killing potential of the resulting CAR T product was evaluated, finding improved in vitro cytolysis of target cells with lower variability with HCMs. DISCUSSION: These results demonstrate the importance of lower T-cell exhaustion in CAR T manufacturing and present significant opportunities to modulate T-cell phenotypes for cell therapy applications using engineered bioinspired materials that display combinations of co-stimulatory molecules.