Development of Ingenui-T, a Novel Vein-to-Vein Solution for Rapid Autologous CAR T-Cell Manufacturing Starting From Whole Blood, for the Treatment of Autoimmune Diseases

ABSTRACT Apheresis, a conventional starting point for manufacturing chimeric antigen receptor (CAR) T-cell therapy, poses challenges due to the length and invasiveness of the procedure, the high demand for and limited quantity of apheresis beds, and additional resource constraints at collection centers. Furthermore, traditional CAR T-cell manufacturing often involves extended cell culture periods, leading to a final product that has progressed through the differentiation process and contains a higher frequency of cells with phenotypes that are indicative of lower functionality or potency. 1 Here, we show that anti-CD19 CAR T-cells manufactured from fresh whole blood with minimal ex vivo expansion using Ingenui-T exhibit comparable or superior CAR-mediated and CD19-dependent functional activity compared to CAR T-cells manufactured from cryopreserved leukapheresis material following the a conventional manufacturing process. Anti-CD19 CAR T-cell production, manufactured from whole blood in less than 3 days of in vitro culture using Ingenui-T, yielded an average of about 40 million cells per 100 mL with high CD3 purity and viability. Furthermore, the final product was composed of cells with less differentiated phenotypes and sustained cytotoxic activity against CD19 + target cells at a lower dose than conventionally manufactured CAR T cells in preclinical in vitro assays. Ingenui-T is an innovative vein-to-vein solution, aimed at enhancing the patient experience, feasibility, and accessibility of CAR T-cell therapy by alleviating challenges linked to apheresis-based methods, with its patient-friendly nature, cost-effectiveness, and distinctive methodology.