Patient‑specific profiling of TCR‑T cell efficacy enabled by a rapid and standardized reporter system

T cell receptor-engineered T (TCR-T) cell therapy has shown promising clinical potential, yet accurate assessment of patient suitability remains challenging because current genomic and transcriptomic approaches do not directly capture physiological antigen presentation or authentic T-cell responsiveness. Here, we developed a rapid and standardized Jurkat-based reporter platform, termed the TCR-SignalFunc system, for functional evaluation of TCR-T cell therapeutic potential. This system was established by eliminating endogenous TCR expression, incorporating a highly sensitive synthetic 8×NFAT:AP-1 promoter for activation sensing, and introducing CD8 to better recapitulate the functional context of therapeutic TCR-T cells. TCR activation was quantified using the SignalFunc Index, defined as the percentage of ZsGreen-positive cells. The platform demonstrated robust sensitivity, rapid signal detection, and strong correlation with the cytotoxic activity of corresponding TCR-T cells across multiple antigen specificities in vitro and in vivo. Importantly, we further adapted this system for direct application to patient-derived ex vivo tissues by integrating tissue dissociation, tumor-cell proportion estimation, and standardized co-culture assays, enabling patient-specific functional stratification within 12 hours. In colorectal cancer samples, the system accurately distinguished tumor-specific responsiveness of TCRs targeting p53^R175H/HLA-A0201 and CEA/HLA-A0201. Collectively, this work establishes a practical and scalable functional platform for rapid assessment of TCR-T cell efficacy, with broad potential for patient selection, therapeutic evaluation, and future TCR optimization.

A rapid, standardized reporter system enables patient-specific profiling of TCR-T cell efficacy and facilitates assessment of therapeutic potential in ex vivo tissues for personalized immunotherapy evaluation.