Programmable microparticles rewire CAR signaling to enable super-physiological expansion of human T cells in vitro
Zeng, Q.; Flemming, L.; Chen, Y.; Mazumder, T.; Hammerlindl, H.; Allen, G. M.; Almeida, R.; Williams, J. Z.; Hernandez-Lopez, R. A.; Eyquem, J.; Ye, C. J.; Lim, W. A.; Tang, Q.; Desai, T.; Huang, X.
Show abstract
T cell proliferative capacity and persistence critically determine the therapeutic success of chimeric antigen receptor (CAR) T cells. However, it remains unknown if and how human CAR-T cells can be externally programmed to reach maximal proliferative capacity. Here, we use programmable PLGA microparticles functionalized with CAR-antigens and CD28-costimulatory antibodies (CAREp) to repeatedly stimulate human CD8+ CAR-T cells in vitro. CAREp-stimulated CAR-T cells expanded continuously for over 100 days--versus [~]30 days with tumor cell stimulation--and achieved up to 1018-fold cumulative expansion, greatly surpassing CD3/28-Dynabeads. Early-phase transcriptomic responses-- upregulation of DNA repair, cell cycle, telomere maintenance, and mitochondrial pathways--aligned with long-term outcomes: massive proliferation, telomere stability, robust respiration, and preserved progenitor phenotype by single-cell sequencing. Differentiation and exhaustion signals were broadly suppressed. Transient telomerase activity further supported physiologic expansion. These findings demonstrate that nanoscale-controlled extracellular cues can rewire intracellular signaling to drive durable, super-physiological expansion of functional CAR-T cells.
Matching journals
The top 6 journals account for 50% of the predicted probability mass.