Kavli Affiliate: Tejal Desai
| Authors: Qinghe Zeng, Landon Flemming, Yuanzhou Chen, Thomas Mazumder, Heinz Hammerlindl, Greg M. Allen, Ricardo Almeida, Jasper Z. Williams, Rogelio A. Hernandez-Lopez, Justin Eyquem, Chun Jimmie Ye, Wendell A Lim, Qizhi Tang, Tejal Desai and Xiao Huang
| Summary:
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.