Boardman Dominic A, Mangat Sonya, Gillies Jana K, Leon Lorna, Fung Vivian C W, Haque Manjurul, Mojibian Majid, Halvorson Torin, Huang Qing, Tuomela Karoliina, Wardell Christine M, Brown Andrew, Lam Avery J, Levings Megan K
Department of Surgery, The University of British Columbia, Vancouver, BC, Canada.
BC Children's Hospital Research Institute, Vancouver, BC, Canada.
Sci Adv. 2025 Jun 13;11(24):eadx7845. doi: 10.1126/sciadv.adx7845.
Regulatory T cell (T cell) therapy has been transformed through the use of chimeric antigen receptors (CARs). We previously found that human T cells minimally produce IL-10 and have a limited capacity to control innate immunity compared to type 1 regulatory T cells (T1 cells). To create "hybrid" CAR T cells with T1 cell-like properties, we examined whether the locus could be exploited to endow T cells with CAR-regulated IL-10 expression. CRISPR-mediated PD1 deletion increased CAR T cell activation, and knock-in of under control of the PD1 promoter resulted in CAR-induced IL-10 secretion. knock-in improved CAR T cell function, as determined by increased suppression of dendritic cells and alloantigen- and islet autoantigen-specific T cells. In vivo, knock-in CAR T cells were stable, safe, and suppressed dendritic cells and xenogeneic graft-versus-host disease. CRISPR-mediated engineering to simultaneously remove an inhibitory signal and enhance a suppressive mechanism is a previously unexplored approach to improve CAR T cell potency.
调节性T细胞（T细胞）疗法通过嵌合抗原受体（CAR）的使用而发生了变革。我们之前发现，与1型调节性T细胞（T1细胞）相比，人T细胞产生白细胞介素-10（IL-10）的量极少，并且控制先天免疫的能力有限。为了创建具有T1细胞样特性的“杂交”CAR T细胞，我们研究了是否可以利用该基因座赋予T细胞CAR调节的IL-10表达。CRISPR介导的程序性死亡蛋白1（PD1）缺失增加了CAR T细胞的活化，并且在PD1启动子控制下敲入该基因导致CAR诱导的IL-10分泌。该基因敲入改善了CAR T细胞功能，这通过对树突状细胞以及同种异体抗原和胰岛自身抗原特异性T细胞的抑制增加来确定。在体内，该基因敲入的CAR T细胞稳定、安全，并抑制树突状细胞和异种移植物抗宿主病。CRISPR介导的工程技术同时去除抑制信号并增强抑制机制是一种以前未探索过的提高CAR T细胞效力的方法。
