Rejuvenating Effector/Exhausted CAR T Cells to Stem Cell Memory-Like CAR T Cells By Resting Them in the Presence of CXCL12 and the NOTCH Ligand.
Rejuvenating Effector/Exhausted CAR T Cells to Stem Cell Memory-Like CAR T Cells By Resting Them in the Presence of CXCL12 and the NOTCH Ligand.
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DOI：
10.1158/2767-9764.crc-21-0034
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发表时间：
2021-10
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具有干细胞记忆（TSCM）表型的T细胞为T细胞转移疗法提供长期和有效的抗肿瘤作用。尽管已经报道了用于体外诱导TSCM样细胞的各种方法，但是很少有方法从效应/耗尽的T细胞产生TSCM样细胞。我们已经报道了与表达Notch配体的OP 9基质细胞共培养诱导TSCM样（iTSCM）细胞。在这里，我们建立了一个无饲养层的培养系统，以改善从扩增的表达嵌合抗原受体（CAR）的T细胞产生iTSCM细胞;在IL 7、CXCL 12、IGF-I和Notch配体hDLL 1的存在下培养CAR T细胞。无饲养层的CAR-iTSCM细胞显示出与OP 9-hDLL 1饲养层细胞诱导的CAR-iTSCM细胞相似的细胞表面标志物和基因表达，包括SCM相关基因TCF 7、LEF 1和BCL 6的表达升高，以及耗竭相关基因如LAG 3、TOX和NR 4A 1的表达降低。与饲养细胞诱导的CAR-iTSCM细胞相比，无饲养细胞的CAR-iTSCM细胞显示出依赖于氧化磷酸化的更高增殖能力，并显示出更高的IL 2产生和更强的体内抗肿瘤活性。我们的无饲养层培养系统代表了一种将效应子/耗尽的CAR T细胞再生为SCM样CAR T细胞的方法。用我们定义的因子将静息CAR T细胞重新编程为SCM样状态，并能够开发改进的CAR T细胞疗法。
T cells with a stem cell memory (TSCM) phenotype provide long-term and potent antitumor effects for T-cell transfer therapies. Although various methods for the induction of TSCM-like cells in vitro have been reported, few methods generate TSCM-like cells from effector/exhausted T cells. We have reported that coculture with the Notch ligand–expressing OP9 stromal cells induces TSCM-like (iTSCM) cells. Here, we established a feeder-free culture system to improve iTSCM cell generation from expanded chimeric antigen receptor (CAR)-expressing T cells; culturing CAR T cells in the presence of IL7, CXCL12, IGF-I, and the Notch ligand, hDLL1. Feeder-free CAR-iTSCM cells showed the expression of cell surface markers and genes similar to that of OP9-hDLL1 feeder cell–induced CAR-iTSCM cells, including the elevated expression of SCM-associated genes, TCF7, LEF1, and BCL6, and reduced expression of exhaustion-associated genes like LAG3, TOX, and NR4A1. Feeder-free CAR-iTSCM cells showed higher proliferative capacity depending on oxidative phosphorylation and exhibited higher IL2 production and stronger antitumor activity in vivo than feeder cell–induced CAR-iTSCM cells. Our feeder-free culture system represents a way to rejuvenate effector/exhausted CAR T cells to SCM-like CAR T cells. Resting CAR T cells with our defined factors reprograms exhausted state to SCM-like state and enables development of improved CAR T-cell therapy.