Chimeric antigen receptor (CAR) T cells targeting receptors on tumour cells have had limited success in patients with glioblastoma. Here we report the development and therapeutic performance of CAR constructs leveraging protein binders computationally designed de novo to have high affinity for the epidermal growth factor receptor (EGFR) or the tumour-associated antigen CD276, which are overexpressed in glioblastoma. With respect to T cells with a CAR using an antibody-derived single-chain variable fragment as antigen-binding domain, the designed binders on CAR T cells promoted the proliferation of the cells, the secretion of cytotoxic cytokines and their resistance to cell exhaustion, and improved antitumour performance in vitro and in vivo. Moreover, CARs with the binders exhibited higher surface expression and greater resistance to degradation, as indicated by bulk and single-cell transcriptional profiling of the cells. The de novo design of binding domains for specific tumour antigens may potentiate the antitumour efficacy of CAR T cell therapies for other solid cancers.

靶向肿瘤细胞受体的嵌合抗原受体 （CAR） T 细胞在胶质母细胞瘤患者中的成功有限。在这里，我们报告了 CAR 构建体的开发和治疗性能，这些构建体利用计算设计的蛋白质结合剂从头设计，对表皮生长因子受体 （EGFR） 或肿瘤相关抗原 CD276 具有高亲和力，它们在胶质母细胞瘤中过表达。对于使用抗体衍生的单链可变片段作为抗原结合结构域的 CAR 的 T 细胞，CAR T 细胞上设计的结合剂促进了细胞增殖、细胞毒性细胞因子的分泌及其对细胞耗竭的抵抗力，并提高了体外和体内的抗肿瘤性能。此外，带有结合剂的 CAR 表现出更高的表面表达和更强的抗降解性，如细胞的大量和单细胞转录谱所示。特异性肿瘤抗原结合结构域的从头设计可能会增强 CAR T 细胞疗法对其他实体癌的抗肿瘤疗效。