Although chimeric antigen receptor (CAR) T cells have demonstrated signs of antitumor activity against glioblastoma (GBM), tumor heterogeneity remains a critical challenge. To achieve broader and more effective GBM targeting, we developed a peptide-bearing CAR exploiting the GBM-binding potential of chlorotoxin (CLTX). We find that CLTX peptide binds a great proportion of tumors and constituent tumor cells. CAR T cells using CLTX as the targeting domain (CLTX-CAR T cells) mediate potent anti-GBM activity and efficiently target tumors lacking expression of other GBM-associated antigens. Treatment with CLTX-CAR T cells resulted in tumor regression in orthotopic xenograft GBM tumor models. CLTX-CAR T cells do not exhibit observable off-target effector activity against normal cells or after adoptive transfer into mice. Effective targeting by CLTX-CAR T cells requires cell surface expression of matrix metalloproteinase–2. Our results pioneer a peptide toxin in CAR design, expanding the repertoire of tumor-selective CAR T cells with the potential to reduce antigen escape.

尽管嵌合抗原受体 (CAR) T 细胞已显示出抗胶质母细胞瘤 (GBM) 的抗肿瘤活性迹象，但肿瘤异质性仍然是一个关键挑战。为了实现更广泛和更有效的 GBM 靶向，我们开发了一种带有肽的 CAR，利用氯毒素 (CLTX) 的 GBM 结合潜力。我们发现 CLTX 肽结合了很大比例的肿瘤和组成肿瘤细胞。使用 CLTX 作为靶向域的 CAR T 细胞（CLTX-CAR T 细胞）介导有效的抗 GBM 活性并有效靶向缺乏其他 GBM 相关抗原表达的肿瘤。用 CLTX-CAR T 细胞治疗导致原位异种移植 GBM 肿瘤模型中的肿瘤消退。CLTX-CAR T 细胞对正常细胞或过继转移到小鼠体内后没有表现出可观察到的脱靶效应活性。CLTX-CAR T 细胞的有效靶向需要基质金属蛋白酶-2 的细胞表面表达。我们的研究结果开创了 CAR 设计中的肽毒素，扩大了肿瘤选择性 CAR T 细胞的库，并有可能减少抗原逃逸。