嵌合抗原受体（CAR）T细胞疗法在治疗B细胞恶性肿瘤患者中取得的巨大成功促使其转化为实体瘤。就胶质母细胞瘤 (GBM) 而言，临床试验显示出有限的疗效，但开发更有效的抗 GBM CAR T 细胞的努力仍在继续。在本研究中，我们选择PTPRZ1作为GBM治疗的靶点。我们从人类噬菌体展示库中分离出 6 种抗人 PTPRZ1 scFv，并以 RNA 形式生产了第二代 CAR T 细胞。使用患者来源的 GBM PTPRZ1 敲入细胞系来选择具有高细胞毒性、同时始终显示高 CAR 表达的 CAR 构建体 (471_28z)。掺入 471_28z 的 CAR T 细胞能够释放 IFN-γ、IL-2、TNF-α、颗粒酶 B、IL-17A、IL-6 和可溶性 FasL，并表现出低强度信号传导。此外，它们在体外杀死后仍保持效应记忆表型。此外，471_28z CAR T细胞在被PTPRZ1阳性肿瘤细胞预激活后对PTPRZ1阴性细胞系表现出强烈的旁观者杀伤作用，但不会杀死抗原阴性非肿瘤细胞。在使用 NSG 小鼠的原位异种移植肿瘤模型中，单剂量的抗 PTPRZ1 CAR T 细胞显着延迟了肿瘤生长。总而言之，这些结果验证了 PTPRZ1 作为 GBM 靶点，并促进了抗 PTPRZ1 CAR T 细胞的临床转化。

The great success of chimeric antigen receptor (CAR) T-cell therapy in the treatment of patients with B-cell malignancies has prompted its translation to solid tumors. In the case of glioblastoma (GBM), clinical trials have shown modest efficacy, but efforts to develop more effective anti-GBM CAR T cells are ongoing. In this study, we selected PTPRZ1 as a target for GBM treatment. We isolated six anti-human PTPRZ1 scFv from a human phage display library and produced 2nd generation CAR T cells in an RNA format. Patient-derived GBM PTPRZ1-knock-in cell lines were used to select the CAR construct that showed high cytotoxicity while consistently displaying high CAR expression (471_28z). CAR T cells incorporating 471_28z were able to release IFN-γ, IL-2, TNF-α, Granzyme B, IL-17A, IL-6, and soluble FasL, and displayed low tonic signaling. Additionally, they maintained an effector memory phenotype after in vitro killing. In addition, 471_28z CAR T cells displayed strong bystander killing against PTPRZ1-negative cell lines after pre-activation by PTPRZ1-positive tumor cells but did not kill antigen-negative non-tumor cells. In an orthotopic xenograft tumor model using NSG mice, a single dose of anti-PTPRZ1 CAR T cells significantly delayed tumor growth. Taken together, these results validate PTPRZ1 as a GBM target and prompt the clinical translation of anti-PTPRZ1 CAR T cells.