高级别胶质瘤，包括胶质母细胞瘤（GBM）和弥漫性中线胶质瘤（DMG），是最致命和最具侵袭性的脑癌，目前的治疗方式疗效有限。嵌合抗原受体 (CAR) T 细胞疗法已成为一种有前景的策略，具有肿瘤特异性靶向和穿透血脑屏障的独特能力。然而，有效的临床应用取决于抗原的最佳选择，目前抗原的数量有限，目前正在研究中。我们对来自成人和儿童患者的原代人高级别胶质瘤样本进行了细胞表面蛋白质组学分析。这导致 Ephrin A 型受体 3 (EphA3) 被鉴定为普遍表达的靶标。我们设计了第二代 EphA3 靶向 CAR T 细胞，并使用 GBM 和 DMG 的体外和体内模型评估其功能。EphA3 靶向 CAR T 细胞在体外表现出对人 GBM 和 DMG 细胞系的强大抗原特异性杀伤作用。在原位异种移植 NSG 小鼠模型中，靶向 EphA3 的 CAR T 细胞不仅有效地根除肿瘤，而且还建立了对再攻击具有保护作用的功能性 T 细胞群。值得注意的是，接受第二次对侧原位肿瘤植入的小鼠重新获得了肿瘤的完全清除，并在初次治疗后 6 个月内保持了持续的完全缓解。基于 EphA3 抗体在临床环境中已被证实的安全性，我们的研究提供了令人信服的临床前证据，支持以下药物的功效：针对高级别神经胶质瘤的 EphA3 靶向 CAR T 细胞。这些发现强调了将这种创新疗法转化为临床试验的潜力，旨在彻底改变患有这些可怕脑癌的患者的治疗格局。© 作者（或其雇主）2024。CC 允许重复使用经过。英国医学杂志出版。

High-grade gliomas including glioblastoma (GBM) and diffuse midline gliomas (DMG) represent the most lethal and aggressive brain cancers where current treatment modalities offer limited efficacy. Chimeric antigen receptor (CAR) T cell therapies have emerged as a promising strategy, boasting tumor-specific targeting and the unique ability to penetrate the blood-brain barrier. However, the effective clinical application hinges on the optimal choice of antigen, with a limited number, currently under investigation.We employed cell surface proteomic analysis of primary human high-grade glioma samples from both adult and pediatric patients. This led to the identification of Ephrin type-A receptor 3 (EphA3) as a prevalently expressed target. We engineered a second-generation EphA3-targeted CAR T cell and assessed function using in vitro and in vivo models of GBM and DMG.EphA3-targeted CAR T cells demonstrated robust antigen-specific killing of human GBM and DMG cell lines in vitro. In an orthotopic xenograft NSG mouse model, EphA3-targeted CAR T cells not only effectively eradicated tumors but also established a functional T cell population protective on rechallenge. Remarkably, mice rechallenged with a second contralateral orthotopic tumor implantation achieved complete tumor clearance and maintained a sustained complete response 6 months following initial treatment.Building on the proven safety profile of EphA3 antibodies in clinical settings, our study provides compelling preclinical evidence supporting the efficacy of EphA3-targeted CAR T cells against high-grade gliomas. These findings underscore the potential for transitioning this innovative therapy into clinical trials, aiming to revolutionize the treatment landscape for patients afflicted with these formidable brain cancers.© Author(s) (or their employer(s)) 2024. Re-use permitted under CC BY. Published by BMJ.