对基因毒性疗法的耐药性和肿瘤复发是胶质母细胞瘤（GBM）（一种侵袭性脑肿瘤）的标志。在这项研究中，我们通过综合基因组分析、患者来源的 GBM 模型中的全基因组遗传扰动筛选和独立验证线，研究了治疗后复发性 GBM 的功能驱动因素。与原发性 GBM 前身相比，在复发性肿瘤模型中发现了一致的特定遗传依赖性，并伴有突变负荷增加以及转录物和蛋白质表达差异。我们的观察结果表明，多层次的遗传反应可驱动肿瘤复发，并表明 PTP4A2（蛋白酪氨酸磷酸酶 4A2）是复发性 GBM 中自我更新、增殖和致瘤性的调节剂。 PTP4A2 的遗传扰动或小分子抑制通过具有迂回引导受体 1 (ROBO1) 及其下游分子参与者的去磷酸化轴发挥作用，利用对 ROBO 信号传导的功能依赖性。由于泛 PTP4A 抑制剂在体内穿过血脑屏障的渗透性较差，因此我们针对 ROBO1（一种在复发性 GBM 样本中富集的细胞表面受体）设计了第二代嵌合抗原受体 (CAR) T 细胞疗法。在复发性 GBM 细胞系来源的异种移植 (CDX) 模型中，单剂量 ROBO1 靶向 CAR T 细胞使中位存活率翻倍。此外，在成人肺脑转移和儿童复发性髓母细胞瘤的 CDX 模型中，ROBO1 CAR T 细胞消除了 50-100% 小鼠的肿瘤。我们的研究发现了一种有前景的多靶点 PTP4A-ROBO1 信号轴，可驱动复发性 GBM 的致瘤性，并在其他恶性脑肿瘤中具有潜力。© 2024。作者获得 Springer Nature America, Inc. 的独家许可。

Resistance to genotoxic therapies and tumor recurrence are hallmarks of glioblastoma (GBM), an aggressive brain tumor. In this study, we investigated functional drivers of post-treatment recurrent GBM through integrative genomic analyses, genome-wide genetic perturbation screens in patient-derived GBM models and independent lines of validation. Specific genetic dependencies were found consistent across recurrent tumor models, accompanied by increased mutational burden and differential transcript and protein expression compared to its primary GBM predecessor. Our observations suggest a multi-layered genetic response to drive tumor recurrence and implicate PTP4A2 (protein tyrosine phosphatase 4A2) as a modulator of self-renewal, proliferation and tumorigenicity in recurrent GBM. Genetic perturbation or small-molecule inhibition of PTP4A2 acts through a dephosphorylation axis with roundabout guidance receptor 1 (ROBO1) and its downstream molecular players, exploiting a functional dependency on ROBO signaling. Because a pan-PTP4A inhibitor was limited by poor penetrance across the blood-brain barrier in vivo, we engineered a second-generation chimeric antigen receptor (CAR) T cell therapy against ROBO1, a cell surface receptor enriched across recurrent GBM specimens. A single dose of ROBO1-targeted CAR T cells doubled median survival in cell-line-derived xenograft (CDX) models of recurrent GBM. Moreover, in CDX models of adult lung-to-brain metastases and pediatric relapsed medulloblastoma, ROBO1 CAR T cells eradicated tumors in 50-100% of mice. Our study identifies a promising multi-targetable PTP4A-ROBO1 signaling axis that drives tumorigenicity in recurrent GBM, with potential in other malignant brain tumors.© 2024. The Author(s), under exclusive licence to Springer Nature America, Inc.