胶质母细胞瘤具有高度浸润性生长模式，导致复发和生存率低。尽管浸润是一个关键的治疗目标，但临床上不存在对抗胶质母细胞瘤侵袭的有用疗法。在这里，我们报告抑制共济失调毛细血管扩张和 Rad 3 相关激酶 (ATR) 通过细胞骨架网络失调和随后的整合素运输来减少胶质母细胞瘤细胞的侵袭。在体外和体内评估了 ATR 抑制反应的胶质母细胞瘤运动和侵袭能力 (ATRi) ）和使用延时显微镜、两种原位胶质母细胞瘤模型和活体成像的 ATR 过度表达。通过高通量、超分辨率成像和活体成像研究了细胞骨架网络和内吞处理的破坏。在临床数据集中，高 ATR 表达与显着较差的生存率相关，而使用胶质母细胞瘤肿瘤标本的组织学、蛋白质表达和空间转录组学显示，在浸润性组织中 ATR 表达较高边距。两种不同化合物的药理抑制和针对 ATR 的 RNAi 可以抵抗胶质母细胞瘤的侵袭，而 ATR 的过度表达则促进迁移。随后的研究表明，细胞骨架失调减少了生长锥样结构中整合素的巨胞饮内化，导致肿瘤微管回缩缺陷。使用胶质母细胞瘤的两种原位体内模型和活体成像证实了这些发现的生物学相关性和转化潜力。我们证明了 ATR 在确定胶质母细胞瘤细胞侵袭方面的新作用，并提出 ATR 的药理学靶向可能具有深远的临床益处，超越放射增敏。© 作者 2023。由牛津大学出版社代表神经肿瘤学会出版。

Glioblastomas have highly infiltrative growth patterns that contribute to recurrence and poor survival. Despite infiltration being a critical therapeutic target, no clinically useful therapies exist which counter glioblastoma invasion. Here, we report that inhibition of Ataxia telangiectasia and Rad 3 related kinase (ATR) reduces invasion of glioblastoma cells through dysregulation of cytoskeletal networks and subsequent integrin trafficking.Glioblastoma motility and invasion were assessed in vitro and in vivo in response to ATR inhibition (ATRi) and ATR over-expression using timelapse microscopy, two orthotopic glioblastoma models and intravital imaging. Disruption to cytoskeleton networks and endocytic processing were investigated via high throughput, super resolution imaging and intravital imaging.High ATR expression was associated with significantly poorer survival in clinical datasets whilst histological, protein expression and spatial transcriptomics using glioblastoma tumour specimens revealed higher ATR expression at infiltrative margins. Pharmacological inhibition with two different compounds and RNAi targeting of ATR opposed invasion of glioblastoma, whereas overexpression of ATR drove migration. Subsequent investigation revealed that cytoskeletal dysregulation reduced macropinocytotic internalisation of integrins at growth cone-like structures, resulting in a tumour microtube retraction defect. The biological relevance and translational potential of these findings was confirmed using two orthotopic in vivo models of glioblastoma and intravital imaging.We demonstrate a novel role for ATR in determining invasion in glioblastoma cells and propose that pharmacological targeting of ATR could have far reaching clinical benefits beyond radiosensitisation.© The Author(s) 2023. Published by Oxford University Press on behalf of the Society for Neuro-Oncology.