Bradshaw, Gary Andrew

Abstract Determining the balance between DNA double strand break repair (DSBR) pathways is essential for understanding treatment response in cancer. We report a method for simultaneously measuring non-homologous end joining (NHEJ), homologous recombination (HR), and microhomology-mediated end joining (MMEJ) and show that HR and MMEJ are therapeutically-targetable treatment resistance mechanisms in glioblastoma. Using a targeted screen of clinically relevant small molecules, we identify the ATM kinase inhibitor, AZD1390, as a potent dual HR/MMEJ inhibitor. Mechanistic studies in GBM cell lines show that AZD1390 suppresses DSB end resection and blocks radiation-induced phosphorylation of DSBR proteins including nibrin, Rad50, and Mettl16. Interestingly, AZD1390 enhances the cytotoxic effects of TMZ and radiation in both treatment-naïve and treatment-resistant GBM samples harboring mutations in the TP53 gene. We show that TP53 mutation is associated with upregulation of ATM pathway genes in GBM patient samples along with heightened ATM-dependent HR and MMEJ in GBM cell lines. Finally, we show that TP53-mutant GBMs fail to arrest in G2 phase after radiation/AZD1390 or TMZ/AZD1390 combinations and instead attempt mitosis, leading to mitotic cell death. Expression of the mitotic DSBR factor, RHINO1, is associated with poor prognosis in patients with TP53-mutant GBMs, suggesting that factors that promote efficient mitotic DSBR protect TP53-mutant GBMs from treatment-induced cell death. This report identifies ATM-dependent HR and MMEJ as targetable resistance mechanisms in TP53-mutant GBM and establishes ATM inhibition as a potential strategy for overcoming intrinsic and acquired treatment resistance in TP53-mutant GBMs. Citation Format: Daniel J Laverty, Shiv Gupta, Gary Bradshaw, Jiajia Chen, Brett L Carlson, Nery Matias Calmo, Shulan Tian, Jann N Sarkaria, Zachary D Nagel. ATM inhibitors exploit checkpoint defects and ATM-dependent double strand break repair to induce mitotic cell death in TP53-mutant glioblastoma [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Functional and Genomic Precision Medicine in Cancer: Different Perspectives, Common Goals; 2025 Mar 11-13; Boston, MA. Philadelphia (PA): AACR; Cancer Res 2025;85(5 Suppl):Abstract nr A011.