MGMT Function Determines the Differential Response of ATR Inhibitors with DNA-damaging Agents in Glioma Stem Cells for GBM Therapy

Overview

Journal Neurooncol Adv

Publisher Oxford University Press

Date 2024 Jan 12

PMID 38213834

Authors

Vincent W S Leong

Sabbir Khan

Pratibha Sharma

Shaofang Wu

Riya R Thomas

Xiaolong Li

Sanjay K Singh

Frederick F Lang

Alfred W K Yung

Dimpy Koul

Affiliations

Soon will be listed here.

Abstract

Background: The most prevalent cancer treatments cause cell death through DNA damage. However, DNA damage response (DDR) repair pathways, initiated by tumor cells, can withstand the effects of anticancer drugs, providing justification for combining DDR inhibitors with DNA-damaging anticancer treatments.

Methods: Cell viability assays were performed with CellTiter-Glo assay. DNA damage was evaluated using Western blotting analysis. RNA-seq and single-cell level expression were used to identify the DDR signatures. In vivo, studies were conducted in mice to determine the effect of ATris on TMZ sensitization.

Results: We found a subpopulation of glioma sphere-forming cells (GSCs) with substantial synergism with temozolomide (TMZ) using a panel of 3 clinical-grade ataxia-telangiectasia- and Rad3-related kinase inhibitors (ATRis), (elimusertib, berzosertib, and ceralasertib). Interestingly, most synergistic cell lines had O6-methylguanine-DNA methyltransferase (MGMT) promoter methylation, indicating that ATRi mainly benefits tumors with no MGMT repair. Further, TMZ activated the ATR-checkpoint kinase 1 (Chk1) axis in an MGMT-dependent way. TMZ caused ATR-dependent Chk1 phosphorylation and DNA double-strand breaks as shown by increased γH2AX. Increased DNA damage and decreased Chk1 phosphorylation were observed upon the addition of ATRis to TMZ in MGMT-methylated (MGMT-) GSCs. TMZ also improved sensitivity to ATRis in vivo, as shown by increased mouse survival with the TMZ and ATRi combination treatment.

Conclusions: This research provides a rationale for selectively targeting MGMT-methylated cells using ATRis and TMZ combination. Overall, we believe that MGMT methylation status in GBM could serve as a robust biomarker for patient selection for ATRi combined with TMZ.

References

Esteller M, Hamilton S, Burger P, Baylin S, Herman J . Inactivation of the DNA repair gene O6-methylguanine-DNA methyltransferase by promoter hypermethylation is a common event in primary human neoplasia. Cancer Res. 1999; 59(4):793-7. View

Vassin V, Anantha R, Sokolova E, Kanner S, Borowiec J . Human RPA phosphorylation by ATR stimulates DNA synthesis and prevents ssDNA accumulation during DNA-replication stress. J Cell Sci. 2009; 122(Pt 22):4070-80. PMC: 2776501. DOI: 10.1242/jcs.053702. View

Wu S, Wang S, Zheng S, Verhaak R, Koul D, Yung W . MSK1-Mediated β-Catenin Phosphorylation Confers Resistance to PI3K/mTOR Inhibitors in Glioblastoma. Mol Cancer Ther. 2016; 15(7):1656-68. PMC: 4936927. DOI: 10.1158/1535-7163.MCT-15-0857. View

Pearl L, Schierz A, Ward S, Al-Lazikani B, Pearl F . Therapeutic opportunities within the DNA damage response. Nat Rev Cancer. 2015; 15(3):166-80. DOI: 10.1038/nrc3891. View

Aasland D, Gotzinger L, Hauck L, Berte N, Meyer J, Effenberger M . Temozolomide Induces Senescence and Repression of DNA Repair Pathways in Glioblastoma Cells via Activation of ATR-CHK1, p21, and NF-κB. Cancer Res. 2018; 79(1):99-113. DOI: 10.1158/0008-5472.CAN-18-1733. View

Natsume A, Wakabayashi T, Ishii D, Maruta H, Fujii M, Shimato S . A combination of IFN-beta and temozolomide in human glioma xenograft models: implication of p53-mediated MGMT downregulation. Cancer Chemother Pharmacol. 2007; 61(4):653-9. DOI: 10.1007/s00280-007-0520-x. View

Clarke J, Iwamoto F, Sul J, Panageas K, Lassman A, DeAngelis L . Randomized phase II trial of chemoradiotherapy followed by either dose-dense or metronomic temozolomide for newly diagnosed glioblastoma. J Clin Oncol. 2009; 27(23):3861-7. PMC: 2727290. DOI: 10.1200/JCO.2008.20.7944. View

Lal S, Lacroix M, Tofilon P, Fuller G, Sawaya R, Lang F . An implantable guide-screw system for brain tumor studies in small animals. J Neurosurg. 2000; 92(2):326-33. DOI: 10.3171/jns.2000.92.2.0326. View

Ujifuku K, Mitsutake N, Takakura S, Matsuse M, Saenko V, Suzuki K . miR-195, miR-455-3p and miR-10a( *) are implicated in acquired temozolomide resistance in glioblastoma multiforme cells. Cancer Lett. 2010; 296(2):241-8. DOI: 10.1016/j.canlet.2010.04.013. View

10.

Lee S . Temozolomide resistance in glioblastoma multiforme. Genes Dis. 2018; 3(3):198-210. PMC: 6150109. DOI: 10.1016/j.gendis.2016.04.007. View

11.

Caldecott K . Protein ADP-ribosylation and the cellular response to DNA strand breaks. DNA Repair (Amst). 2014; 19:108-13. DOI: 10.1016/j.dnarep.2014.03.021. View

12.

Kwok M, Davies N, Agathanggelou A, Smith E, Petermann E, Yates E . Synthetic lethality in chronic lymphocytic leukaemia with DNA damage response defects by targeting the ATR pathway. Lancet. 2015; 385 Suppl 1:S58. DOI: 10.1016/S0140-6736(15)60373-7. View

13.

Stupp R, Mason W, van den Bent M, Weller M, Fisher B, Taphoorn M . Radiotherapy plus concomitant and adjuvant temozolomide for glioblastoma. N Engl J Med. 2005; 352(10):987-96. DOI: 10.1056/NEJMoa043330. View

14.

Ganesa S, Sule A, Sundaram R, Bindra R . Mismatch repair proteins play a role in ATR activation upon temozolomide treatment in MGMT-methylated glioblastoma. Sci Rep. 2022; 12(1):5827. PMC: 8987098. DOI: 10.1038/s41598-022-09614-x. View

15.

Quan R, Zhang H, Li Z, Li X . Survival analysis of patients with glioblastoma treated by long-term administration of temozolomide. Medicine (Baltimore). 2020; 99(2):e18591. PMC: 6959873. DOI: 10.1097/MD.0000000000018591. View

16.

Jackson C, Noorbakhsh S, Sundaram R, Kalathil A, Ganesa S, Jia L . Temozolomide Sensitizes MGMT-Deficient Tumor Cells to ATR Inhibitors. Cancer Res. 2019; 79(17):4331-4338. PMC: 6810597. DOI: 10.1158/0008-5472.CAN-18-3394. View

17.

Tian H, Gao Z, Li H, Zhang B, Wang G, Zhang Q . DNA damage response--a double-edged sword in cancer prevention and cancer therapy. Cancer Lett. 2014; 358(1):8-16. DOI: 10.1016/j.canlet.2014.12.038. View

18.

Zhang N, Wu X, Yang L, Xiao F, Zhang H, Zhou A . FoxM1 inhibition sensitizes resistant glioblastoma cells to temozolomide by downregulating the expression of DNA-repair gene Rad51. Clin Cancer Res. 2012; 18(21):5961-71. PMC: 3639123. DOI: 10.1158/1078-0432.CCR-12-0039. View

19.

van den Bent M, Dubbink H, Sanson M, van der Lee-Haarloo C, Hegi M, Jeuken J . MGMT promoter methylation is prognostic but not predictive for outcome to adjuvant PCV chemotherapy in anaplastic oligodendroglial tumors: a report from EORTC Brain Tumor Group Study 26951. J Clin Oncol. 2009; 27(35):5881-6. PMC: 2793037. DOI: 10.1200/JCO.2009.24.1034. View

20.

Esteller M, Garcia-Foncillas J, Andion E, Goodman S, Hidalgo O, Vanaclocha V . Inactivation of the DNA-repair gene MGMT and the clinical response of gliomas to alkylating agents. N Engl J Med. 2000; 343(19):1350-4. DOI: 10.1056/NEJM200011093431901. View