Enhancing Temozolomide (TMZ) Chemosensitivity Using CRISPR-dCas9-mediated Downregulation of O-methylguanine DNA Methyltransferase (MGMT)

Overview

Journal J Neurooncol

Publisher Springer

Date 2024 May 19

PMID 38762829

Authors

Yasamin Yousefi

Reza Nejati

Atiye Eslahi

Farzaneh Alizadeh

Shima Farrokhi

Ahmad Asoodeh

Majid Mojarrad

Affiliations

Soon will be listed here.

Abstract

Purpose: Glioblastoma (GBM) stands out as the most prevalent and aggressive intracranial tumor, notorious for its poor prognosis. The current standard-of-care for GBM patients involves surgical resection followed by radiotherapy, combined with concurrent and adjuvant chemotherapy using Temozolomide (TMZ). The effectiveness of TMZ primarily relies on the activity of O-methylguanine DNA methyltransferase (MGMT), which removes alkyl adducts from the O position of guanine at the DNA level, thereby counteracting the toxic effects of TMZ.

Method: In this study, we employed fusions of catalytically-inactive Cas9 (dCas9) to DNA methyltransferases (dCas9-DNMT3A) to selectively downregulation MGMT transcription by inducing methylation at MGMT promoter and K-M enhancer.

Result: Our findings demonstrate a significant reduction in MGMT expression, leading to intensified TMZ sensitivity in the HEK293T cell line.

Conclusion: This study serves as a proof of concept for the utilization of CRISPR-based gene suppression to overcome TMZ resistance and enhance the lethal effect of TMZ in glioblastoma tumor cells.

Citing Articles

PDCD10 Is a Key Player in TMZ-Resistance and Tumor Cell Regrowth: Insights into Its Underlying Mechanism in Glioblastoma Cells.

Zhu Y, Kim S, Chen Z, Will R, Zhong R, Dammann P Cells. 2024; 13(17.

PMID: 39273014 PMC: 11394141. DOI: 10.3390/cells13171442.

References

Soeda A, Hara A, Kunisada T, Yoshimura S, Iwama T, Park D . The evidence of glioblastoma heterogeneity. Sci Rep. 2015; 5:7979. PMC: 4306917. DOI: 10.1038/srep07979. View

Becker A, Sells B, Haque S, Chakravarti A . Tumor Heterogeneity in Glioblastomas: From Light Microscopy to Molecular Pathology. Cancers (Basel). 2021; 13(4). PMC: 7918815. DOI: 10.3390/cancers13040761. View

Rabah N, Ait Mohand F, Kravchenko-Balasha N . Understanding Glioblastoma Signaling, Heterogeneity, Invasiveness, and Drug Delivery Barriers. Int J Mol Sci. 2023; 24(18). PMC: 10532387. DOI: 10.3390/ijms241814256. View

Meyer M . Malignant gliomas in adults. N Engl J Med. 2008; 359(17):1850. DOI: 10.1056/NEJMc086380. View

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

Kanzawa T, Bedwell J, Kondo Y, Kondo S, Germano I . Inhibition of DNA repair for sensitizing resistant glioma cells to temozolomide. J Neurosurg. 2004; 99(6):1047-52. DOI: 10.3171/jns.2003.99.6.1047. View

Nakada M, Furuta T, Hayashi Y, Minamoto T, Hamada J . The strategy for enhancing temozolomide against malignant glioma. Front Oncol. 2012; 2:98. PMC: 3418701. DOI: 10.3389/fonc.2012.00098. View

Nagel Z, Kitange G, Gupta S, Joughin B, Chaim I, Mazzucato P . DNA Repair Capacity in Multiple Pathways Predicts Chemoresistance in Glioblastoma Multiforme. Cancer Res. 2016; 77(1):198-206. PMC: 6100738. DOI: 10.1158/0008-5472.CAN-16-1151. View

Hombach-Klonisch S, Mehrpour M, Shojaei S, Harlos C, Pitz M, Hamai A . Glioblastoma and chemoresistance to alkylating agents: Involvement of apoptosis, autophagy, and unfolded protein response. Pharmacol Ther. 2017; 184:13-41. DOI: 10.1016/j.pharmthera.2017.10.017. View

10.

Brandes A, Franceschi E, Paccapelo A, Tallini G, de Biase D, Ghimenton C . Role of Methylation Status at Time of Diagnosis and Recurrence for Patients with Glioblastoma: Clinical Implications. Oncologist. 2017; 22(4):432-437. PMC: 5388380. DOI: 10.1634/theoncologist.2016-0254. View

11.

Moradi Binabaj M, Bahrami A, Shahidsales S, Joodi M, Joudi Mashhad M, Hassanian S . The prognostic value of MGMT promoter methylation in glioblastoma: A meta-analysis of clinical trials. J Cell Physiol. 2017; 233(1):378-386. DOI: 10.1002/jcp.25896. View

12.

Dolan M, Mitchell R, Mummert C, Moschel R, Pegg A . Effect of O6-benzylguanine analogues on sensitivity of human tumor cells to the cytotoxic effects of alkylating agents. Cancer Res. 1991; 51(13):3367-72. View

13.

Quinn J, Jiang S, Reardon D, Desjardins A, Vredenburgh J, Rich J . Phase II trial of temozolomide plus o6-benzylguanine in adults with recurrent, temozolomide-resistant malignant glioma. J Clin Oncol. 2009; 27(8):1262-7. PMC: 2667825. DOI: 10.1200/JCO.2008.18.8417. View

14.

Aasland D, Reich T, Tomicic M, Switzeny O, Kaina B, Christmann M . Repair gene O -methylguanine-DNA methyltransferase is controlled by SP1 and up-regulated by glucocorticoids, but not by temozolomide and radiation. J Neurochem. 2017; 144(2):139-151. DOI: 10.1111/jnc.14262. View

15.

Happold C, Stojcheva N, Silginer M, Weiss T, Roth P, Reifenberger G . Transcriptional control of O -methylguanine DNA methyltransferase expression and temozolomide resistance in glioblastoma. J Neurochem. 2018; 144(6):780-790. DOI: 10.1111/jnc.14326. View

16.

Wang K, Chen D, Qian Z, Cui D, Gao L, Lou M . Hedgehog/Gli1 signaling pathway regulates MGMT expression and chemoresistance to temozolomide in human glioblastoma. Cancer Cell Int. 2017; 17:117. PMC: 5715541. DOI: 10.1186/s12935-017-0491-x. View

17.

Hegi M, Diserens A, Gorlia T, Hamou M, De Tribolet N, Weller M . MGMT gene silencing and benefit from temozolomide in glioblastoma. N Engl J Med. 2005; 352(10):997-1003. DOI: 10.1056/NEJMoa043331. View

19.

Kitange G, Mladek A, Carlson B, Schroeder M, Pokorny J, Cen L . Inhibition of histone deacetylation potentiates the evolution of acquired temozolomide resistance linked to MGMT upregulation in glioblastoma xenografts. Clin Cancer Res. 2012; 18(15):4070-9. PMC: 3716364. DOI: 10.1158/1078-0432.CCR-12-0560. View

20.

Chen X, Zhang M, Gan H, Wang H, Lee J, Fang D . A novel enhancer regulates MGMT expression and promotes temozolomide resistance in glioblastoma. Nat Commun. 2018; 9(1):2949. PMC: 6063898. DOI: 10.1038/s41467-018-05373-4. View

21.

Harris L, Remack J, Brent T . Identification of a 59 bp enhancer located at the first exon/intron boundary of the human O6-methylguanine DNA methyltransferase gene. Nucleic Acids Res. 1994; 22(22):4614-9. PMC: 308508. DOI: 10.1093/nar/22.22.4614. View