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  3. A robust luminescent assay for screening alkyladenine DNA glycosylase inhibitors to overcome DNA repair and temozolomide drug resistance

A robust luminescent assay for screening alkyladenine DNA glycosylase inhibitors to overcome DNA repair and temozolomide drug resistance

  • J Pharm Anal. 2023 May;13(5):514-522. doi: 10.1016/j.jpha.2023.04.010.
Ying-Qi Song 1,  Guo-Dong Li 1,  Dou Niu 1,  Feng Chen 1,  Shaozhen Jing 2,  Vincent Kam Wai Wong 3,  Wanhe Wang 2,  Chung-Hang Leung 1 4
Affiliations

Affiliations

  • 1 State Key Laboratory of Quality Research in Chinese Medicine, Institute of Chinese Medical Sciences, University of Macau, Macao, 999078, China.
  • 2 Institute of Medical Research, Northwestern Polytechnical University, Xi'an, 710072, China.
  • 3 Dr. Neher's Biophysics Laboratory for Innovative Drug Discovery, State Key Laboratory of Quality Research in Chinese Medicine, Macau University of Science and Technology, Macao, 999078, China.
  • 4 Department of Biomedical Sciences, Faculty of Health Sciences, University of Macau, Macao, 999078, China.
PMID: 37305785 DOI: 10.1016/j.jpha.2023.04.010
Abstract

Temozolomide (TMZ) is an Anticancer agent used to treat glioblastoma, typically following radiation therapy and/or surgical resection. However, despite its effectiveness, at least 50% of patients do not respond to TMZ, which is associated with repair and/or tolerance of TMZ-induced DNA lesions. Studies have demonstrated that alkyladenine DNA glycosylase (AAG), an enzyme that triggers the base excision repair (BER) pathway by excising TMZ-induced N3-methyladenine (3meA) and N7-methylguanine lesions, is overexpressed in glioblastoma tissues compared to normal tissues. Therefore, it is essential to develop a rapid and efficient screening method for AAG inhibitors to overcome TMZ resistance in glioblastomas. Herein, we report a robust time-resolved photoluminescence platform for identifying AAG inhibitors with improved sensitivity compared to conventional steady-state spectroscopic methods. As a proof-of-concept, this assay was used to screen 1440 food and drug administration-approved drugs against AAG, resulting in the repurposing of sunitinib as a potential AAG inhibitor. Sunitinib restored glioblastoma (GBM) Cancer cell sensitivity to TMZ, inhibited GBM cell proliferation and stem cell characteristics, and induced GBM cell cycle arrest. Overall, this strategy offers a new method for the rapid identification of small-molecule inhibitors of BER enzyme activities that can prevent false negatives due to a fluorescent background.

Keywords

Alkyladenine DNA glycosylase; Drug screening; Glioblastoma; N3-methyladenine; Sunitinib; Temozolomide.

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