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

作　　者：Ying-Qi Song Guo-Dong Li Dou Niu Feng Chen Shaozhen Jing Vincent Kam Wai Wong Wanhe Wang Chung-Hang Leung 

机构地区：[1]State Key Laboratory of Quality Research in Chinese Medicine,Institute of Chinese Medical Sciences,University of Macao,Macao,999078,China [2]Department of Biomedical Sciences,Faculty of Health Sciences,University of Macao,Macao,999078,China [3]Institute of Medical Research,Northwestern Polytechnical University,Xi'an,710072,China [4]Dr.Neher's Biophysics Laboratory for Innovative Drug Discovery,State Key Laboratory of Quality Research in Chinese Medicine,Macao University of Science and Technology,Macao,999078,China

出　　处：《Journal of Pharmaceutical Analysis》2023年第5期514-522,共9页药物分析学报（英文版）

基　　金：supported by the Science and Technology Development Fund(Grant Nos.:0007/2020/A1 and 0020/2022/A1);the State Key Laboratory of Quality Research in Chinese Medicine,University of Macao(Grant No.:SKL-QRCM(UM)-2020-2022);the University of Macao(Grant Nos.:MYRG2019-00002-ICMS and MYRG2020-00017-ICMS);2022 Internal Research Grant of SKLQRCM(University of Macao)(Grant No.:QRCM-IRG2022-011);the National Natural Science Foundation of China(Grant No.:22101230);the Natural Science Basic Research Program of Shaanxi(Grant No.:2021JQ-089);the Natural Science Foundation of Chongqing,China(Grant No.:cstc2021jcyj-msxmX0659).

摘　　要：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.

关 键 词：Drug screening Alkyladenine DNA glycosylase N3-methyladenine GLIOBLASTOMA TEMOZOLOMIDE SUNITINIB 

分 类 号：R91[医药卫生—药学]