title: Direct detection and quantification of abasic sites for in vivo studies of DNA damage and repair.
abstract: Use of chemotherapeutic agents to induce cytotoxic DNA damage and programmed cell death is a key strategy in cancer treatments. However, the efficacy of DNA-targeted agents such as temozolomide is often compromised by intrinsic cellular responses such as DNA base excision repair (BER). Previous studies have shown that BER pathway resulted in formation of abasic or apurinic/apyrimidinic (AP) sites, and blockage of AP sites led to a significant enhancement of drug sensitivity due to reduction of DNA base excision repair. Since a number of chemotherapeutic agents also induce formation of AP sites, monitoring of these sites as a clinical correlate of drug effect will provide a useful tool in the development of DNA-targeted chemotherapies aimed at blocking abasic sites from repair. Here we report an imaging technique based on positron emission tomography (PET) that allows for direct quantification of AP sites in vivo. For this purpose, positron-emitting carbon-11 has been incorporated into methoxyamine ([(11)C]MX) that binds covalently to AP sites with high specificity. The binding specificity of [(11)C]MX for AP sites was demonstrated by in vivo blocking experiments. Using [(11)C]MX as a radiotracer, animal PET studies have been conducted in melanoma and glioma xenografts for quantification of AP sites. Following induction of AP sites by temozolomide, both tumor models showed significant increase of [(11)C]MX uptake in tumor regions in terms of radioactivity concentration as a function of time, which correlates well with conventional aldehyde reactive probe (ARP)-based bioassays for AP sites.
authors.affiliation: Division of Radiopharmaceutical Science, Case Center for Imaging Research, Department of Radiology, Case Western Reserve University, Cleveland, OH 44122, USA. yanming.wang@case.edu
authors.forename: Yanming
authors.initials: Y
authors.lastname: Wang
authors.forename: Lili
authors.initials: L
authors.lastname: Liu
authors.forename: Chunying
authors.initials: C
authors.lastname: Wu
authors.forename: Alina
authors.initials: A
authors.lastname: Bulgar
authors.forename: Eduardo
authors.initials: E
authors.lastname: Somoza
authors.forename: Wenxia
authors.initials: W
authors.lastname: Zhu
authors.forename: Stanton L
authors.initials: SL
authors.lastname: Gerson
mesh_terms: D000818:Animals; D001483:Base Sequence; D002250:Carbon Radioisotopes / Q000493:pharmacokinetics; D045744:Cell Line, Tumor; D004249:DNA Damage / Q000235:genetics*; D004260:DNA Repair / Q000235:genetics*; D005260:Female; D005910:Glioma / Q000000981:diagnostic imaging* / Q000235:genetics*; D006898:Hydroxylamines / Q000493:pharmacokinetics*; D008545:Melanoma / Q000000981:diagnostic imaging* / Q000235:genetics*; D051379:Mice; D008819:Mice, Nude; D008969:Molecular Sequence Data; D011877:Radionuclide Imaging; D019275:Radiopharmaceuticals / Q000493:pharmacokinetics
chemical_list: D002250:Carbon Radioisotopes; D006898:Hydroxylamines; D019275:Radiopharmaceuticals; C005214:methoxyamine