DEVELOP ASSAY TO QUANT PLAT-DNA ADDUCTS & PREDICT RESPONSE TO CHEMOTHERA

该子项目是利用 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。列出的机构是 中心，不一定是研究者的机构。 AMS将用于铂-DNA加合物的药物动力学研究。尽管基于铂的抗癌药物（顺铂、卡铂和奥沙利铂）很重要，但由于常规方法的检测限，它们的作用机制、受损DNA的修复和药代动力学尚不清楚（常规方法无法定量与药理剂量的抗癌剂孵育的细胞的Pt-DNA加合物，这是我们需要AMS灵敏度的原因）。为了解决这些重要问题，将对E.大肠杆菌，人类细胞和膀胱癌患者，这可能会克服以前的检测极限，甚至在亚药理剂量。目标是使用AMS阐明其体内作用机制，使用各种人类癌细胞将Pt-DNA加合物水平与细胞死亡相关联，确定14 C标记卡铂和奥沙利铂给药患者的药代动力学，并最终将药代动力学结果与个体结局（患者生存期）相关联。在实验上，在用放射性铂基抗癌药物给药许多人类癌细胞或癌症患者后，将通过AMS测量细胞裂解和提取的铂化DNA。这些“实时药代动力学”将允许确定哪些癌症患者将从铂治疗中受益，哪些将对药物产生耐药性。由于灵敏度高，AMS是实现这些挑战性目标的最佳技术。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. AMS will be applied to measuring the platinum-DNA adducts for drug pharmacokinetics. In spite of the importance of platinum-based anticancer drugs (cisplatin, carboplatin and oxaliplatin), their mechanisms of action, repair of damaged DNA and pharmacokinetics are unclear because of the detection limit of conventional methods (conventional methods have failed in quantifying Pt-DNA adducts with cells incubated with a pharmacological dose of the anticancer agent, which is the reason we need the sensitivity of AMS). In order to address these important issues, 14C-labeled carboplatin and oxaliplatin will be administered to E. coli, human cells, and bladder cancer patients, which may overcome the previous detection limits even at sub-pharmacological doses. The goals are to use AMS to elucidate their in vivo mechanism of action, to correlate Pt-DNA adduct level with cell death using a variety of human cancer cells, to determine the pharmacokinetics of the patients dosed with 14C-labeled carboplatin and oxaliplatin, and to ultimately correlate the phamacokinetic results to individual outcome (patient survival). Experimentally, after dosing a number of human cancer cells or cancer patients with radioactive platinum-based anticancer drugs, cell lysis and extracted platinated DNA will be measured by AMS. These 'real-time pharmacokinetics' will allow determination of which cancer patients will benefit from platinum treatment and which will be resistant to the drugs. Because of the high sensitivity, AMS is the very best technology for realizing these challenging goals.