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

该子项目是利用该技术的众多研究子项目之一 资源由 NIH/NCRR 资助的中心拨款提供。子项目及 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以在其他 CRISP 条目中表示。列出的机构是 对于中心来说，它不一定是研究者的机构。 AMS 将用于测量铂-DNA 加合物的药物药代动力学。尽管铂类抗癌药物（顺铂、卡铂和奥沙利铂）很重要，但由于传统方法的检测限制，它们的作用机制、受损DNA修复和药代动力学尚不清楚（传统方法无法定量与药理剂量的抗癌药物孵育的细胞中的Pt-DNA加合物，这就是我们需要AMS灵敏度的原因）。为了解决这些重要问题，14C标记的卡铂和奥沙利铂将被用于大肠杆菌、人类细胞和膀胱癌患者，即使在亚药理学剂量下，也可能克服之前的检测极限。目标是使用 AMS 阐明其体内作用机制，使用多种人类癌细胞将 Pt-DNA 加合物水平与细胞死亡相关联，确定服用 14C 标记卡铂和奥沙利铂的患者的药代动力学，并最终将药代动力学结果与个体结果（患者生存）相关联。实验上，在给一些人类癌细胞或癌症患者服用放射性铂基抗癌药物后，细胞裂解并提取铂化DNA将通过AMS进行测量。这些“实时药代动力学”将有助于确定哪些癌症患者将从铂类治疗中受益以及哪些患者会对药物产生耐药性。由于灵敏度高，AMS 是实现这些具有挑战性的目标的最佳技术。

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.