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

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 AMS将用于药物药代动力学的铂-DNA加合物的测定。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极限，它们的作用机制、DNA损伤修复和药代动力学尚不清楚(传统方法无法用药理剂量的抗癌剂孵育细胞来定量铂-DNA加合物，这就是为什么我们需要AMS的灵敏度)。为了解决这些重要问题，14C标记的卡铂和奥沙利铂将用于大肠杆菌、人类细胞和膀胱癌患者，即使在亚药物剂量下也可能克服以前的检测限值。我们的目标是利用AMS阐明它们在体内的作用机制，利用各种人类癌细胞将铂-DNA加合物水平与细胞死亡联系起来，确定服用14C标记的卡铂和奥沙利铂患者的药代动力学，并最终将药代动力学结果与个体结果(患者生存)相关联。在实验上，在给一些人类癌细胞或癌症患者服用以铂为基础的放射性抗癌药物后，细胞裂解和提取的铂DNA将被AMS测量。这些“实时药代动力学”将能够确定哪些癌症患者将受益于铂类药物治疗，哪些患者将对药物产生抗药性。由于其高度的敏感性，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.