MRP2, BSEP, and BCRP Transporter Polymorphisms and Chemotherapy Disposition

MRP2、BSEP 和 BCRP 转运蛋白多态性与化疗处置

• 批准号: 7667453
• 负责人: Richard Hsinshin Ho
• 金额: $ 11.97万
• 依托单位: VANDERBILT UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-08-01 至 2012-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7667453
• 关键词: Adverse effects; Affect; Antineoplastic Agents; Apical; Bile fluid; Binding; Biological Assay; Cancer Patient; Caring; Carrier Proteins; Cell Line; Cell Surface Proteins; Cell membrane; Cell model; Cells; Chemotherapy-Oncologic Procedure; Cholestasis; Clinical Pharmacology; Confocal Microscopy; Cytochromes; Data; Development; Drug Efflux; Drug Kinetics; Drug Prescriptions; Enzymes; Evaluation; Gene Expression; Genetic; Genetic Heterogeneity; Genetic Polymorphism; Genetic Variation; Genotype; Goals; Hand; Hela Cells; Hepatocyte; Immunoblotting; Immunofluorescence Immunologic; In Vitro; Individual; Integral Membrane Protein; Intestines; Kinetics; Knowledge; Liver; Mediating; Membrane; Molecular Biology; Morbidity - disease rate; Organ; Patients; Pharmaceutical Preparations; Pharmacogenetics; Pharmacotherapy; Phase; Play; Principal Investigator; Process; Proteins; Pump; Qualifying; Recombinants; Research Personnel; Research Proposals; Role; Secondary to; Source; Substrate Specificity; Supervision; System; Techniques; Testing; Therapeutic; Therapeutic Index; Toxic effect; Transfection; Vaccinia; Variant; Vesicle; apical membrane; base; bile salts; cancer therapy; chemotherapeutic agent; chemotherapy; clinical efficacy; drug metabolism; experience; improved; in vivo; malignant breast neoplasm; mortality; multidrug resistance-associated protein 2; oncology; programs; protein expression; response; skills; stable cell line

DESCRIPTION (provided by applicant): The proposal seeks the opportunity for the Principal Investigator (PI) to gain knowledge and skills in molecular biology and clinical pharmacology under the direct supervision of two highly qualified sponsors to enhance his potential to develop into a successful independent investigator. The scope of this proposal will incorporate the necessary coursework, seminars, and hands-on experience in new techniques to enable the PI to complete an intensive research proposal over the full five years of the proposal. The overall scientific goal of this proposal is to establish the associations between functional polymorphisms in MRP2, BSEP, and BCRP drug efflux transporters and modulation of chemotherapeutic disposition. Because transporter proteins are critical determinants of the drug disposition process, functional studies in model cell systems will be utilized to evaluate transport activity and perform pharmacokinetic analyses. The hypothesis that transporter variants have significant functional consequences to protein activity and important implications for chemotherapy disposition and toxicity will be evaluated by 2 specific aims: First, MRP2, BCRP, and BSEP variants will be functionally characterized utilizing in vitro recombinant vaccinia-mediated transfection assays in HeLa cells and immunoblot and confocal immunofluoresecent analyses to evaluate total and cell surface protein expression. Second, functionally relevant variants identified in specific aim 1 will be studied in model cell systems utilizing precise functional assays, including inducible gene expression polarized stable cell lines and inside-out membrane vesicles, to provide a comprehensive evaluation of altered mechanisms of function, kinetic analysis, substrate specificity, and chemotherapy-mediated inhibition. Interindividual variation in drug disposition and response is a major concern for commonly prescribed drugs, but has particular relevance with regards to chemotherapeutic agents because efficacy is limited by narrow therapeutic indices. Significant interpatient differences are common with many chemotherapy regimens and it is apparent that genetic factors play vital roles in determining an individual's response to drug therapy. Defining drug transporter genotypes associated with altered disposition of anticancer agents may have significant therapeutic consequences for the use of chemotherapeutic agents in oncology patients by optimizing therapeutic indices and reducing mortality and morbidity associated with adverse effects.

描述（由申请人提供）：该提案为主要研究者（PI）寻求机会，在两个高素质的赞助商的直接监督下，在直接监督分子生物学和临床药理学方面获得知识和技能，以增强其发展为成功的独立研究者的潜力。该提案的范围将在新技术中纳入必要的课程，研讨会和动手经验，以使PI能够在该提案的整整五年内完成一项密集的研究建议。该提案的总体科学目标是建立MRP2，BSEP和BCRP药物外排转运蛋白中功能多态性之间的关联，并调节化学治疗性倾向。由于转运蛋白是药物处置过程的关键决定因素，因此将利用模型细胞系统中的功能研究来评估运输活性并进行药代动力学分析。转运蛋白变体对蛋白质活性具有重大功能后果的假说以及对化学疗法的处置和毒性的重要意义将通过2个具体目的进行评估：第一，MRP2，BCRP和BSEP变体在功能上以功能性地表征了利用内相重组疫苗的转介症和免疫分析的托管分析，并将其概括性化。总和细胞表面蛋白表达。其次，在特定目标1中确定的功能相关变体将在使用精确的功能测定的模型细胞系统中研究，其中包括诱导基因表达极化稳定的稳定细胞系和内部膜囊泡，以全面评估功能改变机制，动力学分析，底物特异性以及化学疗法介导的抑制作用。药物处置和反应的个体差异是普遍规定的药物的主要关注点，但与化学治疗剂有关，因为功效受到狭窄的治疗指数的限制，因此与化学治疗剂有关。许多化学疗法方案很常见，患病室的差异很大，很明显，遗传因素在确定个人对药物治疗的反应中起着至关重要的作用。定义与抗癌药物处置性改变相关的药物转运蛋白基因型，通过优化治疗指数并降低与不良反应相关的死亡率和发病率，可能会对在肿瘤患者中使用化学治疗剂的使用可能产生重大治疗后果。