Pharmacogenetics of Human Carbonyl Reductases

人羰基还原酶的药物遗传学

• 批准号: 7982748
• 负责人: Javier Guillermo Blanco
• 金额: $ 31.7万
• 依托单位: STATE UNIVERSITY OF NEW YORK AT BUFFALO
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-03-01 至 2014-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7982748
• 关键词: 2-tert-butylhydroquinone; 3' Untranslated Regions; Affect; Alcohol Oxidoreductases; Alcohols; Anthracyclines; Antioxidants; Antipsychotic Agents; Area; Binding; Biological Assay; Cardiotoxicity; Child; Clinical; Complex; Correlation Studies; Coupled; DNA Sequence; Data; Daunorubicin; Development; Doxorubicin; Drug toxicity; Duborimycin; Electrons; Flavonoids; Gene Expression Regulation; Genes; Genetic Determinism; Genetic Polymorphism; Genetic Variation; Genotype; Goals; Haloperidol; Heart; Hepatic Tissue; Human; Human Genetics; Immunoblotting; Individual; Investigation; Knowledge; Liquid Chromatography; Malignant Neoplasms; Mass Spectrum Analysis; Matched Case-Control Study; Mediating; Metabolism; MicroRNAs; Molecular; Mono-S; Myocardial tissue; Myocardium; National Institute of General Medical Sciences; Oxidation-Reduction; Pathogenesis; Pharmaceutical Preparations; Pharmacodynamics; Pharmacogenetics; Phase II Clinical Trials; Phenotype; Play; Production; Promoter Regions; Property; Protein Isoforms; Proteins; Regulation; Relative (related person); Reporting; Research; Response Elements; Risk; Role; Rutin; Sampling; Single Nucleotide Polymorphism; Stimulus; Surveys; Testing; Thermodynamics; Time; Tissue Sample; Tissues; Variant; Vitamin K 3; base; cancer therapy; childhood cancer survivor; clinical practice; clinically relevant; design; enzyme activity; genetic risk factor; in vitro Model; in vivo; inhibitor/antagonist; interest; mRNA Expression; nano; nuclear factor-erythroid 2; pre-clinical; promoter; protein expression; public health relevance; research study; response; transcription factor; validation studies

DESCRIPTION (provided by applicant): Human carbonyl reductases (CBRs) catalyze the reduction of several drugs widely used in clinical practice including the anticancer anthracyclines doxorubicin and daunorubicin. The pharmacodynamics of these 2 drugs is unpredictable. We hypothesize that interindividual variability in CBR activity contributes to the unpredictable pharmacodynamic profiles for doxorubicin and daunorubicin. Therefore, our main goal is to characterize the molecular basis of variable CBR activity as a prerequisite for the design of more effective anticancer therapies. Thus far, we have (1) characterized functional allelic variants of carbonyl reductase 1 (CBR1) and carbonyl reductase 3 (CBR3); (2) documented the variability in CBR1 and CBR3 expression in hepatic tissue; and (3) identified a genetic risk factor (CBR3 V244M) for anthracycline-related cardiotoxicity in pediatric cancer survivors. Gene regulation studies indicate that specific DNA sequences in the promoter regions of CBR1 and CBR3 influence the level of protein expression and activity in response to various stimuli. New data indicate that CBR3 mRNA expression increases considerably (8.5-fold) in the presence of the prototypical antioxidant tert-butylhydroquinone and that the CBR3 promoter contains 2 conserved antioxidant response elements (AREs). We have designed experiments that will allow us to characterize the functional role of AREs in the induction of CBR3 expression in response to antioxidant exposure (Specific Aim 1). These experiments will also allow us to determine how 2 common CBR3 promoter polymorphisms (CBR3 -725T>C, and CBR3 -326T>A) modulate gene promoter activity in response to antioxidants. A common CBR1 polymorphism (1096G>A) dictates the synthesis of cardiotoxic doxorubicinol in human hepatic tissue. We have planned experiments to determine whether the effect of CBR1 1096G>A is mediated through the binding of specific microRNAs to the polymorphic 3'-untranslated region (Specific Aim 2). A growing amount of experimental evidence, together with our pharmacogenetic findings, suggests that CBR1 and CBR3 have a crucial role in the complex pharmacodynamics of anthracyclines in the heart. The expression of CBR1 and CBR3 in the human heart has not been characterized. We plan to document the relative contributions of CBR1 and CBR3 to the metabolism of doxorubicin and daunorubicin in 200 samples of human myocardial tissue (Specific Aim 3). In this comprehensive approach we will use quantitative real-time PCR analysis, nano-liquid chromatography coupled to triple quadruple mass spectroscopy, and enzyme activity assays with CBR substrates (e.g., doxorubicin) and inhibitors (e.g., the cardioprotective flavonoid monohydroxyethyl rutoside, or mono-HER). We will also conduct genotype-phenotype correlation studies to determine whether functional CBR1 and CBR3 polymorphisms affect the formation of cardiotoxic anthracycline metabolites in the heart. The body of knowledge gathered from the proposed research will contribute to the development of anticancer therapy that can be individualized by identifying the genetic determinants of variable CBR activity. PUBLIC HEALTH RELEVANCE: Human carbonyl reductases (CBR1 and CBR3) catalyze the reduction of several drugs including the anticancer anthracyclines doxorubicin and daunorubicin. The anthracycline alcohol metabolites synthesized by CBR activity are cardiotoxic. Three research aims will investigate (1) the regulation of polymorphic CBR3, (2) the functional impact of a common polymorphism in human CBR1, and (3) the pharmacogenetics of CBR1 and CBR3 with doxorubicin and daunorubicin in human myocardium, the target tissue for anthracycline-related cardiotoxicity.

描述（由申请人提供）：人羰基还原酶（CBR）催化临床实践中广泛使用的几种药物的还原，包括抗癌蒽环类药物多柔比星和柔红霉素。这两种药物的药效学是不可预测的。我们假设CBR活性的个体间变异性导致了阿霉素和柔红霉素不可预测的药效学特征。因此，我们的主要目标是表征可变CBR活性的分子基础，作为设计更有效的抗癌疗法的先决条件。到目前为止，我们已经（1）表征了羰基还原酶1（CBR 1）和羰基还原酶3（CBR 3）的功能等位基因变体;（2）记录了肝组织中CBR 1和CBR 3表达的变异性;（3）确定了儿科癌症幸存者中蒽环类药物相关心脏毒性的遗传风险因素（CBR 3 V244 M）。基因调控研究表明，在CBR 1和CBR 3的启动子区域的特定DNA序列的影响蛋白质的表达水平和活性，在响应各种刺激。新的数据表明，CBR 3 mRNA的表达显着增加（8.5倍）的原型抗氧化剂叔丁基对苯二酚的存在下，CBR 3启动子包含2个保守的抗氧化反应元件（战神）。我们设计的实验将使我们能够表征战神在诱导CBR 3表达中的功能作用，以响应抗氧化剂暴露（具体目标1）。这些实验还将使我们能够确定2种常见的CBR 3启动子多态性（CBR 3 - 725 T>C和CBR 3 - 326 T>A）如何响应抗氧化剂调节基因启动子活性。一种常见的CBR 1多态性（1096 G>A）决定了心脏毒性阿霉素酚在人肝组织中的合成。我们已经计划了实验来确定CBR 11096 G>A的作用是否是通过特异性microRNA与多态性3 '-非翻译区（特异性目标2）的结合来介导的。越来越多的实验证据以及我们的药物遗传学发现表明，CBR 1和CBR 3在心脏中蒽环类药物的复杂药效学中起着至关重要的作用。CBR 1和CBR 3在人类心脏中的表达尚未被表征。我们计划在200份人类心肌组织样本中记录CBR 1和CBR 3对阿霉素和柔红霉素代谢的相对贡献（特定目标3）。在这种全面的方法中，我们将使用定量实时PCR分析，纳米液相色谱法耦合到三重四重质谱法，以及CBR底物的酶活性测定（例如，阿霉素）和抑制剂（例如，心脏保护类黄酮单羟乙基芸香苷，或单HER）。我们还将进行基因型-表型相关性研究，以确定功能性CBR 1和CBR 3多态性是否影响心脏毒性蒽环类代谢物的形成。从拟议的研究中收集的知识体系将有助于抗癌治疗的发展，可以通过识别可变CBR活性的遗传决定因素来个性化。 公共卫生关系：人羰基还原酶（CBR 1和CBR 3）催化几种药物的还原，包括抗癌蒽环类药物阿霉素和柔红霉素。由CBR活性合成的蒽环类醇代谢产物具有心脏毒性。三个研究目标将调查（1）多态性CBR 3的调节，（2）人类CBR 1中常见多态性的功能影响，以及（3）CBR 1和CBR 3与阿霉素和柔红霉素在人类心肌中的药物遗传学，蒽环类药物相关心脏毒性的靶组织。