PHARMACOGENETICS OF HUMAN CARBONYL REDUCTASES

人羧基还原酶的药物遗传学

• 批准号: 7576713
• 负责人: Javier Guillermo Blanco
• 金额: $ 27.05万
• 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 2010-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7576713
• 关键词: Accounting; Alcohol Oxidoreductases; Amino Acid Sequence; Anthracyclines; Antipsychotic Agents; Biological Assay; Classification; Code; Coupled; Cytosol; DNA; Data; Daunorubicin; Doxorubicin; Ethnic group; Gene Frequency; Genes; Genetic; Genetic Polymorphism; Genetic Variation; Genotype; Goals; Haloperidol; Hepatic; Human; Human Genetics; In Vitro; Individual; Isotopically-Coded Affinity Tagging; Kinetics; Liquid Chromatography; Liver; Mass Spectrum Analysis; Measures; Mediating; Messenger RNA; Metabolic Biotransformation; Metabolism; Molecular; Peptide Sequence Determination; Persons; Pharmaceutical Preparations; Pharmacodynamics; Pharmacogenetics; Phenotype; Play; Promoter Regions; Property; Protein Isoforms; Proteins; Reporter Genes; Research; Role; Sampling; Single Nucleotide Polymorphism; Time; Tissue Sample; Variant; Vitamin K 3; Western Blotting; base; clinical practice; design; drug metabolism; enzyme activity; genetic variant; in vitro Model; in vivo; novel; response

DESCRIPTION (PROVIDED BY APPLICANT): Carbonyl reductase activity accounts for a significant fraction of the metabolism of pharmacological agents extensively used in clinical practice such as the antipsychotic haloperidol and the anticancer anthracyclines doxorubicin and daunorubicin. In humans there are two carbonyl reductases, carbonyl reductase 1 (CBR1) and carbonyl reductase 3 (CBR3). It is possible that genetic variability in CBR1 and CBR3 may be key for the wide person-to-person variation in the metabolism of drugs that are CBR substrates. A systematic approach that combines the identification of common CBR1 and CBH3 genetic variants together with functional studies will be needed to critically delineate the role of CBR1 and CBR3 in variable CBR mediated drug biotransformation. Towards this goal we investigated the presence of single nucleotide polymorphisms (SNPs) in CBR1 and CBR3. Two SNPs in CBR3 encoding for non-synonymous changes in the amino acidic sequence of the protein were pinpointed to perform functional characterization studies. One SNP encodes for a valine244 to methionine244 change (CBR3 V244M), while the other SNP results in a cysteine4 to tyrosine4 substitution (CBR3 C4Y). Interestingly, the CBR3 allelic variants are common among different ethnic groups. Very promising kinetic data suggest that the polymorphic CBR3 valine244 and CBR3 methionine244 protein isoforms have distinctive catalytic properties towards menadione and doxorubicin. Thus, studies in specific Aim 1 will focus on the functional characterization of the polymorphic CBR3 protein isoforms. In specific Aim 2 the effects of polymorphic CBR3 in hepatic CBR activity will be investigated using 200 paired DNA-RNA liver tissue samples. The presence of genotype-phenotype correlations will be analyzed by measuring variables such as enzyme activities and protein levels in samples with known CBR3 genotypes. An additional contributing factor for inter-individual CBR variability may be dictated by the presence of genetic polymorphisms in the regulatory CBR1 and CBR3 proximal promoter regions. In specific Aim 3, DNA samples from phenotypic CBR outliers (pinpointed in Aim2) will be screened to identify new polymorphisms in the thus far unexplored CBR1 and CBR3 proximal promoter regions. Subsequent to the determination of allele frequencies, the functional consequences of the novel allelic variants will be examined by DNA expression analysis using gene reporter assays. Collectively, the proposed studies will provide essential information on the impact of CBR1 and CBR3 genetic variability on CBR mediated drug metabolism. The understanding of the molecular basis that govern the pharmacodynamics of CBR metabolized drugs will assist the design of more rational pharmacological therapies.

描述（由申请人提供）：羰基还原酶活性占临床实践中广泛使用的药物代谢的重要部分，例如抗精神病药氟哌啶醇和抗癌蒽环类药物阿霉素和柔红霉素。人类有两种羰基还原酶：羰基还原酶 1 (CBR1) 和羰基还原酶 3 (CBR3)。 CBR1 和 CBR3 的遗传变异可能是 CBR 底物药物代谢存在广泛的人际差异的关键。需要一种将常见 CBR1 和 CBH3 遗传变异的鉴定与功能研究相结合的系统方法，以批判性地描述 CBR1 和 CBR3 在可变 CBR 介导的药物生物转化中的作用。为了实现这一目标，我们研究了 CBR1 和 CBR3 中单核苷酸多态性 (SNP) 的存在。确定了 CBR3 中编码蛋白质氨基酸序列非同义变化的两个 SNP，以进行功能表征研究。一个 SNP 编码缬氨酸 244 到蛋氨酸 244 的变化 (CBR3 V244M)，而另一个 SNP 导致半胱氨酸 4 到酪氨酸 4 的取代 (CBR3 C4Y)。有趣的是，CBR3 等位基因变异在不同种族群体中很常见。非常有前景的动力学数据表明，多态性 CBR3 缬氨酸 244 和 CBR3 蛋氨酸 244 蛋白亚型对甲萘醌和阿霉素具有独特的催化特性。因此，具体目标 1 的研究将集中于多态性 CBR3 蛋白亚型的功能表征。在具体目标 2 中，将使用 200 个配对的 DNA-RNA 肝脏组织样本研究多态性 CBR3 对肝脏 CBR 活性的影响。将通过测量具有已知 CBR3 基因型的样品中的酶活性和蛋白质水平等变量来分析基因型-表型相关性的存在。个体间 CBR 变异的另一个影响因素可能是由调节性 CBR1 和 CBR3 近端启动子区域中遗传多态性的存在决定的。在特定目标 3 中，将筛选来自表型 CBR 异常值（在目标 2 中精确定位）的 DNA 样本，以识别迄今为止尚未探索的 CBR1 和 CBR3 近端启动子区域中的新多态性。确定等位基因频率后，将使用基因报告分析通过 DNA 表达分析来检查新型等位基因变体的功能后果。总的来说，拟议的研究将提供有关 CBR1 和 CBR3 遗传变异对 CBR 介导的药物代谢影响的重要信息。了解控制 CBR 代谢药物药效学的分子基础将有助于设计更合理的药理学疗法。