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）：羰基还原酶活性占了在临床实践中广泛使用的药理学剂的代谢的很大一部分，例如抗精神病药氟哌啶醇和抗癌雌激素雌激素阿霉素和daunorubibicin。在人类中，有两个羰基还原酶，羰基还原酶1（CBR1）和羰基还原酶3（CBR3）。 CBR1和CBR3中的遗传变异可能是CBR底物的药物代谢的广泛变异的关键。将需要一种系统的方法，将常见CBR1和CBH3遗传变异与功能研究结合在一起，以批判性地描绘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。在CBR3中，对蛋白质的氨基酸性序列中非同义变化进行编码的两个SNP被精确地进行了功能表征研究。一个SNP编码valine244对蛋氨酸244的变化（CBR3 V244M），而另一个SNP导致Cysteine4 to Tolosine4 to Tolosine4取代（CBR3 C4Y）。有趣的是，CBR3等位基因在不同种族之间很常见。非常有前途的动力学数据表明，多态性CBR3 Valine244和CBR3蛋白244蛋白质同工型具有与甲二烯和阿霉素的独特催化特性。因此，特定目标1中的研究将集中于多态性CBR3蛋白同工型的功能表征。在特定的目标2中，将使用200个配对的DNA-RNA肝组织样品研究多态性CBR3在肝CBR活性中的影响。基因型 - 表型相关性的存在将通过测量已知CBR3基因型的样品中的酶活性和蛋白质水平等变量来分析。个体间CBR变异性的另一个因素可能取决于调节性CBR1和CBR3近端启动子区域中遗传多态性的存在。在特定的AIM 3中，将筛选来自表型CBR离群值的DNA样品（AIM2中的精确点），以鉴定迄今未开发的CBR1和CBR3近端启动子区域中的新多态性。在确定等位基因频率之后，新型等位基因变体的功能后果将通过使用基因报告基因测定法进行DNA表达分析来检查。总的来说，拟议的研究将提供有关CBR1和CBR3遗传变异对CBR介导的药物代谢的影响的重要信息。对CBR代谢药物的药效学的分子基础的理解将有助于设计更合理的药理疗法。