Departments of Pharmaceutics (D.K.B., A.B., H.Z., K.G.C., A.M., B.P.), Genome Sciences (S.L., D.A.N.), Biostatistics (T.A.T.), and Medicine (J.K.A.), University of Washington, Seattle, Washington; Division of Pediatric Pharmacology and Medical Toxicology, Department of Pediatrics, Children's Mercy Hospitals and Clinics, Kansas City, Missouri (A.G., R.E.P., R.G., J.S.L.); Department of Pharmaceutical Sciences, St. Jude Children's Research Hospital, Memphis, Tennessee (A.S.C., E.G.S.); and Section of Genomic Pediatrics, Department of Pediatrics, and Human and Molecular Genetics Center, Medical College of Wisconsin, Milwaukee, Wisconsin (U.B.).
Departments of Pharmaceutics (D.K.B., A.B., H.Z., K.G.C., A.M., B.P.), Genome Sciences (S.L., D.A.N.), Biostatistics (T.A.T.), and Medicine (J.K.A.), University of Washington, Seattle, Washington; Division of Pediatric Pharmacology and Medical Toxicology, Department of Pediatrics, Children's Mercy Hospitals and Clinics, Kansas City, Missouri (A.G., R.E.P., R.G., J.S.L.); Department of Pharmaceutical Sciences, St. Jude Children's Research Hospital, Memphis, Tennessee (A.S.C., E.G.S.); and Section of Genomic Pediatrics, Department of Pediatrics, and Human and Molecular Genetics Center, Medical College of Wisconsin, Milwaukee, Wisconsin (U.B.)
Drug Metab Dispos. 2018 Jun;46(6):888-896. doi: 10.1124/dmd.118.080952. Epub 2018 Mar 30.
The major objective of this study was to investigate the association of genetic and nongenetic factors with variability in protein abundance and in vitro activity of the androgen-metabolizing enzyme UGT2B17 in human liver microsomes ( = 455). UGT2B17 abundance was quantified by liquid chromatography-tandem mass spectrometry proteomics, and enzyme activity was determined by using testosterone and dihydrotestosterone as in vitro probe substrates. Genotyping or gene resequencing and mRNA expression were also evaluated. Multivariate analysis was used to test the association of UGT2B17 copy number variation, single nucleotide polymorphisms (SNPs), age, and sex with its mRNA expression, abundance, and activity. UGT2B17 gene copy number and SNPs (rs7436962, rs9996186, rs28374627, and rs4860305) were associated with gene expression, protein levels, and androgen glucuronidation rates in a gene dose-dependent manner. UGT2B17 protein (mean ± S.D. picomoles per milligram of microsomal protein) is sparsely expressed in children younger than 9 years (0.12 ± 0.24 years) but profoundly increases from age 9 years to adults (∼10-fold) with ∼2.6-fold greater abundance in males than in females (1.2 vs. 0.47). Association of androgen glucuronidation with UGT2B15 abundance was observed only in the low UGT2B17 expressers. These data can be used to predict variability in the metabolism of UGT2B17 substrates. Drug companies should include UGT2B17 in early phenotyping assays during drug discovery to avoid late clinical failures.
本研究的主要目的是研究遗传和非遗传因素与雄激素代谢酶 UGT2B17 在人肝微粒体中的蛋白丰度和体外活性变异性的关系（=455）。通过液相色谱-串联质谱蛋白质组学定量 UGT2B17 的丰度，并使用睾酮和二氢睾酮作为体外探针底物测定酶活性。还评估了基因分型或基因重测序和 mRNA 表达。多变量分析用于检验 UGT2B17 拷贝数变异、单核苷酸多态性（SNPs）、年龄和性别与 mRNA 表达、丰度和活性的关系。UGT2B17 基因拷贝数和 SNPs（rs7436962、rs9996186、rs28374627 和 rs4860305）与基因表达、蛋白水平和雄激素葡萄糖醛酸化率呈基因剂量依赖性相关。UGT2B17 蛋白（平均值±S.D. 皮摩尔/毫克微粒体蛋白）在 9 岁以下儿童中表达稀疏（0.12±0.24 岁），但从 9 岁到成年后显著增加（约 10 倍），男性比女性多约 2.6 倍（1.2 比 0.47）。仅在低表达 UGT2B17 的个体中观察到雄激素葡萄糖醛酸化与 UGT2B15 丰度的关联。这些数据可用于预测 UGT2B17 底物代谢的变异性。制药公司应在药物发现早期表型分析中纳入 UGT2B17，以避免后期临床失败。
