The human organic anion transporter 3 (OAT3; SLC22A8):: genetic variation and functional genomics
The human organic anion transporter 3 (OAT3; SLC22A8):: genetic variation and functional genomics
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DOI：
10.1152/ajprenal.00272.2005
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发表时间：
2006-04-01
影响因子：
4.2
通讯作者：
Giacomini, KM
中科院分区：
文献类型：
--
作者：
Erdman, AR;Mangravite, LM;Giacomini, KM
人有机阴离子转运蛋白 OAT3 (SLC22A8) 通过介导多种有机阴离子（包括许多常用药物）进入肾近端肾小管细胞，在肾脏药物消除中发挥关键作用。为了了解 OAT3 遗传变异的性质和程度，并确定这种变异是否影响其功能，我们在大量不同种族的样本群体中鉴定了 OAT3 变异，并在细胞测定中研究了它们的转运活性。我们在 270 名个体（80 名非洲裔美国人、80 名欧洲裔美国人、60 名亚裔美国人和 50 名墨西哥裔美国人）的 DNA 样本中总共鉴定出了 10 个不同的编码区变异，这些变异改变了编码的氨基酸序列。这些 OAT3 变体的总体患病率在筛查人群中相对较低，只有 3 个变体在特定种族中的等位基因频率 > 1%。通过定点诱变创建每个变体的克隆，在 HEK-293 细胞中表达，并使用模型底物硫酸雌酮 (ES) 和西咪替丁 (CIM) 测试其功能。结果显示，OAT3 变体之间存在高度的功能异质性，其中三种变体（p.Arg149Ser、p.Gln239Stop 和 p.Ile260Arg）导致功能完全丧失，而其他几个变体则功能显着降低。在 3.5% 的亚裔美国人中发现了一种更常见的变体 (p. Ile305Phe)，它似乎改变了底物特异性。该变体表现出运输 ES 的能力降低，但运输 CIM 的能力保留。这些数据表明 OAT3 的遗传变异可能导致药物处置的变化。
The human organic anion transporter, OAT3 ( SLC22A8), plays a critical role in renal drug elimination, by mediating the entry of a wide variety of organic anions, including a number of commonly used pharmaceuticals, into the renal proximal tubular cells. To understand the nature and extent of genetic variation in OAT3, and to determine whether such variation affects its function, we identified OAT3 variants in a large, ethnically diverse sample population and studied their transport activities in cellular assays. We identified a total of 10 distinct coding-region variants, which altered the encoded amino acid sequence, in DNA samples from 270 individuals ( 80 African-Americans, 80 European-Americans, 60 Asian-Americans, and 50 Mexican-Americans). The overall prevalence of these OAT3 variants was relatively low among the screened population, with only three variants having allele frequencies of > 1% in a particular ethnic group. Clones of each variant were created by site-directed mutagenesis, expressed in HEK-293 cells, and tested for function using the model substrates, estrone sulfate ( ES) and cimetidine ( CIM). The results revealed a high degree of functional heterogeneity among OAT3 variants, with three variants ( p. Arg149Ser, p. Gln239Stop, and p. Ile260Arg) that resulted in complete loss of function, and several others with significantly reduced function. One of the more common variants ( p. Ile305Phe), found in 3.5% of Asian-Americans, appeared to have altered substrate specificity. This variant exhibited a reduced ability to transport ES, but a preserved ability to transport CIM. These data suggest that genetic variation in OAT3 may contribute to variation in the disposition of drugs.