Systematic Screening of Human ABCC3 Polymorphisms and Their Effects on MRP3 Expression and Function

• Published in: Drug metabolism and pharmacokinetics 2011-01, Vol.26 (4), p.374-386
• Main Authors: Sasaki, Tomohiro, Hirota, Takeshi, Ryokai, Yuriko, Kobayashi, Daisuke, Kimura, Miyuki, Irie, Shin, Higuchi, Shun, Ieiri, Ichiro
• Format: Article
• Language: English
• Subjects: ABCC3; Adult; African Americans - genetics; Asian Americans - genetics; DNA, Complementary; drug transporter; European Continental Ancestry Group - genetics; Female; Gene Amplification; Genes, Reporter; Genotype; Haplotypes; human; Humans; Hymecromone - analogs & derivatives; Hymecromone - blood; Hymecromone - metabolism; Hymecromone - pharmacokinetics; Hymecromone - urine; Luciferases - genetics; Luciferases - metabolism; Male; MRP3; Multidrug Resistance-Associated Proteins - biosynthesis; Multidrug Resistance-Associated Proteins - genetics; Multidrug Resistance-Associated Proteins - metabolism; pharmacogenomics; pharmacokinetics; polymorphism; Polymorphism, Genetic; RNA, Messenger - genetics; RNA, Messenger - metabolism; Transfection; Young Adult
• ISSN: 1347-4367; 1880-0920
• DOI: 10.2133/dmpk.DMPK-10-RG-103

The present study was undertaken to identify genetic polymorphisms of multidrug resistance-associated protein 3 (MRP3, gene name ABCC3), an ATP-binding cassette transporter that mediates the transport of substrates across the basolateral membrane into the blood, and to investigate their effects on ABCC3 expression and MRP3 function. We identified genetic polymorphisms of ABCC3 and evaluated the effects by (1) a luciferase reporter gene assay, (2) measuring mRNA levels, and (3) a human pharmacogenomics study with 4-methylumbelliferone glucuronide (4-MUG). Overall, 61 genetic variants were identified in three ethnic populations; of these variants 17 were novel (7 were non-synonymous: 61Arg>Cys, 132Gln>Stop, 221Trp>Stop, 270His>Gln, 548Leu>Gln, 600Lys>Arg, and 1324Arg> His). However, these mutations occurred at very low frequencies (max. 4.7%). The observed allele frequencies showed considerable inter-ethnic differences. The reporter gene assay indicated a significant reduction of transcriptional activity with the –1767G>A allele compared to the wild-type allele; however, a decreased expression of ABCC3 mRNA was not detected in human liver samples. A human pharmacokinetic study showed that the ABCC3 genotype in the promoter region was not associated with changes in the pharmacokinetics of 4-MUG, a substrate of MRP3. This is the first study to assess the effects of ABCC3 polymorphisms on human pharmacokinetics; however, further investigations are needed to complete the picture.