Functional analysis of single nucleotide polymorphisms of hepatic organic anion transporter OATP1B1 (OATP-C)

Megumi Iwai, Hiroshi Suzuki, Ichiro Ieiri, Kenji Otsubo, Yuichi Sugiyama

Research output: Contribution to journalArticle

131 Citations (Scopus)

Abstract

Objective: Two kinds of single nucleotide polymorphism (SNP; Asn130Asp and Val174Ala) are frequently observed in the liver specific transporter, organic anion transporting polypeptide 1B1 (OATP1B1/OATP-C) gene. Although these two SNPs occur independently in European-Americans, Val174Ala is mostly associated with Asn130Asp in Japanese. Our previous in-vivo studies in Japanese subjects indicated that the non-renal clearance of pravastatin was decreased to 13% of that in wild-type subjects (Nishizato et al. Clin Pharmacol Ther 2003;73(6):554-564). The purpose of the present study is to characterize the function of SNPs variants of OATP1B1 in cDNA transfected cells. Methods: The localization and transport activity were analyzed in HEK293 cells stably expressing wild-type OATP1B1 (OATP1B1*1a), OATP1B1*1b (Asn130Asp), OATP1B1*5 (Val174AIa) and OATP1B1*15 (Asn130Asp and Val174AIa). To characterize the intrinsic Vmax, observed Vmax in uptake study were normalized by the expression level estimated from Western blotting. Results: All SNP variants are predominantly located on the cell surface. No significant alteration was observed in Km values for the transport of 17β-estradiol 17β-D-glucuronide (E217βG), a typical substrate of OATP1B1, among these SNP variants. However, the normalized V max value for OATP1B1*15 was drastically decreased to less than 30% compared with OATP1B1*1a. In contrast, the transport activity of OATP1B1*1 b (Asn130Asp) and OATP1B1*5 (Val 174AIa) was similar to that of OATP1B1*1 a. Conclusions: These results are consistent with the results of our previous clinical studies. It is thus suggested that in-vivo disposition may be predicted from in-vitro results using recombinant transporters.

Original languageEnglish
Pages (from-to)749-757
Number of pages9
JournalPharmacogenetics
Volume14
Issue number11
DOIs
Publication statusPublished - Nov 1 2004
Externally publishedYes

Fingerprint

Organic Anion Transporters
Single Nucleotide Polymorphism
Liver
Pravastatin
HEK293 Cells
Glucuronides
Estradiol
Complementary DNA
Western Blotting
Peptides
Genes

All Science Journal Classification (ASJC) codes

  • Genetics
  • Pharmacology, Toxicology and Pharmaceutics(all)

Cite this

Functional analysis of single nucleotide polymorphisms of hepatic organic anion transporter OATP1B1 (OATP-C). / Iwai, Megumi; Suzuki, Hiroshi; Ieiri, Ichiro; Otsubo, Kenji; Sugiyama, Yuichi.

In: Pharmacogenetics, Vol. 14, No. 11, 01.11.2004, p. 749-757.

Research output: Contribution to journalArticle

Iwai, Megumi ; Suzuki, Hiroshi ; Ieiri, Ichiro ; Otsubo, Kenji ; Sugiyama, Yuichi. / Functional analysis of single nucleotide polymorphisms of hepatic organic anion transporter OATP1B1 (OATP-C). In: Pharmacogenetics. 2004 ; Vol. 14, No. 11. pp. 749-757.
@article{9fccac406b3f404e84684186016224e2,
title = "Functional analysis of single nucleotide polymorphisms of hepatic organic anion transporter OATP1B1 (OATP-C)",
abstract = "Objective: Two kinds of single nucleotide polymorphism (SNP; Asn130Asp and Val174Ala) are frequently observed in the liver specific transporter, organic anion transporting polypeptide 1B1 (OATP1B1/OATP-C) gene. Although these two SNPs occur independently in European-Americans, Val174Ala is mostly associated with Asn130Asp in Japanese. Our previous in-vivo studies in Japanese subjects indicated that the non-renal clearance of pravastatin was decreased to 13{\%} of that in wild-type subjects (Nishizato et al. Clin Pharmacol Ther 2003;73(6):554-564). The purpose of the present study is to characterize the function of SNPs variants of OATP1B1 in cDNA transfected cells. Methods: The localization and transport activity were analyzed in HEK293 cells stably expressing wild-type OATP1B1 (OATP1B1*1a), OATP1B1*1b (Asn130Asp), OATP1B1*5 (Val174AIa) and OATP1B1*15 (Asn130Asp and Val174AIa). To characterize the intrinsic Vmax, observed Vmax in uptake study were normalized by the expression level estimated from Western blotting. Results: All SNP variants are predominantly located on the cell surface. No significant alteration was observed in Km values for the transport of 17β-estradiol 17β-D-glucuronide (E217βG), a typical substrate of OATP1B1, among these SNP variants. However, the normalized V max value for OATP1B1*15 was drastically decreased to less than 30{\%} compared with OATP1B1*1a. In contrast, the transport activity of OATP1B1*1 b (Asn130Asp) and OATP1B1*5 (Val 174AIa) was similar to that of OATP1B1*1 a. Conclusions: These results are consistent with the results of our previous clinical studies. It is thus suggested that in-vivo disposition may be predicted from in-vitro results using recombinant transporters.",
author = "Megumi Iwai and Hiroshi Suzuki and Ichiro Ieiri and Kenji Otsubo and Yuichi Sugiyama",
year = "2004",
month = "11",
day = "1",
doi = "10.1097/00008571-200411000-00006",
language = "English",
volume = "14",
pages = "749--757",
journal = "Pharmacogenetics and Genomics",
issn = "1744-6872",
publisher = "Lippincott Williams and Wilkins",
number = "11",

}

TY - JOUR

T1 - Functional analysis of single nucleotide polymorphisms of hepatic organic anion transporter OATP1B1 (OATP-C)

AU - Iwai, Megumi

AU - Suzuki, Hiroshi

AU - Ieiri, Ichiro

AU - Otsubo, Kenji

AU - Sugiyama, Yuichi

PY - 2004/11/1

Y1 - 2004/11/1

N2 - Objective: Two kinds of single nucleotide polymorphism (SNP; Asn130Asp and Val174Ala) are frequently observed in the liver specific transporter, organic anion transporting polypeptide 1B1 (OATP1B1/OATP-C) gene. Although these two SNPs occur independently in European-Americans, Val174Ala is mostly associated with Asn130Asp in Japanese. Our previous in-vivo studies in Japanese subjects indicated that the non-renal clearance of pravastatin was decreased to 13% of that in wild-type subjects (Nishizato et al. Clin Pharmacol Ther 2003;73(6):554-564). The purpose of the present study is to characterize the function of SNPs variants of OATP1B1 in cDNA transfected cells. Methods: The localization and transport activity were analyzed in HEK293 cells stably expressing wild-type OATP1B1 (OATP1B1*1a), OATP1B1*1b (Asn130Asp), OATP1B1*5 (Val174AIa) and OATP1B1*15 (Asn130Asp and Val174AIa). To characterize the intrinsic Vmax, observed Vmax in uptake study were normalized by the expression level estimated from Western blotting. Results: All SNP variants are predominantly located on the cell surface. No significant alteration was observed in Km values for the transport of 17β-estradiol 17β-D-glucuronide (E217βG), a typical substrate of OATP1B1, among these SNP variants. However, the normalized V max value for OATP1B1*15 was drastically decreased to less than 30% compared with OATP1B1*1a. In contrast, the transport activity of OATP1B1*1 b (Asn130Asp) and OATP1B1*5 (Val 174AIa) was similar to that of OATP1B1*1 a. Conclusions: These results are consistent with the results of our previous clinical studies. It is thus suggested that in-vivo disposition may be predicted from in-vitro results using recombinant transporters.

AB - Objective: Two kinds of single nucleotide polymorphism (SNP; Asn130Asp and Val174Ala) are frequently observed in the liver specific transporter, organic anion transporting polypeptide 1B1 (OATP1B1/OATP-C) gene. Although these two SNPs occur independently in European-Americans, Val174Ala is mostly associated with Asn130Asp in Japanese. Our previous in-vivo studies in Japanese subjects indicated that the non-renal clearance of pravastatin was decreased to 13% of that in wild-type subjects (Nishizato et al. Clin Pharmacol Ther 2003;73(6):554-564). The purpose of the present study is to characterize the function of SNPs variants of OATP1B1 in cDNA transfected cells. Methods: The localization and transport activity were analyzed in HEK293 cells stably expressing wild-type OATP1B1 (OATP1B1*1a), OATP1B1*1b (Asn130Asp), OATP1B1*5 (Val174AIa) and OATP1B1*15 (Asn130Asp and Val174AIa). To characterize the intrinsic Vmax, observed Vmax in uptake study were normalized by the expression level estimated from Western blotting. Results: All SNP variants are predominantly located on the cell surface. No significant alteration was observed in Km values for the transport of 17β-estradiol 17β-D-glucuronide (E217βG), a typical substrate of OATP1B1, among these SNP variants. However, the normalized V max value for OATP1B1*15 was drastically decreased to less than 30% compared with OATP1B1*1a. In contrast, the transport activity of OATP1B1*1 b (Asn130Asp) and OATP1B1*5 (Val 174AIa) was similar to that of OATP1B1*1 a. Conclusions: These results are consistent with the results of our previous clinical studies. It is thus suggested that in-vivo disposition may be predicted from in-vitro results using recombinant transporters.

UR - http://www.scopus.com/inward/record.url?scp=9244254743&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=9244254743&partnerID=8YFLogxK

U2 - 10.1097/00008571-200411000-00006

DO - 10.1097/00008571-200411000-00006

M3 - Article

C2 - 15564882

AN - SCOPUS:9244254743

VL - 14

SP - 749

EP - 757

JO - Pharmacogenetics and Genomics

JF - Pharmacogenetics and Genomics

SN - 1744-6872

IS - 11

ER -