PT  - JOURNAL ARTICLE
AU  - Jennifer H. Gray
AU  - Lara M. Mangravite
AU  - Ryan P. Owen
AU  - Thomas J. Urban
AU  - Wendy Chan
AU  - Elaine J. Carlson
AU  - Conrad C. Huang
AU  - Michiko Kawamoto
AU  - Susan J. Johns
AU  - Douglas Stryke
AU  - Thomas E. Ferrin
AU  - Kathleen M. Giacomini
TI  - Functional and Genetic Diversity in the Concentrative Nucleoside Transporter, CNT1, in Human Populations
AID  - 10.1124/mol.65.3.512
DP  - 2004 Mar 01
TA  - Molecular Pharmacology
PG  - 512--519
VI  - 65
IP  - 3
4099  - http://molpharm.aspetjournals.org/content/65/3/512.short
4100  - http://molpharm.aspetjournals.org/content/65/3/512.full
SO  - Mol Pharmacol2004 Mar 01; 65
AB  - The concentrative nucleoside transporter, CNT1 (SLC28A1), mediates the cellular uptake of naturally occurring pyrimidine nucleosides and many structurally diverse anticancer and antiviral nucleoside analogs. As a first step toward understanding whether genetic variation in CNT1 contributes to variation in the uptake and disposition of clinically used nucleoside analogs, we determined the haplotype structure and functionally analyzed all coding region variants of CNT1 identified in ethnically diverse populations (100 African Americans, 100 European Americans, 30 Asians, 10 Mexican Americans, and 7 Pacific Islanders) (Leabman et al., 2003). A total of 58 coding region haplotypes were identified using PHASE analysis, 44 of which contained at least one amino acid variant. More than half of the coding region haplotypes were population-specific. Using site-directed mutagenesis, 15 protein-altering CNT1 variants, including one amino acid insertion and one base pair (bp) deletion, were constructed and expressed in Xenopus laevis oocytes. All variant transporters took up [3H]thymidine with the exception of CNT1-Ser546Pro, a rare variant, and CNT1–1153del, a single bp deletion found at a frequency of 3% in the African American population. The bp deletion results in a frame-shift followed by a stop-codon. The anticancer nucleoside analog gemcitabine had a reduced affinity for CNT1-Val189Ile (a common CNT1 variant found at a frequency of 26%) compared with reference CNT1 (IC50 = 13.8 ± 0.60 μM for CNT1-reference and 23.3 ± 1.5 μM for CNT1-Val189Ile, p < 0.05). These data suggest that common genetic variants of CNT1 may contribute to variation in systemic and intracellular levels of anti-cancer nucleoside analogs.