RT Journal Article
SR Electronic
T1 Enhanced Delivery of Drugs to the Liver by Adenovirus-Mediated Heterologous Expression of the Human Oligopeptide Transporter PEPT1
JF Journal of Pharmacology and Experimental Therapeutics
JO J Pharmacol Exp Ther
FD American Society for Pharmacology and Experimental Therapeutics
SP 812
OP 819
DO 10.1124/jpet.301.3.812
VO 301
IS 3
A1 Toyobuku, Hidekazu
A1 Sai, Yoshimichi
A1 Tamai, Ikumi
A1 Tsuji, Akira
YR 2002
UL http://jpet.aspetjournals.org/content/301/3/812.abstract
AB To explore the feasibility of drug delivery to the liver by the use of adenovirus-mediated human oligopeptide transporter (hPEPT1) gene transfer, we examined the accumulation ofl-[3H]carnosine in the hepatoma cell line (HepG2 and WIFB9) and mouse liver. We constructed a recombinant adenovirus encoding hPEPT1-enhanced yellow fluorescent protein (EYFP) fusion gene (AdhPEPT1-EYFP). In vitro uptake ofl-[3H]carnosine was determined in HepG2 and WIFB9 cells transduced with AdhPEPT1-EYFP. In vivo, the accumulation ofl-[3H]carnosine in mouse liver was evaluated after transduction of AdhPEPT1-EYFP. At pH 6.0, the uptake ofl-[3H]carnosine by HepG2 and WIFB9 cells transduced with AdhPEPT1-EYFP was increased 15- and 2-fold, respectively, compared with the cells without transduction. At pH 7.4, uptake of l-[3H]carnosine in AdhPEPT1-EYFP transduced HepG2 cells was 3 times greater than that of nontransduced cells. In the presence of carnosine or glycylsarcosine as an inhibitor at 20 mM, the uptake of l-[3H]carnosine was reduced to a level comparable to that of nontransduced cells. At 30 min after intravenous administration ofl-[3H]carnosine to mice transduced with AdhPEPT1-EYFP at 1 × 1010 plaque-forming units/mouse, the tissue-to-plasma concentration ratio (Kp) ofl-[3H]carnosine in the liver was significantly increased to 7 times that of nontransduced mice. In contrast, the Kp value of [14C]inulin, a marker for extracellular fluid space, remained unchanged after adenoviral transduction suggesting minimal pathological damage of tissues. hPEPT1-EYFP was localized at both the basolateral and apical membranes in HepG2 cells, WIFB9 cells, and mouse liver. In conclusion, our results suggest that delivery of oligopeptide to the liver by adenovirus-mediated heterologous expression of hPEPT1 in vivo is feasible. The American Society for Pharmacology and Experimental Therapeutics