ENT1, a Ribavirin Transporter, Plays a Pivotal Role in Antiviral Efficacy of Ribavirin in a Hepatitis C Virus Replication Cell System

• Published in: Antimicrobial Agents and Chemotherapy Vol. 56; no. 3; pp. 1407 - 1413
• Main Authors: Iikura, Minami, Furihata, Tomomi, Mizuguchi, Misa, Nagai, Miki, Ikeda, Masanori, Kato, Nobuyuki, Tsubota, Akihito, Chiba, Kan
• Format: Journal Article
• Language: English
• Published: Washington, DC American Society for Microbiology 01.03.2012
• Subjects: Antibiotics. Antiinfectious agents. Antiparasitic agents; Antiviral Agents; Antiviral Agents - pharmacology; antiviral properties; Biological and medical sciences; Biological Transport - drug effects; Blotting, Western; Cell Line, Tumor; Equilibrative Nucleoside Transporter 1; Equilibrative Nucleoside Transporter 1 - antagonists & inhibitors; Equilibrative Nucleoside Transporter 1 - metabolism; gene expression; Genes, Reporter; Hepacivirus; Hepacivirus - drug effects; Hepacivirus - physiology; Hepatitis C virus; Hepatocytes; Hepatocytes - drug effects; Hepatocytes - metabolism; Human viral diseases; Humans; Infectious diseases; Luciferases; Medical sciences; messenger RNA; MicroRNAs - genetics; MicroRNAs - metabolism; Pharmacology. Drug treatments; physiological transport; Ribavirin; Ribavirin - pharmacology; RNA, Messenger; RNA, Messenger - genetics; therapeutics; Thioinosine - analogs & derivatives; Thioinosine - pharmacology; transporters; Viral diseases; Viral hepatitis; virus replication; Hepatitis A virus; Picornaviridae; Treatment efficiency; Hepatic disease; Ribavirin; Hepatovirus; Infection; Virus; Viral hepatitis A; Viral disease; Nucleoside analog; Digestive diseases; Antiviral; Replication; Cell system; Flaviviridae; Hepatitis C virus; Hepacivirus; Carrier protein; Viral hepatitis C
• ISSN: 0066-4804; 1098-6596
• DOI: 10.1128/aac.05762-11

We previously showed that equilibrative nucleoside transporter 1 (ENT1) is a primary ribavirin transporter in human hepatocytes. However, because the role of this transporter in the antiviral mechanism of the drug remains unclear, the present study aimed to elucidate the role of ENT1 in ribavirin antiviral action. OR6 cells, a hepatitis C virus (HCV) replication system, were used to evaluate both ribavirin uptake and efficacy. The ribavirin transporter in OR6 cells was identified by mRNA expression analyses and transport assays. Nitrobenzylmercaptopurine riboside (NBMPR) and micro-RNA targeted to ENT1 mRNA (miR-ENT1) were used to reduce the ribavirin uptake level in OR6 cells. Our results showed that ribavirin antiviral activity was associated with its accumulation in OR6 cells, which was also closely associated with the uptake of the drug. It was found that the primary ribavirin transporter in OR6 cells was ENT1 and that inhibition of ENT1-mediated ribavirin uptake by NBMPR significantly attenuated the antiviral activity of the drug as well as its accumulation in OR6 cells. The results also showed that even a small reduction in the ENT1-mediated ribavirin uptake, achieved in this case using miR-ENT1, caused a significant decrease in its antiviral activity, thus indicating that the ENT1-mediated ribavirin uptake level determined its antiviral activity level in OR6 cells. In conclusion, our results show that by facilitating its uptake and accumulation in OR6 cells, ENT1 plays a pivotal role in the antiviral effectiveness of ribavirin and therefore provides an important insight into the efficacy of the drug in anti-HCV therapy.