Inhibition of hepatic organic anion-transporting polypeptide by RNA interference in sandwich-cultured human hepatocytes: an in vitro model to assess transporter-mediated drug-drug interactions.
Organic anion-transporting polypeptides (OATPs), members of the SLCO/SLC21 family, mediate the transport of various endo- and xenobiotics. In human liver, OATP1B1, 1B3, and 2B1 are located at the basolateral membrane of hepatocytes and are involved in hepatic drug uptake and biliary elimination. Clinically significant drug-drug interactions (DDIs) mediated by hepatic OATPs have drawn great attention from clinical practitioners and researchers. However, there are considerable challenges to prospectively understanding the extent of OATP-mediated DDIs because of the lack of specific OATP inhibitors or substrates and the limitations of in vitro tools. In the present study, a novel RNA interference knockdown sandwich-cultured human hepatocyte model was developed and validated. Quantitative polymerase chain reaction, microarray and immunoblotting analyses, along with uptake assays, illustrated that the expression and transport activity of hepatic OATPs were reduced by small interfering (siRNA) efficiently and specifically in this model. Although OATP siRNA decreased only 20 to 30% of the total uptake of cerivastatin into human hepatocytes, it caused a 50% reduction in cerivastatin metabolism, which was observed by monitoring the formation of the two major metabolites of cerivastatin. The results suggest that coadministration of a drug that is a hepatic OATP inhibitor could significantly alter the pharmacokinetic profile of cerivastatin in clinical studies. Further studies with this novel model demonstrated that OATP and cytochrome P450 have a synergistic effect on cerivastatin-gemfibrozil interactions. The siRNA knockdown sandwich-cultured human hepatocytes may provide a new powerful model for evaluating DDIs.
Liao
Mingxiang
M
Drug Metabolism and Pharmacokinetics, Millennium Pharmaceuticals Inc., 35 Landsdowne St., Cambridge, MA 02139, USA.
Raczynski
Arek R
AR
Chen
Michael
M
Chuang
Bei-Ching
BC
Zhu
Qing
Q
Shipman
Rob
R
Morrison
Jodi
J
Lee
David
D
Lee
Frank W
FW
Balani
Suresh K
SK
Xia
Cindy Q
CQ
D002853:Chromatography, Liquid; D004347:Drug Interactions; D022781:Hepatocytes / Q000187:drug effects* / Q000378:metabolism; D006801:Humans; D008954:Models, Biological; D027361:Organic Anion Transporters / Q000037:antagonists & inhibitors* / Q000235:genetics; D034622:RNA Interference; D012333:RNA, Messenger / Q000235:genetics; D053719:Tandem Mass Spectrometry
D027361:Organic Anion Transporters; D012333:RNA, Messenger