Knocking Down Breast Cancer Resistance Protein (Bcrp) by Adenoviral Vector-Mediated RNA Interference (RNAi) in Sandwich-Cultured Rat Hepatocytes: A Novel Tool To Assess the Contribution of Bcrp to Drug Biliary Excretion

Yue, Wei; Abe, Kōji; Brouwer, Kim L. R.

doi:10.1021/mp800100e

Cited by 40 publications

(58 citation statements)

References 53 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Brain uptake of digoxin was approximately 35 times larger in P-gp knockout mice as compared to wildtype control mice, which indicated that P-gp is responsible for the low distribution of digoxin to the brain in vivo [57]. Digoxin is not a substrate for Bcrp [58], but other studies have indicated that the poor BBB permeability of digoxin is not explained by P-gp alone [11,38]. Verapamil has been used as a P-gp inhibitor in concentrations from 5 to 150 μM [23,[59][60][61], but it has also been shown to inhibit Mrp activity in concentrations of 5-10 μM [45,62] as well as Bcrp activity, although not in the concentration range applied here [63,64].…”

Section: Efflux Transporter Activity In Bovine Blood-brain Barrier Momentioning

confidence: 99%

An Electrically Tight In Vitro Blood–Brain Barrier Model Displays Net Brain-to-Blood Efflux of Substrates for the ABC Transporters, P-gp, Bcrp and Mrp-1

Helms

Hersom

Kuhlmann

et al. 2014

AAPS J

View full text Add to dashboard Cite

Abstract. Efflux transporters of the ATP-binding cassette superfamily including breast cancer resistance protein (Bcrp/Abcg2), P-glycoprotein (P-gp/Abcb1) and multidrug resistance-associated proteins (Mrp's/ Abcc's) are expressed in the blood-brain barrier (BBB). The aim of this study was to investigate if a bovine endothelial/rat astrocyte in vitro BBB co-culture model displayed polarized transport of known efflux transporter substrates. The co-culture model displayed low mannitol permeabilities of 0.95±0.1·10 H-etoposide revealed polarized transport favouring the brain-to-blood direction for all substrates. Steady state efflux ratios of 2.5±0.2 for digoxin, 4.4±0.5 for estrone-3-sulphate and 2.4±0.1 for etoposide were observed. These were reduced to 1.1±0.08, 1.4±0.2 and 1.5±0.1, by addition of verapamil (digoxin), Ko143 (estrone-3-sulphate) or zosuquidar+reversan (etoposide), respectively. Brain-toblood permeability of all substrates was investigated in the presence of the efflux transporter inhibitors verapamil, Ko143, zosuquidar, reversan and MK 571 alone or in combinations. Digoxin was mainly transported via P-gp, estrone-3-sulphate via Bcrp and Mrp's and etoposide via P-gp and Mrp's. The expression of P-gp, Bcrp and Mrp-1 was confirmed using immunocytochemistry. The findings indicate that P-gp, Bcrp and at least one isoform of Mrp are functionally expressed in our bovine/rat co-culture model and that the model is suitable for investigations of small molecule transport.KEY WORDS: blood-brain barrier; breast cancer resistance protein; multidrug resistance-associated protein; p-glycoprotein; polarized small molecule transport.

show abstract

Section: Efflux Transporter Activity In Bovine Blood-brain Barrier Momentioning

confidence: 99%

An Electrically Tight In Vitro Blood–Brain Barrier Model Displays Net Brain-to-Blood Efflux of Substrates for the ABC Transporters, P-gp, Bcrp and Mrp-1

Helms

Hersom

Kuhlmann

et al. 2014

AAPS J

View full text Add to dashboard Cite

show abstract

“…OSM exposure is however 15 likely to result in reduced sinusoidal uptake of drugs, owing to its global repressing effect toward influx SLC transporters. In addition, OSM-mediated down-regulation of ABCG2 expression may contribute to decreased biliary secretion of drugs such as the antibiotic nitrofurantoin handled by this canalicular ABC pump [34]. Associated with the known repression of drug metabolizing CYP activity triggerred by OSM, these putative alterations of hepatic drug transport may contribute to alteration of pharmacokinetics.…”

Section: Discussionmentioning

confidence: 99%

Regulation of drug transporter expression by oncostatin M in human hepatocytes

Vée

Jouan

Stieger³

et al. 2011

Biochemical Pharmacology

View full text Add to dashboard Cite

The cytokine oncostatin M (OSM) is a member of the interleukin (IL)-6 family, known to down-regulate expression of drug metabolizing cytochromes P-450 in human hepatocytes. The present study was designed to determine whether OSM may also impair expression of sinusoidal and canalicular drug transporters, which constitute important determinants of drug hepatic clearance. Exposure of primary human hepatocytes to OSM down-regulated mRNA levels of major sinusoidal solute carrier (SLC) influx transporters, including sodium-taurocholate co-transporting polypeptide (NTCP), organic anion transporting polypeptide (OATP) 1B1, OATP1B3, OATP2B1, organic cation transporter 1 and organic anion transporter 2. OSM also repressed mRNA expressions of ATP binding cassette (ABC) efflux transporters such as multidrug resistance protein (MRP) 2/ABCC2 and breast cancer resistance protein/ABCG2, without however impairing those of multidrug resistance gene 1/P-glycoprotein/ABCB1, MRP3/ABCC3, MRP4/ABCC4 and bile salt export pump/ABCB11. The cytokine concomitantly reduced NTCP, OATP1B1, OATP2B1 and ABCG2 protein expression and NTCP and OATP transport activities. OSM effects towards transporters were found to be dose-dependent and highly correlated with those of IL-6, but not with those of other inflammatory cytokines such as tumor necrosis factor-α or interferon-γ. In addition, OSM-mediated repression of some transporters such as NTCP, OATP1B1 and OATP2B1, was counteracted by knocking-down expression of the type II OSM receptor subunits through siRNA transfection. This OSM-mediated down-regulation of drug SLC transporters and ABCG2 in human hepatocytes may contribute to alterations of pharmacokinetics in patients suffering from diseases associated with increased production of OSM. This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain. A c c e p t e d M a n u s c r i p t 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 61 62 63 64 65 2 Accepted Manuscript ABSTRACTThe cytokine oncostatin M (OSM) is a member of the interleukin (IL)-6 family, known to down-regulate expression of drug metabolizing cytochromes P-450 in human hepatocytes.The present study was designed to determine whether OSM may also impair expression of sinusoidal and canalicular drug transporters, which constitute important determinants of drug

show abstract

“…Short interfering RNA (siRNA) can mediate strong and specific suppression of gene expression by sequencespecific cleavage of mRNA, thus blocking the translation into target protein (Yue et al, 2009). The gene silencing technique has great potential in drug efflux and transport-mediated drug-drug interactions studies for elucidating the function of specific transporters in drug disposition.…”

Section: Discussionmentioning

confidence: 99%

“…The use of the post-transcriptional gene silencing by RNA interference to specifically knock down drug transporters is a rapidly growing field of research (Celius et al, 2004;Hua et al, 2005;Tian et al, 2005;Watanabe et al, 2005;Li et al, 2006;Yue et al, 2009;Zhang et al, 2009). Even though siRNA and shRNA have emerged as powerful tools to study gene function in a sequence-specific manner, off-target effects have been observed in several studies (Jackson et al,FIG.…”

Section: Discussionmentioning

confidence: 99%

Investigation of the Involvement of P-Glycoprotein and Multidrug Resistance-Associated Protein 2 in the Efflux of Ximelagatran and Its Metabolites by Using Short Hairpin RNA Knockdown in Caco-2 Cells

Darnell

Karlsson²,

Owen³

et al. 2009

Drug Metab Dispos

View full text Add to dashboard Cite

ABSTRACT:Liver and bile secretion can be an important first-pass and clearance route for drug compounds and also the site of several drug-drug interactions. In the clinical program for ximelagatran development, an unexpected effect of erythromycin on the pharmacokinetics of the direct thrombin inhibitor ximelagatran and its metabolites was detected. This interaction was believed to be mediated by inhibition of drug transporters, which normally extrude the drug into the bile. Previous Caco-2 cell experiments indicated the involvement of an active efflux mechanism for ximelagatran, hydroxy-melagatran, and melagatran possibly mediated by P-glycoprotein (P-gp). However, the inhibitors used may not have been specific enough and the possibility that transporters other than P-gp were important in the Caco-2 cell assay cannot be excluded. In this study we used RNA interference, a post-transcriptional gene silencing mechanism in which mRNA is degraded in a sequence-specific manner, to specifically knock down P-gp or multidrug resistance-associated protein 2 (MRP2) transporters in Caco-2 cells. The data obtained from bidirectional transport studies in these cells indicate a clear involvement of P-gp but not of MRP2 in the transport of ximelagatran, hydroxy-melagatran, and melagatran across the apical cell membrane. The present study shows that short hairpin RNA Caco-2 cells are a valuable tool to investigate the contribution of specific transporters in the transcellular transport of drug molecules and to predict potential sites of pharmacokinetic interactions. The results also suggest that inhibition of hepatic P-gp is involved in the erythromycin-ximelagatran interaction seen in clinical studies.The direct thrombin inhibitor ximelagatran was envisaged to have a low potential for pharmacokinetic drug-drug interactions (DDIs) based on the preclinical information showing that ximelagatran, its intermediate metabolites, and melagatran are neither substrates nor inhibitors of the major cytochrome P450 isoenzymes (Bredberg et al., 2003). The preclinical results were supported by the results from three specific healthy volunteer studies showing no clinically significant DDIs between ximelagatran and diclofenac, diazepam, or nifedipine when coadministered simultaneously (Bredberg et al., 2003). The phase I DDI study showing a clear interaction with erythromycin, therefore, came as a surprise. Melagatran area under the curve increased by 82%, whereas ximelagatran plasma levels seemed to be essentially unchanged. Ximelagatran is bioconverted to melagatran via two intermediates, hydroxy-melagatran and ethyl-melagatran (Fig. 1), and an elevation in the plasma concentrations was seen for both intermediates, after coadministration with erythromycin (Eriksson et al., 2006).Several in vivo and in vitro studies have been performed to investigate the nature of the erythromycin-ximelagatran interaction (Eriksson et al., 2006; Sjö din et al., 2008). The pharmacokinetic profile in humans indicated that the elevation of ximelagatran and its m...

show abstract

Knocking Down Breast Cancer Resistance Protein (Bcrp) by Adenoviral Vector-Mediated RNA Interference (RNAi) in Sandwich-Cultured Rat Hepatocytes: A Novel Tool To Assess the Contribution of Bcrp to Drug Biliary Excretion

Cited by 40 publications

References 53 publications

An Electrically Tight In Vitro Blood–Brain Barrier Model Displays Net Brain-to-Blood Efflux of Substrates for the ABC Transporters, P-gp, Bcrp and Mrp-1

An Electrically Tight In Vitro Blood–Brain Barrier Model Displays Net Brain-to-Blood Efflux of Substrates for the ABC Transporters, P-gp, Bcrp and Mrp-1

Regulation of drug transporter expression by oncostatin M in human hepatocytes

Investigation of the Involvement of P-Glycoprotein and Multidrug Resistance-Associated Protein 2 in the Efflux of Ximelagatran and Its Metabolites by Using Short Hairpin RNA Knockdown in Caco-2 Cells

Contact Info

Product

Resources

About