Multiple Efflux Pumps Are Involved in the Transepithelial Transport of Colchicine: Combined Effect of P-Glycoprotein and Multidrug Resistance-Associated Protein 2 Leads to Decreased Intestinal Absorption Throughout the Entire Small Intestine

Dahan, Arik; Sabit, Hairat; Amidon, Gordon L.

doi:10.1124/dmd.109.028282

Cited by 94 publications

(61 citation statements)

References 42 publications

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“…In intestine, indomethacin was shown to inhibit MRP2 and to increase sulfasalazine transepithelial permeability, both in rat small intestine and in Caco-2 cell monolayers [9 ]. In agreement with these indings, Caco-2 cells coincubated with indomethacin exhibited an increased permeability to the hMRP2 substrates luvastatin [9 ] and colchicine [99].…”

Section: Post-transcriptional Regulationsupporting

confidence: 71%

Hepatic and Intestinal Multidrug Resistance-Associated Protein 2: Transcriptional and Post-transcriptional Regulation by Xenobiotics

Arana¹,

Tocchetti²,

Rigalli³

et al. 2016

Toxicology - New Aspects to This Scientific Conundrum

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We are daily exposed to a large number of pharmacological drugs, environmental pollutants, and natural toxins, which represent a potential toxic insult. The organism possesses a sophisticated system of detoxiication particularly expressed in the liver, intestine, and kidney. This system consists of intracellular biotransformation enzymes that convert the toxins into more hydrophilic derivatives followed by their elimination through membrane transporters. Multidrug resistance-associated protein 2 MRP2, ABCC2 is an important member of the ATP-binding cassete ABC superfamily of transporters localized at the apical membrane of polarized cells, such as hepatocytes, enterocytes, and renal tubular cells. MRP2 is proposed as a major actor in the elimination of endo-and xenobiotics, mainly conjugated with glucuronic acid, glutathione, and sulfate. The small intestine and the liver constitute relevant detoxiication organs expressing MRP2 and therefore preventing absorption and promoting the hepatobiliary clearance of xenobiotics. MRP2 expression and/or function can be modulated by therapeutic drugs, herbal products, dietary compounds, and environmental pollutants. Consequently, MRP2 modulation could cause changes in tissue exposure, with potential toxicological and pharmacological consequences. This chapter reviews the information available on the role of hepatic and intestinal MRP2 in detoxiication processes, and their regulation by xenobiotics, considering in particular its toxicological relevance.

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Section: Post-transcriptional Regulationsupporting

confidence: 71%

Hepatic and Intestinal Multidrug Resistance-Associated Protein 2: Transcriptional and Post-transcriptional Regulation by Xenobiotics

Arana¹,

Tocchetti²,

Rigalli³

et al. 2016

Toxicology - New Aspects to This Scientific Conundrum

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show abstract

“…Similarly to fexofenadine, the efflux ratios in cell lines lacking P-gp were below unity, but were slightly increased by addition of MK571, suggesting that a basolateral MRP may interact with colchicine in the absence of P-gp. Colchicine has been reported as a substrate for both P-gp and MRP2 in Caco-2 cells and rodent intestine (Dahan et al, 2009); however, our data do not support colchicine interaction with MRP2. Reasons for this discrepancy in results may include the lack of MK571 specificity within the MRP family as well as a negative impact on the Caco-2 cell monolayer at higher concentrations, and point to the challenges in using chemical inhibitors versus gene KO technology.…”

Section: Discussioncontrasting

confidence: 57%

Zinc Finger Nuclease–Mediated Gene Knockout Results in Loss of Transport Activity for P-Glycoprotein, BCRP, and MRP2 in Caco-2 Cells

Sampson¹,

Brinker²,

Pratt³

et al. 2014

Drug Metab Dispos

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Membrane transporters P-glycoprotein [P-gp; multidrug resistance 1 (MDR1)], multidrug resistance-associated protein (MRP) 2, and breast cancer resistance protein (BCRP) affect drug absorption and disposition and can also mediate drug-drug interactions leading to safety/toxicity concerns in the clinic. Challenges arise with interpreting cell-based transporter assays when substrates or inhibitors affect more than one actively expressed transporter and when endogenous or residual transporter activity remains following overexpression or knockdown of a given transporter. The objective of this study was to selectively knock out three drug efflux transporter genes (MDR1, MRP2, and BCRP), both individually as well as in combination, in a subclone of Caco-2 cells (C2BBe1) using zinc finger nuclease technology. The wild-type parent and knockout cell lines were tested for transporter function in Transwell bidirectional assays using probe substrates at 5 or 10 mM for 2 hours at 37°C. P-gp substrates digoxin and erythromycin, BCRP substrates estrone 3-sulfate and nitrofurantoin, and MRP2 substrate 5-(and-6)-carboxy-29,79-dichlorofluorescein each showed a loss of asymmetric transport in the MDR1, BCRP, and MRP2 knockout cell lines, respectively. Furthermore, transporter interactions were deduced for cimetidine, ranitidine, fexofenadine, and colchicine. Compared with the knockout cell lines, standard transporter inhibitors showed substrate-specific variation in reducing the efflux ratios of the test compounds. These data confirm the generation of a panel of stable Caco-2 cell lines with single or double knockout of human efflux transporter genes and a complete loss of specific transport activity. These cell lines may prove useful in clarifying complex drug-transporter interactions without some of the limitations of current chemical or genetic knockdown approaches.

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“…In addition, information concerning the expression profiles of efflux transporters and metabolic enzymes along the small intestinal tract is beneficial for the effective development of orally administrated drugs. Indeed, many investigations into the localization of drug transporters and metabolic enzymes have been conducted with this goal in mind (7,8).…”

Section: Discussionmentioning

confidence: 99%

Ezrin regulates the expression of Mrp2/Abcc2 and Mdr1/Abcb1 along the rat small intestinal tract

Nakano

Sekine

Ito

et al. 2013

American Journal of Physiology-Gastrointestinal and Liver Physi

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Multidrug resistance-associated protein 2 (MRP2)/ATP-binding cassette protein C2 (ABCC2) and multidrug resistance protein 1 (MDR1)/ABCB1 are well-known efflux transporters located on the brush border membrane of the small intestinal epithelia, where they limit the absorption of a broad range of substrates. The expression patterns of MRP2/ABCC2 and MDR1/ABCB1 along the small intestinal tract are tightly regulated. Several reports have demonstrated the participation of ERM (ezrin/radixin/moesin) proteins in the posttranslational modulation of MRP2/ABCC2 and MDR1/ABCB1, especially with regard to their membrane localization. The present study focused on the in vivo expression profiles of MRP2/ABCC2, MDR1/ABCB1, ezrin, and phosphorylated ezrin to further elucidate the relationship between the efflux transporters and the ERM proteins. The current results showed good correlation between the phosphorylation status of ezrin and Mrp2/Abcc2 expression along the gastrointestinal tract of rats and between the expression profiles of both ezrin and Mdr1/Abcb1 in the small intestine. We also demonstrated the involvement of conventional protein kinase C isoforms in the regulation of ezrin phosphorylation. Furthermore, experiments conducted with wild-type (WT) ezrin and a T567A (Ala substituted Thr) dephosphorylated mutant showed a decrease in membrane surface-localized and total expressed MRP2/ABCC2 in T567A-expressing vs. WT ezrin-expressing Caco-2 cells. In contrast, T567A-and WT-expressing cells both showed an increase in membrane surface-localized and total expressed MDR1/ ABCB1. These findings suggest that the phosphorylation status and the expression profile of ezrin differentially direct MRP2/ABCC2 and MDR1/ABCB1 expression, respectively, along the small intestinal tract.ezrin; MDR1/ABCB1; MRP2/ABCC2; small intestinal tract THE SMALL INTESTINE IS A well-differentiated organ that selectively absorbs substances needed for the survival of the organism, while also functioning as a barrier to prevent the free passage of various xenobiotics (i.e., toxins, carcinogens, and drugs). Specific efflux transporters are found within the small intestinal barrier system (11), including members of the ABC protein (ATP-binding cassette) superfamily [e.g., multidrug resistance protein 1 (MDR1)/ABCB1, multidrug resistanceassociated protein 2 (MRP2)/ABCC2, and breast cancer resistance protein/ABCG2]. These efflux transporters are located on the apical membrane of the intestinal epithelium, where they convey protection to individual cells against xenobiotics and prevent the absorption of xenobiotics into the blood (6, 17).MRP2/ABCC2 and MDR1/ABCB1 are characterized by specific expression patterns in the small intestinal epithelium (6). In the rat intestine, Mrp2/Abcc2 is expressed along a gradient, with the highest levels of protein in the proximal intestine (duodenum and jejunum) and the lowest levels in the terminal ileum and colon. On the other hand, Mdr1/Abcb1 expression is lowest in the proximal region and increases in the distal directio...

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Multiple Efflux Pumps Are Involved in the Transepithelial Transport of Colchicine: Combined Effect of P-Glycoprotein and Multidrug Resistance-Associated Protein 2 Leads to Decreased Intestinal Absorption Throughout the Entire Small Intestine

Cited by 94 publications

References 42 publications

Hepatic and Intestinal Multidrug Resistance-Associated Protein 2: Transcriptional and Post-transcriptional Regulation by Xenobiotics

Hepatic and Intestinal Multidrug Resistance-Associated Protein 2: Transcriptional and Post-transcriptional Regulation by Xenobiotics

Zinc Finger Nuclease–Mediated Gene Knockout Results in Loss of Transport Activity for P-Glycoprotein, BCRP, and MRP2 in Caco-2 Cells

Ezrin regulates the expression of Mrp2/Abcc2 and Mdr1/Abcb1 along the rat small intestinal tract

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