Characterization of Transport Protein Expression in Multidrug Resistance-Associated Protein (Mrp) 2-Deficient Rats

Johnson, Brendan M.; Zhang, Peijin; Schuetz, John D.; Brouwer, Kim L. R.

doi:10.1124/dmd.105.005793

Cited by 106 publications

(96 citation statements)

References 44 publications

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“…For example, the total P450 content and activity of CYP1A1/2 and CYP2B1/2 were significantly higher in TR − compared to wild-type rats, although testosterone 6β-hydroxylation mediated by CYP3A1/2 was lower in TR − compared to wild-type rats (Newton et al, 2005;Silva et al, 2005). TR − rats exhibit up-regulated UGT1a (Johnson et al, 2006). Since Mrp2 plays an important role in GSH biliary excretion, impaired function of Mrp2 in TR − rats has been attributed to higher hepatocellular concentrations of GSH relative to wild-type rats (Lu et al, 1996).…”

Section: Discussionmentioning

confidence: 99%

“…This observation could be explained by modulation of the expression/activity of hepatic basolateral efflux transport proteins and metabolic enzymes in TR − rats. Mrp2-deficient rats are well-known to exhibit higher Mrp3 expression Ogawa et al, 2000;Johnson et al, 2006). Mrp3 mediates hepatic basolateral efflux of substrates from hepatocytes to the blood, and is thought to compensate for impaired biliary excretion of organic anions during Mrp2 deficiency Xiong et al, 2000).…”

Section: Discussionmentioning

confidence: 99%

“…Patients with Dubin-Johnson syndrome lack MRP2 and hence suffer from hereditary conjugated hyperbilirubinemia (Paulusma et al, 1997). In the absence of Mrp2/ MRP2, rats and humans exhibit increased expression of basolateral Mrp3/MRP3 as a compensatory mechanism that enables the excretion of Mrp2/MRP2 substrates into the systemic circulation, thus avoiding excessive accumulation of organic anions in the hepatocyte (Kiuchi et al, 1998;Hirohashi et al, 1999;Johnson et al, 2006;Nies and Keppler, 2007).…”

Section: Introductionmentioning

confidence: 99%

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Modulation of trabectedin (ET-743) hepatobiliary disposition by multidrug resistance-associated proteins (Mrps) may prevent hepatotoxicity

Lee

Leslie

Zamek‐Gliszczynski

et al. 2008

Toxicology and Applied Pharmacology

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Trabectedin is a promising anticancer agent, but dose-limiting hepatotoxicity was observed during phase I/II clinical trials. Dexamethasone (DEX) has been shown to significantly reduce trabectedinmediated hepatotoxicity. The current study was designed to assess the capability of sandwichcultured primary rat hepatocytes (SCRH) to predict the hepato-protective effect of DEX against trabectedin-mediated cytotoxicity. The role of multidrug resistance-associated protein 2 (Mrp2; Abcc2) in trabectedin hepatic disposition also was examined. In SCRH from wild-type Wistar rats, cytotoxicity was observed after 24-hr continuous exposure to trabectedin. SCRH pretreated with additional DEX (1 µM) exhibited a 2-3-fold decrease in toxicity at 100 nM and 1000 nM trabectedin. Unexpectedly, toxicity in SCRH from Mrp2-deficient (TR − ) compared to wild-type Wistar rats was markedly reduced. Depletion of glutathione from SCRH using buthionine sulfoximine (BSO) mitigated trabectedin toxicity associated with 100 nM and 1000 nM trabectedin. Western blot analysis demonstrated increased levels of CYP3A1/2 and Mrp2 in SCRH pretreated with DEX; interestingly, Mrp4 expression was increased in SCRH after BSO exposure. Trabectedin biliary recovery in isolated perfused livers from TR − rats was decreased by ~75% compared to wild-type livers. In conclusion, SCRH represent a useful in vitro model to predict the hepatotoxicity of trabectedin observed in vivo. The protection by DEX against trabectedin-mediated cytotoxicity may be attributed, in part, to enhanced Mrp2 biliary excretion and increased metabolism by CYP3A1/2. Decreased trabectedin toxicity in SCRH from TR − rats, and in SCRH pretreated with BSO, may be due to increased basolateral excretion of trabectedin by Mrp3 and/or Mrp4.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Modulation of trabectedin (ET-743) hepatobiliary disposition by multidrug resistance-associated proteins (Mrps) may prevent hepatotoxicity

Lee

Leslie

Zamek‐Gliszczynski

et al. 2008

Toxicology and Applied Pharmacology

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“…CL Uptake tended to decrease and CL BL tended to increase in TR 2 compared with WT SCH, consistent with the hepatoprotective compensatory changes that would be expected in the setting of impaired biliary excretion. Indeed, it has been shown that NTCP expression is downregulated and Mrp3 upregulated in livers from TR 2 compared with WT rats (Johnson et al, 2006). Transporter expression and function have been reported to change over time in rat SCH.…”

Section: Hepatic Basolateral Efflux Of Rosuvastatinmentioning

confidence: 99%

Hepatic Basolateral Efflux Contributes Significantly to Rosuvastatin Disposition I: Characterization of Basolateral Versus Biliary Clearance Using a Novel Protocol in Sandwich-Cultured Hepatocytes

Pfeifer

Yang

Brouwer

2013

The Journal of Pharmacology and Experimental Therapeutics

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103

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Transporters responsible for hepatic uptake and biliary clearance (CL Bile ) of rosuvastatin (RSV) have been well characterized. However, the contribution of basolateral efflux clearance (CL BL ) to hepatic and systemic exposure of RSV is unknown. Additionally, the appropriate design of in vitro hepatocyte efflux experiments to estimate CL Bile versus CL BL remains to be established. A novel uptake and efflux protocol was developed in sandwich-cultured hepatocytes (SCH) to achieve desired tight junction modulation while maintaining cell viability. Subsequently, studies were conducted to determine the role of CL BL in the hepatic disposition of RSV using SCH from wild-type (WT) and multidrug resistance-associated protein 2 (Mrp2)-deficient (TR 2 ) rats in the absence and presence of the P-glycoprotein and breast cancer resistance protein (Bcrp) inhibitor elacridar (GF120918). RSV CL Bile was nearly ablated by GF120918 in TR 2 SCH, confirming that Mrp2 and Bcrp are responsible for the majority of RSV CL Bile . Pharmacokinetic modeling revealed that CL BL and CL Bile represent alternative elimination routes with quantitatively similar contributions to the overall hepatocellular excretion of RSV in rat SCH under baseline conditions (WT SCH in the absence of GF120918) and also in human SCH. Membrane vesicle experiments revealed that RSV is a substrate of MRP4 (K m 5 21 6 7 mM, V max 5 1140 6 210 pmol/min per milligram of protein). Alterations in MRP4-mediated RSV CL BL due to drug-drug interactions, genetic polymorphisms, or disease states may lead to changes in hepatic and systemic exposure of RSV, with implications for the safety and efficacy of this commonly used medication.

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“…Here, we report studies that led us to the ABC subfamily C member 2 (Abcc2)-which mediates the secretion of amphiphilic glutathione, glucuronide, and sulfate conjugates into bile (5,10)-as the major relevant bile canalicular transporter of 99m Tc-mebrofenin. We performed these studies in well-defined rat models, syngeneic animals deficient in dipeptidyl peptidase IV (DPPIV) enzyme activity to serve as recipients for localizing healthy DPPIV-positive transplanted cells (9), animals with CCl 4 -induced liver injury (8), and animals mutant in the Abcc2 gene (11), which is also known as multidrug resistance protein-2.…”

mentioning

confidence: 99%

Adenosine Triphosphate–Binding Cassette Subfamily C Member 2 Is the Major Transporter of the Hepatobiliary Imaging Agent ^99mTc-Mebrofenin

Bhargava¹,

Joseph²,

Ananthanarayanan³

et al. 2009

J Nucl Med

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The organic anion 99m Tc-N-[2- [(3-bromo-2,4,6-trimethylphenyl)amino]-2-oxoethyl]-N-(carboxymethyl)-glycine ( 99m Tc-mebrofenin) and its analogs are widely used for hepatobiliary imaging. Identification of the mechanisms directing bile canalicular transport of these agents will provide insights into the basis of their hepatic handling for assessing perturbations. Methods: We performed studies in animals, including healthy Fischer 344 rats or rats treated with carbon tetrachloride or intrasplenic cell transplantation and healthy Wistar rats or HsdAMC:TR-Abcc2 mutant rats in Wistar background. Onset of hepatic inflammation was verified by analysis of carbon uptake in Kupffer cells. Hepatic clearance of 99m Tc-mebrofenin was studied with dynamic imaging, and fractional retention of peak hepatic mebrofenin activity after 60 min was determined. Changes in the expression of bile canalicular transporters were analyzed by real-time polymerase chain reaction and Western blots. Results: Carbon tetrachloride and cell transplantation produced hepatic inflammation with activation of Kupffer cells, resulting in a rapid decline in the expression of the bile canalicular transporters Abcb4, Abcb11, and Abcc2. Among these transporters, decreased expression of Abcc2 was most prominent, and this decline persisted for 4 wk. Next, we examined 99m Tc-mebrofenin excretion in HsdAMC:TR-Abcc2 mutant rats (in which Abcc2 expression is naturally inactivated), compared with their healthy counterparts. In healthy HsdRccHan:WIST rats, only 23% 6 3% of the peak 99m Tc-mebrofenin activity was retained after 60 min. By contrast, in HsdAMC:TR-Abcc2 mutant rats, 73% 6 5% of the peak 99m Tc-mebrofenin activity was retained (P , 0.001). Moreover, the administration of cyclosporin A markedly inhibited 99m Tc-mebrofenin excretion in healthy rats, with no further effect on already impaired 99m Tc-mebrofenin excretion in HsdAMC:TR-Abcc2 mutant rats. Hepatic excretion of 99m Tc-mebrofenin was largely dependent on Abcc2. This molecular basis of 99m Tc-mebrofenin excretion will advance studies of pathophysiologic mechanisms in hepatic Abcc2 pathways.

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Characterization of Transport Protein Expression in Multidrug Resistance-Associated Protein (Mrp) 2-Deficient Rats

Abstract: ABSTRACT:Multidrug resistance-associated protein (Mrp) 2-deficient transport-deficient (TR ؊ ) rats, together with their transport-competent

Cited by 106 publications

References 44 publications

Modulation of trabectedin (ET-743) hepatobiliary disposition by multidrug resistance-associated proteins (Mrps) may prevent hepatotoxicity

Modulation of trabectedin (ET-743) hepatobiliary disposition by multidrug resistance-associated proteins (Mrps) may prevent hepatotoxicity

Hepatic Basolateral Efflux Contributes Significantly to Rosuvastatin Disposition I: Characterization of Basolateral Versus Biliary Clearance Using a Novel Protocol in Sandwich-Cultured Hepatocytes

Adenosine Triphosphate–Binding Cassette Subfamily C Member 2 Is the Major Transporter of the Hepatobiliary Imaging Agent ^99mTc-Mebrofenin

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Characterization of Transport Protein Expression in Multidrug Resistance-Associated Protein (Mrp) 2-Deficient Rats

Abstract: ABSTRACT:Multidrug resistance-associated protein (Mrp) 2-deficient transport-deficient (TR ؊ ) rats, together with their transport-competent

Cited by 106 publications

References 44 publications

Modulation of trabectedin (ET-743) hepatobiliary disposition by multidrug resistance-associated proteins (Mrps) may prevent hepatotoxicity

Modulation of trabectedin (ET-743) hepatobiliary disposition by multidrug resistance-associated proteins (Mrps) may prevent hepatotoxicity

Hepatic Basolateral Efflux Contributes Significantly to Rosuvastatin Disposition I: Characterization of Basolateral Versus Biliary Clearance Using a Novel Protocol in Sandwich-Cultured Hepatocytes

Adenosine Triphosphate–Binding Cassette Subfamily C Member 2 Is the Major Transporter of the Hepatobiliary Imaging Agent 99mTc-Mebrofenin

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Product

Resources

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Adenosine Triphosphate–Binding Cassette Subfamily C Member 2 Is the Major Transporter of the Hepatobiliary Imaging Agent ^99mTc-Mebrofenin