ANTICANCER RESEARCH 34: 547-550 (2014)

Short Review Membrane Transporters as Mediators of Cisplatin Side-effects

GIULIANO CIARIMBOLI

Experimental Nephrology, Department of Internal Medicine D, and Interdisciplinary Center for Clinical Research (IZKF), University Hospital Münster, Münster, Germany

Abstract. The clinical use of the efficient chemotherapeutic Introduction. Intracellular-acting drugs must cross biological drug cisplatin is limited by its specific severe organ toxicities membranes to reach their targets. In the case of lipophilic such as nephro-, oto-, and also peripheral neurotoxicity. drugs, the passage through plasma membranes happens Membrane transporters such as the copper transporter-1 largely via passive diffusion at a rate related to their (Ctr1), the copper transporter-2 (Ctr2), the P-type copper- lipophilicity, while in the case of hydrophilic drugs this transporting ATPases ATP7A and ATP7B, the organic cation process is mediated by membrane transporters. Many transporter-2 (OCT2), and the multidrug extrusion membrane transporters have a specific tissue and cell transporter-1 (MATE1) mediate cellular transport of distribution: in the morphologically- and functionally- cisplatin. Since OCT2 is specifically expressed in the kidneys, polarized epithelial tissues, for example, transporters are even its role as possible target of specific organ protection against specifically expressed on the apical or basolateral cell undesired cisplatin toxicity is under investigation. We could membrane. In this way, a specific drug-transporter interaction show that OCT2 is also expressed in the cochlea in hair cells can be exploited to target drugs to selected cells and tissues, and in cells of the stria vascularis and also in dorsal root but of course can also explain specific undesired adverse ganglia of mice. Moreover, we could show in a mouse model effects (1). While the pharmacological relevance of transport of cisplatin acute toxicities that the expression of OCT is proteins is well-known because of their role in the critical for the development of ototoxicity, peripheral development of resistance of tumor cells to chemotherapeutic neurotoxicity and nephrotoxicity. Competition of cisplatin agents (as for example in the case of P-glycoprotein (2)), their transport by the OCT2 substrate cimetidine was able to role in the development of specific drug adverse effects has suppress ototoxicity, and reduce nephrotoxicity. Only few been only in the recent years under critical investigation (1). human tumors express OCT2, its expression being apparently Cisplatin is a potent and efficient cytostatic drug, whose down-regulated by epigenetic modifications, suggesting that clinical use is limited by its severe acute and chronic nephro-, a protective therapy by competition for the transport of oto- and peripheral neurotoxicity (3). In this work I cisplatin by OCT2 may be generally feasible without summarize our results presented at the the XIth International affecting its antitumor potency. There is already some Symposium on Platin Coordination Compound in Cancer evidence that patients bearing a mutation in OCT2 gene or held in Verona (Italy) on the role of membrane transporters co-medicated with cimetidine are protected against cisplatin in mediating specific cisplatin uptake and toxicities and the nephrotoxicity. In conclusion, OCT2 seems to be an ideal thereby possible implications for protective therapies. target for the establishment of protective therapies aimed to specifically reduce cisplatin side-effects and increase the Cisplatin toxicities. Cisplatin treatment, even though quality of life of the patients. effective against tumors, has severe side-effects such as nephrotoxicity, which is often dose-limiting, ototoxicity, and peripheral neurotoxicity. Clinical signs of acute and/or chronic nephrotoxicity are a decrease in renal plasma flow Correspondence to: Giuliano Ciarimboli, Ph.D., Medizinische Klinik (4) and in glomerular filtration rate (5), an increase in serum D, Experimentelle Nephrologie, Universitätsklinikum Münster, creatinine, and reduction of serum magnesium and Albert-Schweitzer-Campus 1, Gebäude A14, 48149 Münster, potassium levels (6). Even though nephrotoxicity can be Germany. Tel: +49 2518356981, Fax: +49 2518356973, e-mail: [email protected] controlled by diuretics and pre-hydration of patients, the prevalence of cisplatin nephrotoxicity is still high, occurring Key Words: Transporters, cisplatin, toxicities, review, membrane in about one-third of patients undergoing cisplatin treatment transporters, Ctr2, OCT2, MATE1. (6). Studies in experimental animals showed that the kidney

0250-7005/2014 $2.00+.40 547 ANTICANCER RESEARCH 34: 547-550 (2014) accumulates more cisplatin than other organs (7), and that (16). Moreover, competition of hOCT2 transport by the OCT the proximal tubules are the renal structure specifically substrate cimetidine effectively suppressed cisplatin cellular damaged by cisplatin (8). Ototoxicity, an untypical side- toxicity in vitro (16). The introduction of mice, where the effect for a chemotherapeutic drug, is caused by a damage genes for OCT1 and OCT2 were genetically deleted (OCT1 first occurring at the cochlea base, where high-frequency and 2 knock-out -ko- mice) (25) allowed us (17) and others sounds are processed, which, with increasing cumulative (26) to study the importance of these transporters for the dose, proceeds towards the apex, affecting also hearing at development of acute cisplatin toxicities in vivo. Compared lower frequencies (9). Cisplatin treatment causes hearing with wild-type mice, OCT1 and 2 ko mice developed a much loss, possibly leading to deafness, which is still an milder nephrotoxicity under acute treatment with cisplatin unresolved clinical problem of dramatic importance in (17). Interestingly, these mice were also protected against infants and younger children, where ototoxicity can lead to cisplatin induced ototoxicity (17). Because of this finding, a delayed language development which in turn might have we investigated the expression of OCT2 in the mouse devastating consequences for a young child’s social and cochlea and were able to show its expression in hair cells educational development (10). Most patients treated with and in the cells of the stria vascularis, structures sensitive to cisplatin develop a symptomatic and clinically-detectable cisplatin damage. Recently, performing laser-ablation mass sensory neuropathy, caused by its preferential uptake in the spectrometry experiments in wildtype mice acutely treated dorsal root ganglia (DRG), which produces a dose-related with cisplatin, we were able to show a high accumulation of large fiber sensory neuropathy (11, 12). cisplatin in the kidney cortex and also in the stria vascularis of the cochlea, areas where OCT are specifically expressed Cisplatin transporters. Several different transporters seem (27). Interestingly, treatment of the wildtype animals with to be involved in the cellular transport of cisplatin: the cisplatin together with cimetidine, a competitor for the copper transporter-1 (Ctr1) (13), the copper transporter-2 transport by OCT, eliminated or lowered the ototoxic and (Ctr2) (14), the P-type copper-transporting ATPases nephrotoxic effects of cisplatin, respectively (17). The reason ATP7A and ATP7B (15), the organic cation transporter-2 for incomplete protection from cisplatin nephrotoxicity by (OCT2) (16, 17), and the multidrug and toxin extrusion cimetidine may reside in the vectorial nature of cisplatin transporter 1 (MATE1) (18, 19). While Ctr1, Ctr2, ATP7A transport in renal proximal tubules: cimetidine inhibits with and ATP7B are ubiquitary expressed, OCTs and MATE1 higher affinity the apical expressed MATE1 (28), probably are highly expressed in secretory organs, such as the liver an efflux transporter for cisplatin, in this way probably and the kidneys, and for this reason, are of special interest reducing its renoprotective effect. A study in rats showed that as mediators of specific organ toxicities. Focussing on such a protective strategy did not interfere with the anti- OCTs, these transporters have a species- and subtype- tumoral activity of cisplatin (29). Interestingly, OCT2 has specific organ expression: human OCT2 (hOCT2) is been demonstrated to be expressed in DRG within the specifically expressed in the basolateral membrane of renal nervous system (30). Genetic or pharmacological knockout proximal tubule cells, while human hepatocytes express of OCT2 protected mice from hypersensitivity to cold or mainly hOCT1 in their sinusoidal membrane (20). For the mechanical-induced allodynia induced by oxaliplatin right interpretation of translational studies, it is important treatment (30). Even though the idea of a specific protective to mention that in rodents, both OCT1 and OCT2 show a therapy aimed to reduce cisplatin toxicities by competing its high renal expression in the basolateral membrane of uptake by OCT is very attractive, it is important not to proximal tubule cells, with higher OCT2 expression in compromise its uptake in tumor cells, the target of cisplatin male animals (21,22). therapy. Investigating the transporter expression in several We furnished the first direct in vitro evidences for the different tumor derived cell lines, we found a strong transport of cisplatin by hOCT2 showing that it inhibits the expression of Ctr1 but no or only a very low expression of uptake of the fluorescent organic cation 4-(4-(dimethylamino) OCT, at least at the mRNA level (17). However, colorectal styryl)-N-methylpyridinium (ASP+) by hOCT2 stably tumor samples expressed hOCT2 (31), while a reduced transfected in human embryonic kidney (HEK) cells. The expression of OCT1, OCT2, and OCT3 due to epigenetic expression of hOCT2 was also linked to a higher cisplatin modifications such as DNA methylation was observed both accumulation compared to wild-type HEK cells (16). These in animal models and in prostate and liver tumor patients findings were confirmed by other groups studying cisplatin (32-34). There are few studies in patients suggesting a uptake by rat OCT2 (23) and by hOCT2 (24). We could also protective effect of cimetidine against and an important role showed that cisplatin competed with ASP+ basolateral for hOCT2 for the development of cisplatin nephrotoxicity. transport in freshly-isolated human proximal tubules (where One study suggested already 1987 an efficient hOCT2 is expressed), whereas it did not influence ASP+ nephroprotection by cimetidine in patients treated with transport in human hepatocytes (where hOCT1 is expressed) cisplatin, without compromising the success of tumor

548 Ciarimboli: Membrane Transporters as Mediators of Cisplatin Side-effects (Short Review) therapy (35). Another study showed that patients bearing a 14 Blair BG, Larson CA, Safaei R and Howell SB: Copper transporter mutation in hOCT2, which decreases transport function, were 2 regulates the cellular accumulation and cytotoxicity of Cisplatin protected against cisplatin nephrotoxicity (26). A recent and Carboplatin. Clin Cancer Res 15: 4312-4321, 2009. study showed that both co-medication with cimetidine and 15 Samimi G, Katano K, Holzer AK, Safaei R and Howell SB: Modulation of the cellular pharmacology of cisplatin and its genetic polymorphism of hOCT2 protected patients against analogs by the copper exporters ATP7A and ATP7B. Mol cisplatin nephrotoxicity (36). Pharmacol 66: 25-32, 2004. Collectively, these results indicate the critical importance 16 Ciarimboli G, Ludwig T, Lang D, Pavenstädt H, Koepsell H, of OCT2 in the handling of cisplatin in the kidney, in the Piechota HJ, Haier J, Jaehde U, Zisowsky J and Schlatter E: inner ear and possibly also in DRG and provide a rationale Cisplatin nephrotoxicity is critically mediated via the human for the development of new targeted approaches to mitigate organic cation transporter 2. Am J Pathol 167: 1477-1484, 2005. these debilitating side effects of cisplatin chemotherapy. 17 Ciarimboli G, Deuster D, Knief A, Sperling M, Holtkamp M, Edemir B, Pavenstädt H, Lanvers-Kaminsky C, Am Zehnhoff- Dinnesen A, Schinkel AH, Koepsell H, Jurgens H and Schlatter Acknowledgements E: Organic cation transporter 2 mediates cisplatin-induced oto- and nephrotoxicity and is a target for protective interventions. The work of the Author on this topic is supported by the Am J Pathol 176: 1169-1180, 2010. Interdisciplinary Center for Clinical Research (IZKF) Münster 18 Yonezawa A, Masuda S, Yokoo S, Katsura T and Inui K: (Grant Cia2/013/13) and by the Deutsche Krebshilfe (grant 108539). Cisplatin and oxaliplatin, but not carboplatin and nedaplatin, are substrates for human organic cation transporters (SLC22A1-3 References and multidrug and toxin extrusion family). J Pharmacol Exp Ther 319: 879-886, 2006. 1 Ciarimboli G: Role of organic cation transporters in drug- 19 Nakamura T, Yonezawa A, Hashimoto S, Katsura T and Inui KI: induced toxicity. Expert Opin Drug Metab Toxicol 7: 159-174, Disruption of multidrug and toxin extrusion MATE1 potentiates 2011. cisplatin-induced nephrotoxicity. Biochem Pharmacol 80: 1762- 2 Zhou SF: Structure, function and regulation of P-glycoprotein 1767, 2010. and its clinical relevance in drug disposition. Xenobiotica 38: 20 Ciarimboli G: Organic cation transporters. Xenobiotica 38: 936- 802-832, 2008. 971, 2008. 3 Rabik CA and Dolan ME: Molecular mechanisms of resistance 21 Urakami Y, Nakamura N, Takahashi K, Okuda M, Saito H, and toxicity associated with platinating agents. Cancer Treat Rev Hashimoto Y and Inui K: Gender differences in expression of 33: 9-23, 2007. organic cation transporter OCT2 in rat kidney. FEBS Lett 461: 4 Fillastre JP and Raguenez-Viotte G: Cisplatin nephrotoxicity. 339-342, 1999. Toxicol Lett 46: 163-175, 1989. 22 Meetam P, Srimaroeng C, Soodvilai S and Chatsudthipong V: 5 Daugaard G: Cisplatin nephrotoxicity: experimental and clinical Regulatory role of testosterone in organic cation transport: in studies. Dan Med Bull 37: 1-12, 1990. vivo and in vitro studies. Biol Pharm Bull 32: 982-987, 2009. 6 Pabla N and Dong Z: Cisplatin nephrotoxicity: mechanisms and 23 Yonezawa A, Masuda S, Nishihara K, Yano I, Katsura T and Inui renoprotective strategies. Kidney Int 73: 994-1007, 2008. K: Association between tubular toxicity of cisplatin and 7 Litterst CL, Gram TE, Dedrick RL, LeRoy AF and Guarino AM: expression of organic cation transporter rOCT2 (Slc22a2) in the Distribution and disposition of platinum following intravenous rat. Biochem Pharmacol 70: 1823-1831, 2005. administration of cis-diamminedichloroplatinum(II) (NSC 24 Filipski KK, Loos WJ, Verweij J and Sparreboom A: Interaction 119875) to dogs. Cancer Res 36: 2340-2344, 1976. of Cisplatin with the human organic cation transporter 2. Clin 8 Dobyan DC, Levi J, Jacobs C, Kosek J and Weiner MW: Cancer Res 14: 3875-3880, 2008. Mechanism of cis-platinum nephrotoxicity: II. Morphologic 25 Jonker JW, Wagenaar E, Van Eijl S and Schinkel AH: Deficiency observations. J Pharmacol Exp Ther 213: 551-556, 1980. in the organic cation transporters 1 and 2 (Oct1/Oct2 9 Rybak LP: Mechanisms of cisplatin ototoxicity and progress in [Slc22a1/Slc22a2]) in mice abolishes renal secretion of organic otoprotection. Curr Opin Otolaryngol Head Neck Surg 15: 364- cations. Mol Cell Biol 23: 7902-7908, 2003. 369, 2007. 26 Filipski KK, Mathijssen RH, Mikkelsen TS, Schinkel AH and 10 Skinner R: Best practice in assessing ototoxicity in children with Sparreboom A: Contribution of organic cation transporter 2 cancer. Europ J Cancer 40: 2352-2354, 2004. (OCT2) to cisplatin-induced nephrotoxicity. Clin Pharmacol 11 Albers JW, Chaudhry V, Cavaletti G, and Donehower RC: Ther 86: 396-402, 2009. Interventions for preventing neuropathy caused by cisplatin and 27 Reifschneider O, Wehe CA, Raj I, Ehmcke J, Ciarimboli G, related compounds. Cochrane Database Syst Rev CD005228, 2011. Sperling M and Karst U: Quantitative bioimaging of platinum in 12 Meijer C, de Vries EG, Marmiroli P, Tredici G, Frattola L and polymer embedded mouse organs using laser ablation ICP-MS. Cavaletti G: Cisplatin-induced DNA-platination in experimental Metallomics 5: 1440-1447, 2013. dorsal root ganglia neuronopathy. Neurotoxicology 20: 883-887, 28 Ito S, Kusuhara H, Yokochi M, Toyoshima J, Inoue K, Yuasa H, 1999. and Sugiyama Y: Competitive inhibition of the luminal efflux by 13 Ishida S, Lee J, Thiele DJ and Herskowitz I: Uptake of the multidrug and toxin extrusions, but not basolateral uptake by anticancer drug cisplatin mediated by the copper transporter Ctr1 organic cation transporter 2, is the likely mechanism underlying in yeast and mammals. Proc Natl Acad Sci USA 99: 14298- the pharmacokinetic drug-drug interactions caused by cimetidine 14302, 2002. in the kidney. J Pharmacol Exp Ther 340: 393-403, 2012.

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29 Katsuda H, Yamashita M, Katsura H, Yu J, Waki Y, Nagata N, 34 Chen L, Hong C, Chen EC, Yee SW, Xu L, Almof EU, Wen C, Sai Y and Miyamoto K: Protecting Cisplatin-induced Fujii K, Johns SJ, Stryke D, Ferrin TE, Simko J, Chen X, nephrotoxicity with cimetidine does not affect antitumor activity. Costello JF and Giacomini KM: Genetic and epigenetic Biol Pharm Bull 33: 1867-1871, 2010. regulation of the organic cation transporter 3, SLC22A3. 30 Sprowl JA, Ciarimboli G, Lancaster CS, Giovinazzo H, Gibson Pharmacogenomics J 13: 110-120, 2013. AA, Du G, Janke LJ, Cavaletti G, Shields AF and Sparreboom 35 Sleijfer DT, Offerman JJ, Mulder NH, Verweij M, van der Hem A: Oxaliplatin-induced neurotoxicity is dependent on the organic GK, Schraffordt Koops HS and Meijer S: The protective cation transporter OCT2. Proc Natl Acad Sci USA 110: 11199- potential of the combination of verapamil and cimetidine on 11204, 2013. cisplatin-induced nephrotoxicity in man. Cancer 60: 2823-2828, 31 Zhang S, Lovejoy KS, Shima JE, Lagpacan LL, Shu Y, Lapuk 1987. A, Chen Y, Komori T, Gray JW, Chen X, Lippard SJ and 36 Zhang J and Zhou W: Ameliorative effects of SLC22A2 gene Giacomini KM: Organic cation transporters are determinants of polymorphism 808 G/T and cimetidine on cisplatin-induced oxaliplatin cytotoxicity. Cancer Res 66: 8847-8857, 2006. nephrotoxicity in Chinese cancer patients. Food Chem Toxicol 32 Yang J, Kalogerou M, Gallacher J, Sampson JR and Shen MH: 50: 2289-2293, 2012. Renal tumours in a Tsc1+/– mouse model show epigenetic suppression of organic cation transporters Slc22a1, Slc22a2 and Slc22a3, and do not respond to metformin. Eur J Cancer 49: 1479-1490, 2013. 33 Schaeffeler E, Hellerbrand C, Nies AT, Winter S, Kruck S, Hofmann U, van der Kuip H, Zanger UM, Koepsell H and Schwab M: DNA methylation is associated with downregulation Received November 6, 2013 of the organic cation transporter OCT1 (SLC22A1) in human Revised December 6, 2013 hepatocellular carcinoma. Genome Med 3: 82, 2011. Accepted December 9, 2013

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