Advertisement

The AAPS Journal

, 21:88 | Cite as

Optimized Renal Transporter Quantification by Using Aquaporin 1 and Aquaporin 2 as Anatomical Markers: Application in Characterizing the Ontogeny of Renal Transporters and Its Correlation with Hepatic Transporters in Paired Human Samples

  • Cindy Yanfei Li
  • Chelsea Hosey-Cojocari
  • Abdul Basit
  • Jashvant D. Unadkat
  • J. Steven Leeder
  • Bhagwat PrasadEmail author
Research Article

Abstract

Renal transporters, which are primarily located in the proximal tubules, play an important role in secretion and nephrotoxicity of drugs. The goal of this study was to characterize the age-dependent protein abundance of human renal transporters. A total of 43 human kidneys, 26 of which were paired with livers from the same donors, were obtained and classified into three age groups: children (< 12 years), adolescents (12 to < 18 years), and adults (> 18 years). Protein abundance of kidney-specific anatomical markers, aquaporins 1 and 2 (markers of proximal and distal/collecting tubules, respectively), and 17 transporters was quantified by LC-MS/MS proteomics. Six out of 43 kidney samples were identified as outliers (Grubbs’ test) that were significantly different from the others with relatively higher aquaporin 2 to aquaporin 1 ratio, indicating that these cortex samples were likely contaminated by medulla (representing distal/collecting tubules). No significant age-related changes (age > 1 year) were observed for renal transporter abundance, albeit OCT2 abundance was modestly higher (< 50%) in adolescents than that in adults. Higher protein-protein correlation between transporters was observed in the kidney but abundance of transporters between tissues was not correlated. The use of aquaporins 1 and 2 provides a method for identifying kidney cortex with significant contamination from medulla containing distal and collecting tubules. The abundance and protein-protein correlation data can be used in physiologically based pharmacokinetic (PBPK) modeling and simulation of renal drug disposition and clearance in pediatric populations.

KEY WORDS

ontogeny renal transporters liver kidney quantitative proteomics paired samples 

Notes

Acknowledgments

We thank Matthew Karasu from University of Washington for technical assistance with LC-MS/MS sample preparation, and Wendy Wang, Chengpeng Bi, Jeffrey Johnston, and Neil Miller from Children’s Mercy Hospital for their contribution to the RNA-seq processing.

Funding information

This study was supported by UWRAPT (University of Washington Research Affiliate Program on Transporters sponsored by Biogen, Genentech, Gilead, Takeda and Merck & Co., Inc). BP was supported by Eunice Kennedy Shriver National Institute of Child Health and Human Development, NIH grant (R01.HD081299). The National Institute of Child Health and Human Development Brain and Tissue Bank for Developmental Disorders at the University of Maryland is funded by NIH contract HHSN275200900011C, reference no. N01-HD-9-0011.

Supplementary material

12248_2019_359_MOESM1_ESM.docx (482 kb)
ESM 1 (DOCX 481 kb)

References

  1. 1.
    Morrissey KM, Stocker SL, Wittwer MB, Xu L, Giacomini KM. Renal transporters in drug development. Annu Rev Pharmacol Toxicol. 2013;53:503–29.CrossRefGoogle Scholar
  2. 2.
    Yin J, Wang J. Renal drug transporters and their significance in drug-drug interactions. Acta Pharm Sin B. 2016;6(5):363–73.CrossRefGoogle Scholar
  3. 3.
    International Transporter C, Giacomini KM, Huang SM, Tweedie DJ, Benet LZ, Brouwer KL, et al. Membrane transporters in drug development. Nat Rev Drug Discov. 2010;9(3):215–36.CrossRefGoogle Scholar
  4. 4.
    Leeder JS, Spino M, Isles AF, Tesoro AM, Gold R, MacLeod SM. Ceftazidime disposition in acute and stable cystic fibrosis. Clin Pharmacol Ther. 1984;36(3):355–62.CrossRefGoogle Scholar
  5. 5.
    Filipski KK, Mathijssen RH, Mikkelsen TS, Schinkel AH, Sparreboom A. Contribution of organic cation transporter 2 (OCT2) to cisplatin-induced nephrotoxicity. Clin Pharmacol Ther. 2009;86(4):396–402.CrossRefGoogle Scholar
  6. 6.
    Miller RP, Tadagavadi RK, Ramesh G, Reeves WB. Mechanisms of cisplatin nephrotoxicity. Toxins (Basel). 2010;2(11):2490–518.CrossRefGoogle Scholar
  7. 7.
    Hawwa AF, McKiernan PJ, Shields M, Millership JS, Collier PS, McElnay JC. Influence of ABCB1 polymorphisms and haplotypes on tacrolimus nephrotoxicity and dosage requirements in children with liver transplant. Br J Clin Pharmacol. 2009;68(3):413–21.CrossRefGoogle Scholar
  8. 8.
    Prasad B, Johnson K, Billington S, Lee C, Chung GW, Brown CD, et al. Abundance of drug transporters in the human kidney cortex as quantified by quantitative targeted proteomics. Drug Metab Dispos. 2016;44(12):1920–4.CrossRefGoogle Scholar
  9. 9.
    Kumar V, Yin J, Billington S, Prasad B, Brown CDA, Wang J, et al. The importance of incorporating OCT2 plasma membrane expression and membrane potential in IVIVE of metformin renal secretory clearance. Drug Metab Dispos. 2018;46(10):1441–5.CrossRefGoogle Scholar
  10. 10.
    Brouwer KL, Aleksunes LM, Brandys B, Giacoia GP, Knipp G, Lukacova V, et al. Human ontogeny of drug transporters: review and recommendations of the Pediatric Transporter Working Group. Clin Pharmacol Ther. 2015;98(3):266–87.CrossRefGoogle Scholar
  11. 11.
    Prasad B, Gaedigk A, Vrana M, Gaedigk R, Leeder JS, Salphati L, et al. Ontogeny of hepatic drug transporters as quantified by LC-MS/MS proteomics. Clin Pharmacol Ther. 2016;100(4):362–70.CrossRefGoogle Scholar
  12. 12.
    Agarwal SK, Gupta A. Aquaporins: the renal water channels. Indian J Nephrol. 2008;18(3):95–100.CrossRefGoogle Scholar
  13. 13.
    Nielsen S, Frokiaer J, Marples D, Kwon TH, Agre P, Knepper MA. Aquaporins in the kidney: from molecules to medicine. Physiol Rev. 2002;82(1):205–44.CrossRefGoogle Scholar
  14. 14.
    Deo AK, Prasad B, Balogh L, Lai Y, Unadkat JD. Interindividual variability in hepatic expression of the multidrug resistance-associated protein 2 (MRP2/ABCC2): quantification by liquid chromatography/tandem mass spectrometry. Drug Metab Dispos. 2012;40(5):852–5.CrossRefGoogle Scholar
  15. 15.
    Prasad B, Lai Y, Lin Y, Unadkat JD. Interindividual variability in the hepatic expression of the human breast cancer resistance protein (BCRP/ABCG2): effect of age, sex, and genotype. J Pharm Sci. 2013;102(3):787–93.CrossRefGoogle Scholar
  16. 16.
    Prasad B, Evers R, Gupta A, Hop CE, Salphati L, Shukla S, et al. Interindividual variability in hepatic organic anion-transporting polypeptides and P-glycoprotein (ABCB1) protein expression: quantification by liquid chromatography tandem mass spectroscopy and influence of genotype, age, and sex. Drug Metab Dispos. 2014;42(1):78–88.CrossRefGoogle Scholar
  17. 17.
    Nomura M, Motohashi H, Sekine H, Katsura T, Inui K. Developmental expression of renal organic anion transporters in rat kidney and its effect on renal secretion of phenolsulfonphthalein. Am J Physiol Renal Physiol. 2012;302(12):F1640–9.CrossRefGoogle Scholar
  18. 18.
    Buist SC, Klaassen CD. Rat and mouse differences in gender-predominant expression of organic anion transporter (Oat1-3; Slc22a6-8) mRNA levels. Drug Metab Dispos. 2004;32(6):620–5.CrossRefGoogle Scholar
  19. 19.
    Chen C, Klaassen CD. Rat multidrug resistance protein 4 (Mrp4, Abcc4): molecular cloning, organ distribution, postnatal renal expression, and chemical inducibility. Biochem Biophys Res Commun. 2004;317(1):46–53.CrossRefGoogle Scholar
  20. 20.
    Maher JM, Slitt AL, Cherrington NJ, Cheng X, Klaassen CD. Tissue distribution and hepatic and renal ontogeny of the multidrug resistance-associated protein (Mrp) family in mice. Drug Metab Dispos. 2005;33(7):947–55.CrossRefGoogle Scholar
  21. 21.
    Nakajima N, Sekine T, Cha SH, Tojo A, Hosoyamada M, Kanai Y, et al. Developmental changes in multispecific organic anion transporter 1 expression in the rat kidney. Kidney Int. 2000;57(4):1608–16.CrossRefGoogle Scholar
  22. 22.
    Kavlock RJ, Gray JA. Evaluation of renal function in neonatal rats. Biol Neonate. 1982;41(5–6):279–88.CrossRefGoogle Scholar
  23. 23.
    Ohtsuki S, Schaefer O, Kawakami H, Inoue T, Liehner S, Saito A, et al. Simultaneous absolute protein quantification of transporters, cytochromes P450, and UDP-glucuronosyltransferases as a novel approach for the characterization of individual human liver: comparison with mRNA levels and activities. Drug Metab Dispos. 2012;40(1):83–92.CrossRefGoogle Scholar
  24. 24.
    Slitt AL, Cherrington NJ, Hartley DP, Leazer TM, Klaassen CD. Tissue distribution and renal developmental changes in rat organic cation transporter mRNA levels. Drug Metab Dispos. 2002;30(2):212–9.CrossRefGoogle Scholar
  25. 25.
    Nozaki Y, Kusuhara H, Kondo T, Hasegawa M, Shiroyanagi Y, Nakazawa H, et al. Characterization of the uptake of organic anion transporter (OAT) 1 and OAT3 substrates by human kidney slices. J Pharmacol Exp Ther. 2007;321(1):362–9.CrossRefGoogle Scholar
  26. 26.
    Barter ZE, Perrett HF, Yeo KR, Allorge D, Lennard MS, Rostami-Hodjegan A. Determination of a quantitative relationship between hepatic CYP3A5*1/*3 and CYP3A4 expression for use in the prediction of metabolic clearance in virtual populations. Biopharm Drug Dispos. 2010;31(8–9):516–32.CrossRefGoogle Scholar

Copyright information

© American Association of Pharmaceutical Scientists 2019

Authors and Affiliations

  • Cindy Yanfei Li
    • 1
  • Chelsea Hosey-Cojocari
    • 2
  • Abdul Basit
    • 1
  • Jashvant D. Unadkat
    • 1
  • J. Steven Leeder
    • 2
  • Bhagwat Prasad
    • 1
    Email author
  1. 1.Department of PharmaceuticsUniversity of WashingtonSeattleUSA
  2. 2.Children’s Mercy Hospital and ClinicsKansas CityUSA

Personalised recommendations