Review Article

Advances in predictive in vitro models of drug-induced nephrotoxicity

Published online:

Abstract

In vitro screens for nephrotoxicity are currently poorly predictive of toxicity in humans. Although the functional proteins that are expressed by nephron tubules and mediate drug susceptibility are well known, current in vitro cellular models poorly replicate both the morphology and the function of kidney tubules and therefore fail to demonstrate injury responses to drugs that would be nephrotoxic in vivo. Advances in protocols to enable the directed differentiation of pluripotent stem cells into multiple renal cell types and the development of microfluidic and 3D culture systems have opened a range of potential new platforms for evaluating drug nephrotoxicity. Many of the new in vitro culture systems have been characterized by the expression and function of transporters, enzymes, and other functional proteins that are expressed by the kidney and have been implicated in drug-induced renal injury. In vitro platforms that express these proteins and exhibit molecular biomarkers that have been used as readouts of injury demonstrate improved functional maturity compared with static 2D cultures and represent an opportunity to model injury to renal cell types that have hitherto received little attention. As nephrotoxicity screening platforms become more physiologically relevant, they will facilitate the development of safer drugs and improved clinical management of nephrotoxicants.

Key points

  • Currently available in vitro and animal models of drug-induced nephrotoxicity are poorly predictive of toxicity in humans.

  • Functional proteins that underlie the susceptibility of various renal cell types to specific drugs, and molecular biomarkers of injury, can be used to characterize the functional maturity of in vitro models and their capacity to respond to nephrotoxicants.

  • In vitro models derived using new protocols for the directed differentiation of pluripotent stem cells to renal cells and new 3D in vitro culture systems demonstrate improved functional maturity over static 2D systems.

  • Improved functional maturity of cultured renal cells in systems that more closely replicate the physiology of the renal tubule and its supporting cells will improve the predictive ability of in vitro models of nephrotoxicity.

  • Subscribe to Nature Reviews Nephrology for full access:

    $199

    Subscribe

Additional access options:

Already a subscriber?  Log in  now or  Register  for online access.

Additional information

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

GUDMAP:www.gudmap.org

Regenerative Medicine Crossing Borders:www.regmedxb.com

References

  1. 1.

    Nolin, T. D. & Himmelfarb, J. in Adverse Drug Reactions (ed. Uetrecht, J.) 111–130 (Springer, Berlin, Heidelberg, 2010).

  2. 2.

    Grünfeld, J.-P. & Rossier, B. C. Lithium nephrotoxicity revisited. Nat. Rev. Nephrol. 5, 270–276 (2009).

  3. 3.

    Rewa, O. & Bagshaw, S. M. Acute kidney injury — epidemiology, outcomes and economics. Nat. Rev. Nephrol. 10, 193–207 (2014).

  4. 4.

    Magee, T. V. et al. Discovery of Dap-3 polymyxin analogues for the treatment of multidrug-resistant Gram-negative nosocomial infections. J. Med. Chem. 56, 5079–5093 (2013).

  5. 5.

    Tiong, H. Y. et al. Drug-induced nephrotoxicity: clinical impact and preclinical in vitro models. Mol. Pharm 11, 1933–1948 (2014).

  6. 6.

    Huang, J. X. et al. Evaluation of biomarkers for in vitro prediction of drug-induced nephrotoxicity: comparison of HK-2, immortalized human proximal tubule epithelial, and primary cultures of human proximal tubular cells. Pharmacol. Res. Perspect. 3, e00148 (2015).

  7. 7.

    Dekant, W. & Vamvakas, S. Biotransformation and membrane transport in nephrotoxicity. Crit. Rev. Toxicol. 26, 309–334 (1996).

  8. 8.

    Hawksworth, G. M. et al. in Toxicology — From Cells to Man (eds Seiler, S. P., Kroftová, O. & Eybl, V.) 184–192 (Springer, Berlin, Heidelberg, 1996).

  9. 9.

    Lock, E. A. & Reed, C. J. Xenobiotic metabolizing enzymes of the kidney. Toxicol. Pathol. 26, 18–25 (1998).

  10. 10.

    Knops, N. et al. The functional implications of common genetic variation in CYP3A5 and ABCB1 in human proximal tubule cells. Mol. Pharm. 12, 758–768 (2015).

  11. 11.

    Bhargava, P. & Schnellmann, R. G. Mitochondrial energetics in the kidney. Nat. Rev. Nephrol. 13, 629–646 (2017).

  12. 12.

    Kramann, R., Tanaka, M. & Humphreys, B. D. Fluorescence microangiography for quantitative assessment of peritubular capillary changes after AKI in mice. J. Am. Soc. Nephrol. 25, 1924–1931 (2014).

  13. 13.

    Qi, W., Johnson, D. W., Vesey, D. A., Pollock, C. A. & Chen, X. Isolation, propagation and characterization of primary tubule cell culture from human kidney (Methods in Renal Research). Nephrology 12, 155–159 (2007).

  14. 14.

    Fisel, P., Renner, O., Nies, A. T., Schwab, M. & Schaeffeler, E. Solute carrier transporter and drug-related nephrotoxicity: the impact of proximal tubule cell models for preclinical research. Expert Opin. Drug Metab. Toxicol. 10, 395–408 (2014).

  15. 15.

    Lemke, A., Kiderlen, A. F. & Kayser, O. Amphotericin, B. Appl. Microbiol. Biotechnol. 68, 151–162 (2005).

  16. 16.

    Mamoulakis, C. et al. Contrast-induced nephropathy: Basic concepts, pathophysiological implications and prevention strategies. Pharmacol. Ther. 180, 99–112 (2017).

  17. 17.

    Paueksakon, P. & Fogo, A. B. Drug-induced nephropathies. Histopathology 70, 94–108 (2017).

  18. 18.

    Perazella, M. A. & Markowitz, G. S. Bisphosphonate nephrotoxicity. Kidney Int. 74, 1385–1393 (2008).

  19. 19.

    Xia, L., Zhou, M., Kalhorn, T. F., Ho, H. T. B. & Wang, J. Podocyte-specific expression of organic cation transporter PMAT: implication in puromycin aminonucleoside nephrotoxicity. Am. J. Physiol.-Ren. Physiol. 296, F1307–F1313 (2009).

  20. 20.

    Yilmaz, M., Taninmis, H., Kara, E., Ozagari, A. & Unsal, A. Nephrotic syndrome after oral bisphosphonate (alendronate) administration in a patient with osteoporosis. Osteoporos. Int. 23, 2059–2062 (2012).

  21. 21.

    Cheng, H. F. & Harris, R. C. Renal effects of non-steroidal anti-inflammatory drugs and selective cyclooxygenase-2 inhibitors. Curr. Pharm. Des. 11, 1795–1804 (2005).

  22. 22.

    Perazella, M. A. Drug-induced renal failure: update on new medications and unique mechanisms of nephrotoxicity. Am. J. Med. Sci. 325, 349–362 (2003).

  23. 23.

    Basile, D. P., Donohoe, D., Roethe, K. & Osborn, J. L. Renal ischemic injury results in permanent damage to peritubular capillaries and influences long-term function. Am. J. Physiol. Renal Physiol. 281, F887–F899 (2001).

  24. 24.

    Verma, S. K. & Molitoris, B. A. Renal endothelial injury and microvascular dysfunction in acute kidney injury. Semin. Nephrol. 35, 96–107 (2015).

  25. 25.

    Dimke, H. et al. Tubulovascular cross-talk by vascular endothelial growth factor a maintains peritubular microvasculature in kidney. J. Am. Soc. Nephrol. 26, 1027–1038 (2015).

  26. 26.

    Lameire, N. Nephrotoxicity of recent anti-cancer agents. Clin. Kidney J. 7, 11–22 (2014).

  27. 27.

    Al-Nouri, Z. L., Reese, J. A., Terrell, D. R., Vesely, S. K. & George, J. N. Drug-induced thrombotic microangiopathy: a systematic review of published reports. Blood 125, 616–618 (2015).

  28. 28.

    Breljak, D. et al. Distribution of organic anion transporters NaDC3 and OAT1-3 along the human nephron. Am. J. Physiol. Renal Physiol. 311, F227–F238 (2016).

  29. 29.

    Motohashi, H. et al. Gene expression levels and immunolocalization of organic ion transporters in the human kidney. J. Am. Soc. Nephrol. 13, 866–874 (2002).

  30. 30.

    Ingraham, L. et al. A plasma concentration of α-ketoglutarate influences the kinetic interaction of ligands with organic anion transporter 1. Mol. Pharmacol. 86, 86–95 (2014).

  31. 31.

    Budiman, T., Bamberg, E., Koepsell, H. & Nagel, G. Mechanism of electrogenic cation transport by the cloned organic cation transporter 2 from rat. J. Biol. Chem. 275, 29413–29420 (2000).

  32. 32.

    Cihlar, T. et al. The antiviral nucleotide analogs cidofovir and adefovir are novel substrates for human and rat renal organic anion transporter 1. Mol. Pharmacol. 56, 570–580 (1999).

  33. 33.

    Ho, E. S., Lin, D. C., Mendel, D. B. & Cihlar, T. Cytotoxicity of antiviral nucleotides adefovir and cidofovir is induced by the expression of human renal organic anion transporter 1. J. Am. Soc. Nephrol. 11, 383–393 (2000).

  34. 34.

    Nieskens, T. T. G. et al. A human renal proximal tubule cell line with stable organic anion transporter 1 and 3 expression predictive for antiviral-induced toxicity. AAPS J. 18, 465–475 (2016).

  35. 35.

    Hagos, Y. & Wolff, N. A. Assessment of the role of renal organic anion transporters in drug-induced nephrotoxicity. Toxins 2, 2055–2082 (2010).

  36. 36.

    Ciarimboli, G. Role of organic cation transporters in drug-induced toxicity. Expert Opin. Drug Metab. Toxicol. 7, 159–174 (2011).

  37. 37.

    Pabla, N., Murphy, R. F., Liu, K. & Dong, Z. The copper transporter Ctr1 contributes to cisplatin uptake by renal tubular cells during cisplatin nephrotoxicity. Am. J. Physiol. Renal Physiol. 296, F505–F511 (2009).

  38. 38.

    Hori, Y. et al. Megalin blockade with cilastatin suppresses drug-induced nephrotoxicity. J. Am. Soc. Nephrol. 28, 1783–1791 (2017).

  39. 39.

    Nagai, J. & Takano, M. Entry of aminoglycosides into renal tubular epithelial cells via endocytosis-dependent and endocytosis-independent pathways. Biochem. Pharmacol. 90, 331–337 (2014).

  40. 40.

    [No authors listed.] Genitourinary Development Molecular Anatomy Project. GUDMAP http://www.gudmap.org/ (2017).

  41. 41.

    Harding, S. D. et al. The GUDMAP database — an online resource for genitourinary research. Development 138, 2845–2853 (2011).

  42. 42.

    Jenkinson, S. E. et al. The limitations of renal epithelial cell line HK-2 as a model of drug transporter expression and function in the proximal tubule. Pflüg. Arch. 464, 601–611 (2012).

  43. 43.

    van Aubel, R. A., Smeets, P. H., Peters, J. G., Bindels, R. J. & Russel, F. G. The MRP4/ABCC4 gene encodes a novel apical organic anion transporter in human kidney proximal tubules: putative efflux pump for urinary cAMP and cGMP. J. Am. Soc. Nephrol. 13, 595–603 (2002).

  44. 44.

    Huls, M. et al. The breast cancer resistance protein transporter ABCG2 is expressed in the human kidney proximal tubule apical membrane. Kidney Int. 73, 220–225 (2008).

  45. 45.

    Motohashi, H. & Inui, K. Multidrug and toxin extrusion family SLC47: Physiological, pharmacokinetic and toxicokinetic importance of MATE1 and MATE2-K. Mol. Aspects Med. 34, 661–668 (2013).

  46. 46.

    Wen, X. et al. MDR1 transporter protects against paraquat-induced toxicity in human and mouse proximal tubule cells. Toxicol. Sci. 141, 475–483 (2014).

  47. 47.

    Yokoo, S. et al. Differential contribution of organic cation transporters, OCT2 and MATE1, in platinum agent-induced nephrotoxicity. Biochem. Pharmacol. 74, 477–487 (2007).

  48. 48.

    Hahn, K., Ejaz, A. A., Kanbay, M., Lanaspa, M. A. & Johnson, R. J. Acute kidney injury from SGLT2 inhibitors: potential mechanisms. Nat. Rev. Nephrol. 12, 711–712 (2016).

  49. 49.

    Nadkarni, G. N. et al. Acute kidney injury in patients on SGLT2 inhibitors: a propensity-matched analysis. Diabetes Care 40, 1479–1485 (2017).

  50. 50.

    Saly, D. & Perazella, M. A. Harnessing basic and clinic tools to evaluate SGLT2 inhibitor nephrotoxicity. Am. J. Physiol. Renal Physiol. 313, F951–F954 (2017).

  51. 51.

    Yu, A. S. L. Claudins and the kidney. J. Am. Soc. Nephrol. 26, 11–19 (2015).

  52. 52.

    Markadieu, N. et al. A primary culture of distal convoluted tubules expressing functional thiazide-sensitive NaCl transport. Am. J. Physiol. Renal Physiol. 303, F886–F892 (2012).

  53. 53.

    Fromm, M., Piontek, J., Rosenthal, R., Günzel, D. & Krug, S. M. Tight junctions of the proximal tubule and their channel proteins. Pflüg. Arch. 469, 877–887 (2017).

  54. 54.

    Günzel, D. & Yu, A. S. L. Claudins and the modulation of tight junction permeability. Physiol. Rev. 93, 525–569 (2013).

  55. 55.

    Trujillo, J. et al. Renal tight junction proteins are decreased in cisplatin-induced nephrotoxicity in rats. Toxicol. Mech. Methods 24, 520–528 (2014).

  56. 56.

    Balkovetz, D. F. Tight junction claudins and the kidney in sickness and in health. Biochim. Biophys. Acta 1788, 858–863 (2009).

  57. 57.

    Lash, L. H., Putt, D. A. & Cai, H. Drug metabolism enzyme expression and activity in primary cultures of human proximal tubular cells. Toxicology 244, 56–65 (2008).

  58. 58.

    Miners, J., Yang, X., Knights, K. & Zhang, L. The role of the kidney in drug elimination: transport, metabolism, and the impact of kidney disease on drug clearance. Clin. Pharmacol. Ther. 102, 436–449 (2017).

  59. 59.

    Bolbrinker, J. et al. CYP3A5 genotype-phenotype analysis in the human kidney reveals a strong site-specific expression of CYP3A5 in the proximal tubule in carriers of the CYP3A5*1 allele. Drug Metab. Dispos. 40, 639–641 (2012).

  60. 60.

    Yu, J., Zhou, Z., Owens, K. H., Ritchie, T. K. & Ragueneau-Majlessi, I. What can be learned from recent new drug applications? A systematic review of drug interaction data for drugs approved by the US FDA in 2015. Drug Metab. Dispos. 45, 86–108 (2017).

  61. 61.

    Dekant, W. The role of biotransformation and bioactivation in toxicity. EXS 99, 57–86 (2009).

  62. 62.

    Liu, S. et al. The role of renal proximal tubule P450 enzymes in chloroform-induced nephrotoxicity: Utility of renal specific P450 reductase knockout mouse models. Toxicol. Appl. Pharmacol. 272, 230–237 (2013).

  63. 63.

    Fliedl, L. Controversial role of gamma-glutamyl transferase activity in cisplatin nephrotoxicity. ALTEX 31, 269–278 (2014).

  64. 64.

    Meister, A., Tate, S. S. & Griffith, O. W. Gamma-glutamyl transpeptidase. Methods Enzymol. 77, 237–253 (1981).

  65. 65.

    King, S. M. et al. 3D proximal tubule tissues recapitulate key aspects of renal physiology to enable nephrotoxicity testing. Front. Physiol. 8, 123 (2017).

  66. 66.

    Wieser, M. et al. hTERT alone immortalizes epithelial cells of renal proximal tubules without changing their functional characteristics. Am. J. Physiol. Renal Physiol. 295, F1365–F1375 (2008).

  67. 67.

    Weber, E. J. et al. Development of a microphysiological model of human kidney proximal tubule function. Kidney Int. 90, 627–637 (2016).

  68. 68.

    Kortenoeven, M. L. A. et al. Amiloride blocks lithium entry through the sodium channel thereby attenuating the resultant nephrogenic diabetes insipidus. Kidney Int. 76, 44–53 (2009).

  69. 69.

    Thomsen, K. & Shirley, D. G. A hypothesis linking sodium and lithium reabsorption in the distal nephron. Nephrol. Dial. Transplant. 21, 869–880 (2006).

  70. 70.

    Christensen, B. M., Kim, Y.-H., Kwon, T.-H. & Nielsen, S. Lithium treatment induces a marked proliferation of primarily principal cells in rat kidney inner medullary collecting duct. Am. J. Physiol. Renal Physiol. 291, F39–F48 (2006).

  71. 71.

    Ledeganck, K. J. et al. Expression of renal distal tubule transporters TRPM6 and NCC in a rat model of cyclosporine nephrotoxicity and effect of EGF treatment. Am. J. Physiol. Renal Physiol. 301, F486–F493 (2011).

  72. 72.

    Bonventre, J. V., Vaidya, V. S., Schmouder, R., Feig, P. & Dieterle, F. Next-generation biomarkers for detecting kidney toxicity. Nat. Biotechnol. 28, 436–440 (2010).

  73. 73.

    Bailly, V. et al. Shedding of kidney injury molecule-1, a putative adhesion protein involved in renal regeneration. J. Biol. Chem. 277, 39739–39748 (2002).

  74. 74.

    Ichimura, T. Kidney injury molecule-1: a tissue and urinary biomarker for nephrotoxicant-induced renal injury. Am. J. Physiol. Renal Physiol. 286, 552F–563 (2004).

  75. 75.

    Ichimura, T. et al. Kidney injury molecule-1 is a phosphatidylserine receptor that confers a phagocytic phenotype on epithelial cells. J. Clin. Invest. 118, 1657–1668 (2008).

  76. 76.

    Vaidya, V. S. Urinary kidney injury molecule-1: a sensitive quantitative biomarker for early detection of kidney tubular injury. Am. J. Physiol. Renal Physiol. 290, F517–F529 (2006).

  77. 77.

    Vaidya, V. S. et al. Kidney injury molecule-1 outperforms traditional biomarkers of kidney injury in preclinical biomarker qualification studies. Nat. Biotechnol. 28, 478–485 (2010).

  78. 78.

    Vaidya, V. S. et al. Urinary biomarkers for sensitive and specific detection of acute kidney injury in humans. Clin. Transl Sci. 1, 200–208 (2008).

  79. 79.

    Dieterle, F. et al. Renal biomarker qualification submission: a dialog between the FDA-EMEA and Predictive Safety Testing Consortium. Nat. Biotechnol. 28, 455–462 (2010).

  80. 80.

    Rached, E. et al. Evaluation of putative biomarkers of nephrotoxicity after exposure to ochratoxin A in vivo and in vitro. Toxicol. Sci 103, 371–381 (2008).

  81. 81.

    Sohn, S.-J. et al. In vitro evaluation of biomarkers for cisplatin-induced nephrotoxicity using HK-2 human kidney epithelial cells. Toxicol. Lett. 217, 235–242 (2013).

  82. 82.

    Li, Y. et al. An in vitro method for the prediction of renal proximal tubular toxicity in humans. Toxicol. Res 2, 352 (2013).

  83. 83.

    Luo, Q.-H. et al. Evaluation of KIM-1 and NGAL as early indicators for assessment of gentamycin-induced nephrotoxicity in vivo and in vitro. Kidney Blood Press. Res 41, 911–918 (2016).

  84. 84.

    Gozzelino, R., Jeney, V. & Soares, M. P. Mechanisms of cell protection by heme oxygenase-1. Annu. Rev. Pharmacol. Toxicol. 50, 323–354 (2010).

  85. 85.

    Lever, J. M., Boddu, R., George, J. F. & Agarwal, A. Heme oxygenase-1 in kidney health and disease. Antioxid. Redox Signal. https://doi.org/10.1089/ars.2016.6659 (2016).

  86. 86.

    Zager, R. A., Johnson, A. C. M. & Becker, K. Plasma and urinary heme oxygenase-1 in AKI. J. Am. Soc. Nephrol. 23, 1048–1057 (2012).

  87. 87.

    Adler, M. et al. A quantitative approach to screen for nephrotoxic compounds in vitro. J. Am. Soc. Nephrol. 27, 1015–1028 (2016).

  88. 88.

    Akdis, M. et al. Interleukins, from 1 to 37, and interferon-γ: receptors, functions, and roles in diseases. J. Allergy Clin. Immunol. 127, 701–721.e70 (2011).

  89. 89.

    Tramma, D., Hatzistylianou, M., Gerasimou, G. & Lafazanis, V. Interleukin-6 and interleukin-8 levels in the urine of children with renal scarring. Pediatr. Nephrol. 27, 1525–1530 (2012).

  90. 90.

    Grigoryev, D. N. et al. The local and systemic inflammatory transcriptome after acute kidney injury. J. Am. Soc. Nephrol. 19, 547–558 (2008).

  91. 91.

    Araki, M. et al. Expression of IL-8 during reperfusion of renal allografts is dependent on ischemic time. Transplantation 81, 783–788 (2006).

  92. 92.

    Su, H., Lei, C.-T. & Zhang, C. Interleukin-6 signaling pathway and its role in kidney disease: an update. Front. Immunol. 8, 405 (2017).

  93. 93.

    Mihajlovic, M. et al. Allostimulatory capacity of conditionally immortalized proximal tubule cell lines for bioartificial kidney application. Sci. Rep. 7, 7103 (2017).

  94. 94.

    Cowland, J. B. & Borregaard, N. Molecular characterization and pattern of tissue expression of the gene for neutrophil gelatinase-associated lipocalin from humans. Genomics 45, 17–23 (1997).

  95. 95.

    Charlton, J. R., Portilla, D. & Okusa, M. D. A basic science view of acute kidney injury biomarkers. Nephrol. Dial. Transplant. 29, 1301–1311 (2014).

  96. 96.

    Paragas, N. et al. The Ngal reporter mouse detects the response of the kidney to injury in real time. Nat. Med. 17, 216–222 (2011).

  97. 97.

    McIlroy, D. R., Wagener, G. & Lee, H. T. Neutrophil gelatinase-associated lipocalin and acute kidney injury after cardiac surgery: the effect of baseline renal function on diagnostic performance. Clin. J. Am. Soc. Nephrol. 5, 211–219 (2010).

  98. 98.

    Mårtensson, J. & Bellomo, R. The rise and fall of NGAL in acute kidney injury. Blood Purif. 37, 304–310 (2014).

  99. 99.

    Haase, M. et al. Accuracy of neutrophil gelatinase-associated lipocalin (NGAL) in diagnosis and prognosis in acute kidney injury: a systematic review and meta-analysis. Am. J. Kidney Dis. 54, 1012–1024 (2009).

  100. 100.

    Scotcher, D., Jones, C., Posada, M., Galetin, A. & Rostami-Hodjegan, A. Key to opening kidney for in vitro-in vivo extrapolation entrance in health and disease: Part II: mechanistic models and in vitro-in vivo extrapolation. AAPS J. 18, 1082–1094 (2016).

  101. 101.

    Hilgendorf, C. et al. Expression of thirty-six drug transporter genes in human intestine, liver, kidney, and organotypic cell lines. Drug Metab. Dispos. 35, 1333–1340 (2007).

  102. 102.

    Chu, X., Bleasby, K. & Evers, R. Species differences in drug transporters and implications for translating preclinical findings to humans. Expert Opin. Drug Metab. Toxicol. 9, 237–252 (2013).

  103. 103.

    Yonezawa, A. & Inui, K. Importance of the multidrug and toxin extrusion MATE/SLC47A family to pharmacokinetics, pharmacodynamics/toxicodynamics and pharmacogenomics. Br. J. Pharmacol. 164, 1817–1825 (2011).

  104. 104.

    Aoki, M. et al. Kidney-specific expression of human organic cation transporter 2 (OCT2/SLC22A2) is regulated by DNA methylation. Am. J. Physiol. Renal Physiol. 295, F165–F170 (2008).

  105. 105.

    Tanaka, Y., Slitt, A. L., Leazer, T. M., Maher, J. M. & Klaassen, C. D. Tissue distribution and hormonal regulation of the breast cancer resistance protein (Bcrp/Abcg2) in rats and mice. Biochem. Biophys. Res. Commun. 326, 181–187 (2004).

  106. 106.

    Soldin, O. P. & Mattison, D. R. Sex differences in pharmacokinetics and pharmacodynamics. Clin. Pharmacokinet. 48, 143–157 (2009).

  107. 107.

    Joseph, S. et al. Expression of drug transporters in human kidney: impact of sex, age, and ethnicity. Biol. Sex Differ. 6, 4 (2015).

  108. 108.

    Veiras, L. C. et al. Sexual dimorphic pattern of renal transporters and electrolyte homeostasis. J. Am. Soc. Nephrol. 28, 3504–3517 (2017).

  109. 109.

    [No authors listed.] Validating human stem cell cardiomyocyte technology for better predictive assessment of drug-induced cardiac toxicity. U.S. Food & Drug Administration https://www.fda.gov/ScienceResearch/SpecialTopics/RegulatoryScience/ucm507998.htm (2016).

  110. 110.

    Wilmer, M. J. et al. Kidney-on-a-chip technology for drug-induced nephrotoxicity screening. Trends Biotechnol. 34, 156–170 (2016).

  111. 111.

    Ryan, M. J. et al. HK-2: An immortalized proximal tubule epithelial cell line from normal adult human kidney. Kidney Int. 45, 48–57 (1994).

  112. 112.

    Wilmer, M. J. et al. Novel conditionally immortalized human proximal tubule cell line expressing functional influx and efflux transporters. Cell Tissue Res. 339, 449–457 (2010).

  113. 113.

    Aschauer, L., Carta, G., Vogelsang, N., Schlatter, E. & Jennings, P. Expression of xenobiotic transporters in the human renal proximal tubule cell line RPTEC/TERT1. Toxicol. In Vitro 30, 95–105 (2015).

  114. 114.

    Schophuizen, C. M. S. et al. Cationic uremic toxins affect human renal proximal tubule cell functioning through interaction with the organic cation transporter. Pflüg. Arch. 465, 1701–1714 (2013).

  115. 115.

    Mutsaers, H. A. M. et al. Uremic toxins inhibit renal metabolic capacity through interference with glucuronidation and mitochondrial respiration. Biochim. Biophys. Acta 1832, 142–150 (2013).

  116. 116.

    Jansen, J. et al. Bioengineered kidney tubules efficiently excrete uremic toxins. Sci. Rep. 6, 26715 (2016).

  117. 117.

    Ivliev, A. E., ’t Hoen, P. A. C., Roon-Mom, W. M. C., van, Peters, D. J. M. & Sergeeva, M. G. Exploring the transcriptome of ciliated cells using in silico dissection of human tissues. PLoS ONE 7, e35618 (2012).

  118. 118.

    Ohnuki, M. & Takahashi, K. Present and future challenges of induced pluripotent stem cells. Phil. Trans. R. Soc. B Biol Sci. 370, 20140367 (2015).

  119. 119.

    Lam, A. Q. et al. Rapid and efficient differentiation of human pluripotent stem cells into intermediate mesoderm that forms tubules expressing kidney proximal tubular markers. J. Am. Soc. Nephrol. 25, 1211–1225 (2014).

  120. 120.

    Mae, S.-I. et al. Monitoring and robust induction of nephrogenic intermediate mesoderm from human pluripotent stem cells. Nat. Commun. 4, 1367 (2013).

  121. 121.

    Xia, Y. et al. The generation of kidney organoids by differentiation of human pluripotent cells to ureteric bud progenitor-like cells. Nat. Protoc. 9, 2693–2704 (2014).

  122. 122.

    Taguchi, A. et al. Redefining the in vivo origin of metanephric nephron progenitors enables generation of complex kidney structures from pluripotent stem cells. Cell Stem Cell 14, 53–67 (2014).

  123. 123.

    Freedman, B. S. et al. Modelling kidney disease with CRISPR-mutant kidney organoids derived from human pluripotent epiblast spheroids. Nat. Commun. 6, 8715 (2015).

  124. 124.

    Takasato, M. et al. Kidney organoids from human iPS cells contain multiple lineages and model human nephrogenesis. Nature 526, 564–568 (2015).

  125. 125.

    Morizane, R. et al. Nephron organoids derived from human pluripotent stem cells model kidney development and injury. Nat. Biotechnol. 33, 1193–1200 (2015).

  126. 126.

    Kandasamy, K. et al. Prediction of drug-induced nephrotoxicity and injury mechanisms with human induced pluripotent stem cell-derived cells and machine learning methods. Sci. Rep. 5, 12337 (2015).

  127. 127.

    Imberti, B. et al. Renal progenitors derived from human iPSCs engraft and restore function in a mouse model of acute kidney injury. Sci. Rep. 5, 8826 (2015).

  128. 128.

    Musah, S. et al. Mature induced-pluripotent-stem-cell-derived human podocytes reconstitute kidney glomerular-capillary-wall function on a chip. Nat. Biomed. Eng. 1, 0069 (2017).

  129. 129.

    Song, B. et al. The directed differentiation of human iPS cells into kidney podocytes. PLoS ONE 7, e46453 (2012).

  130. 130.

    Kaminski, M. M. et al. Direct reprogramming of fibroblasts into renal tubular epithelial cells by defined transcription factors. Nat. Cell Biol. 18, 1269–1280 (2016).

  131. 131.

    Xu, J., Du, Y. & Deng, H. Direct lineage reprogramming: strategies, mechanisms, and applications. Cell Stem Cell 16, 119–134 (2015).

  132. 132.

    Takasato, M. et al. Directing human embryonic stem cell differentiation towards a renal lineage generates a self-organizing kidney. Nat. Cell Biol. 16, 118–126 (2014).

  133. 133.

    Jang, K.-J. et al. Human kidney proximal tubule-on-a-chip for drug transport and nephrotoxicity assessment. Integr. Biol. 5, 1119 (2013).

  134. 134.

    Homan, K. A. et al. Bioprinting of 3D convoluted renal proximal tubules on perfusable chips. Sci. Rep. 6, 34845 (2016).

  135. 135.

    Tourovskaia, A., Fauver, M., Kramer, G., Simonson, S. & Neumann, T. Tissue-engineered microenvironment systems for modeling human vasculature. Exp. Biol. Med. 239, 1264–1271 (2014).

  136. 136.

    Masereeuw, R. et al. Probenecid interferes with renal oxidative metabolism: a potential pitfall in its use as an inhibitor of drug transport. Br. J. Pharmacol. 131, 57–62 (2000).

  137. 137.

    Tsai, M. et al. In vitro modeling of the microvascular occlusion and thrombosis that occur in hematologic diseases using microfluidic technology. J. Clin. Invest. 122, 408–418 (2012).

  138. 138.

    Ligresti, G. et al. A novel three-dimensional human peritubular microvascular system. J. Am. Soc. Nephrol. 27, 2370–2381 (2016).

  139. 139.

    Kelly, E. J. et al. Innovations in preclinical biology: ex vivo engineering of a human kidney tissue microperfusion system. Stem Cell Res. Ther. 4, S17 (2013).

  140. 140.

    Phan, D. T. T. et al. A vascularized and perfused organ-on-a-chip platform for large-scale drug screening applications. Lab Chip 17, 511–520 (2017).

  141. 141.

    van Duinen, V. et al. 96 perfusable blood vessels to study vascular permeability in vitro. Sci. Rep. 7, 18071 (2017).

  142. 142.

    Vernetti, L. et al. Functional coupling of human microphysiology systems: intestine, liver, kidney proximal tubule, blood-brain barrier and skeletal muscle. Sci. Rep. 7, 42296 (2017).

  143. 143.

    Chang, S.-Y. et al. Human liver-kidney model elucidates the mechanisms of aristolochic acid nephrotoxicity. JCI Insight 2, 95978 (2017).

  144. 144.

    Jansen, J. et al. Human proximal tubule epithelial cells cultured on hollow fibers: living membranes that actively transport organic cations. Sci. Rep. 5, 16702 (2015).

  145. 145.

    Mihajlovic, M. et al. Role of vitamin D in maintaining renal epithelial barrier function in uremic conditions. Int. J. Mol. Sci. 18, 2531 (2017).

  146. 146.

    Nguyen, D. G. et al. Bioprinted 3D primary liver tissues allow assessment of organ-level response to clinical drug induced toxicity in vitro. PLoS ONE 11, e0158674 (2016).

  147. 147.

    Persson, M. & Hornberg, J. J. Advances in predictive toxicology for discovery safety through high content screening. Chem. Res. Toxicol. 29, 1998–2007 (2016).

  148. 148.

    Su, R., Xiong, S., Zink, D. & Loo, L.-H. High-throughput imaging-based nephrotoxicity prediction for xenobiotics with diverse chemical structures. Arch. Toxicol. 90, 2793–2808 (2016).

  149. 149.

    Abdullah, R., Alhusainy, W., Woutersen, J., Rietjens, I. M. C. M. & Punt, A. Predicting points of departure for risk assessment based on in vitro cytotoxicity data and physiologically based kinetic (PBK) modeling: The case of kidney toxicity induced by aristolochic acid I. Food Chem. Toxicol. 92, 104–116 (2016).

  150. 150.

    Zhou, S.-F., Liu, J.-P. & Chowbay, B. Polymorphism of human cytochrome P450 enzymes and its clinical impact. Drug Metab. Rev. 41, 89–295 (2009).

  151. 151.

    Scotcher, D. et al. Microsomal and cytosolic scaling factors in dog and human kidney cortex and application for in vitro-in vivo extrapolation of renal metabolic clearance. Drug Metab. Dispos. 45, 556–568 (2017).

  152. 152.

    Leclerc, E., Hamon, J. & Bois, F. Y. Investigation of ifosfamide and chloroacetaldehyde renal toxicity through integration of in vitro liver–kidney microfluidic data and pharmacokinetic-system biology models. J. Appl. Toxicol. 36, 330–339 (2016).

  153. 153.

    Miller, R. P., Tadagavadi, R. K., Ramesh, G. & Reeves, W. B. Mechanisms of cisplatin nephrotoxicity. Toxins 2, 2490–2518 (2010).

  154. 154.

    Nakamura, T., Yonezawa, A., Hashimoto, S., Katsura, T. & Inui, K. Disruption of multidrug and toxin extrusion MATE1 potentiates cisplatin-induced nephrotoxicity. Biochem. Pharmacol. 80, 1762–1767 (2010).

  155. 155.

    Hu, S. et al. Identification of OAT1/OAT3 as contributors to cisplatin toxicity. Clin. Transl Sci. 10, 412–420 (2017).

  156. 156.

    Sonneveld, R. et al. Glucose specifically regulates TRPC6 expression in the podocyte in an AngII-dependent manner. Am. J. Pathol. 184, 1715–1726 (2014).

  157. 157.

    Ambrus, L. et al. Inhibition of TRPC6 by protein kinase C isoforms in cultured human podocytes. J. Cell. Mol. Med. 19, 2771–2779 (2015).

  158. 158.

    Eyre, J. et al. Statin-sensitive endocytosis of albumin by glomerular podocytes. Am. J. Physiol. Renal Physiol. 292, F674–F681 (2007).

  159. 159.

    Kido, Y., Matsson, P. & Giacomini, K. M. Profiling of a prescription drug library for potential renal drug–drug interactions mediated by the organic cation transporter 2. J. Med. Chem. 54, 4548–4558 (2011).

  160. 160.

    Caetano-Pinto, P. et al. Fluorescence-based transport assays revisited in a human renal proximal tubule cell line. Mol. Pharm. 13, 933–944 (2016).

  161. 161.

    Shaik, N., Giri, N., Pan, G. & Elmquist, W. F. P-Glycoprotein-mediated active efflux of the anti-HIV1 nucleoside abacavir limits cellular accumulation and brain distribution. Drug Metab. Dispos. 35, 2076–2085 (2007).

  162. 162.

    Kusuhara, H. et al. Effects of a MATE protein inhibitor, pyrimethamine, on the renal elimination of metformin at oral microdose and at therapeutic dose in healthy subjects. Clin. Pharmacol. Ther. 89, 837–844 (2011).

  163. 163.

    Ito, S. et al. Potent and specific inhibition of mMate1-mediated efflux of type I organic cations in the liver and kidney by pyrimethamine. J. Pharmacol. Exp. Ther. 333, 341–350 (2010).

  164. 164.

    Zhai, X. Y. et al. Cubilin- and megalin-mediated uptake of albumin in cultured proximal tubule cells of opossum kidney. Kidney Int. 58, 1523–1533 (2000).

  165. 165.

    Moreno, E. et al. Affinity-defining domains in the Na-Cl cotransporter: a different location for Cl and thiazide binding. J. Biol. Chem. 281, 17266–17275 (2006).

  166. 166.

    Andrukhova, O. et al. FGF23 promotes renal calcium reabsorption through the TRPV5 channel. EMBO J. 33, 229–246 (2014).

  167. 167.

    Masereeuw, R., Moons, M. M., Toomey, B. H., Russel, F. G. M. & Miller, D. S. Active lucifer yellow secretion in renal proximal tubule: evidence for organic anion transport system crossover. J. Pharmacol. Exp. Ther. 289, 1104–1111 (1999).

Download references

Acknowledgements

J.Y.-C.S. is supported by a University of Melbourne Research Scholarship and a Murdoch Children’s Research Institute Top up scholarship. J.J. is supported by the Dutch Kidney Foundation (Kolff postdoctoral fellowship abroad grant 170KK05), EMBO (short-term fellowship 6893), and by the partners of Regenerative Medicine Crossing Borders powered by Health~Holland, Top Sector Life Sciences & Health. R.M. is supported by a grant from the UK National Centre for the Replacement, Refinement and Reduction of Animals in Research (NC3Rs), the NephroTube CRACK IT Challenge, and the RegMed XB consortium. M.H.L. is an Australian National Health and Medical Research Council (NHMRC) Senior Principal Research Fellow (GNT1042093) and is supported by funding from the NHMRC (GNT1100970) and the US National Institutes of Health (DK107344).

Reviewer information

Nature Reviews Nephrology thanks J. Lewis, R. Morizane and the other, anonymous reviewer(s) for their contribution to the peer review of this work.

Author information

Author notes

  1. These authors contributed equally: Joanne Y-C Soo, Jitske Jansen

Affiliations

  1. Department of Paediatrics, The University of Melbourne, Parkville, Victoria, Australia

    • Joanne Y.-C. Soo
    •  & Melissa H. Little
  2. Murdoch Children’s Research Institute, Parkville, Victoria, Australia

    • Joanne Y.-C. Soo
    •  & Melissa H. Little
  3. Division of Pharmacology, Utrecht Institute for Pharmaceutical Sciences, Utrecht University, Utrecht, Netherlands

    • Jitske Jansen
    •  & Rosalinde Masereeuw
  4. Department of Anatomy and Neuroscience, The University of Melbourne, Parkville, Victoria, Australia

    • Melissa H. Little

Authors

  1. Search for Joanne Y.-C. Soo in:

  2. Search for Jitske Jansen in:

  3. Search for Rosalinde Masereeuw in:

  4. Search for Melissa H. Little in:

Contributions

J.J. researched data for the article. J.Y.-C.S., J.J., and R.M. wrote the article. All authors contributed substantially to discussion of the article’s content and reviewed and edited the manuscript before submission.

Competing interests

M.H.L. holds a research contract with Organovo, Inc. The other authors declare no competing interests.

Corresponding author

Correspondence to Melissa H. Little.

Glossary

Nephrotoxicants

Any compounds, natural and synthetic, that exert an adverse effect on a specific kidney cell type or mediate an unwanted event affecting kidney functioning. By contrast, a toxin is a poisonous substance produced within living cells or organisms.

Electrogenic transport

Transport that leads to a change in net charge across a cell membrane.

Transepithelial electrical resistance

(TEER). The electrical resistance across a cell monolayer. The higher the value, the less permeable the monolayer.

Kidney-on-a-chip

Renal cells seeded in a 2D or 3D configuration in a microfluidic device. For proximal tubule chips, these devices typically allow flow of media across the cells’ apical surface, basolateral surface, or both. Other cell types such as endothelial cells might be included; in these cases, the organization of the different cell types is defined by the design of the chip.

Kidney organoids

Three-dimensional aggregates of interstitial cells and nephron structures with characteristic segments, typically formed by directing pluripotent stem cells to a renal fate and aggregating these cells to enable self-organization, with or without additional extracellular matrix.

Renal tissue arrays

Three-dimensional co-cultures of renal epithelial cells, renal fibroblasts, and endothelial cells. Cell suspensions are prepared in biocompatible gels and bioprinted. The composition of the suspensions and the spatial arrangement of the different suspensions used define the organization of the different cell types. Scaffolds composed of extracellular matrix may or may not be used.

Fugitive ink

Biocompatible material that can be printed and later evacuated to leave a hollow space within a mould.

Biofunctionalized hollow fibres

Hollow, porous polymer fibres coated with extracellular matrix on which renal cells can be cultured.

Desquamated

Peeling and shedding of the top layer of an epithelium.