Abstract
Drug-induced kidney injury is a major clinical problem and causes drug attrition in the pharmaceutical industry. To better predict drug-induced kidney injury, kidney in vitro cultures with enhanced physiologic relevance are developed. To mimic the proximal tubule, the main site of adverse drug reactions in the kidney, human-derived renal proximal tubule epithelial cells (HRPTECs) were injected in one of the channels of dual-channel Nortis chips and perfused for 7 days. Tubes of HRPTECs demonstrated expression of tight junction protein 1 (zona occludens-1), lotus lectin, and primary cilia with localization at the apical membrane, indicating an intact proximal tubule brush border. Gene expression of cisplatin efflux transporters multidrug and toxin extrusion transporter (MATE) 1 (SLC47A1) and MATE2-k (SLC47A2) and megalin endocytosis receptor increased 19.9 ± 5.0–, 23.2 ± 8.4–, and 106 ± 33–fold, respectively, in chip cultures compared with 2-dimensional cultures. Moreover, organic cation transporter 2 (OCT2) (SLC22A2) was localized exclusively on the basolateral membrane. When infused from the basolateral compartment, cisplatin (25 µM, 72 hours) induced toxicity, which was evident as reduced cell number and reduced barrier integrity compared with vehicle-treated chip cultures. Coexposure with the OCT2 inhibitor cimetidine (1 mM) abolished cisplatin toxicity. In contrast, infusion of cisplatin from the apical compartment did not induce toxicity, which was in line with polarized localization of cisplatin uptake transport proteins, including OCT2. In conclusion, we developed a dual channel human kidney proximal tubule-on-a-chip with a polarized epithelium, restricting cisplatin sensitivity to the basolateral membrane and suggesting improved physiologic relevance over single-compartment models. Its implementation in drug discovery holds promise to improve future in vitro drug-induced kidney injury studies.
SIGNIFICANCE STATEMENT Human-derived kidney proximal tubule cells retained characteristics of epithelial polarization in vitro when cultured in the kidney-on-a-chip, and the dual-channel construction allowed for drug exposure using the physiologically relevant compartment. Therefore, cell polarization–dependent cisplatin toxicity could be replicated for the first time in a kidney proximal tubule-on-a-chip. The use of this physiologically relevant model in drug discovery has potential to aid identification of safe novel drugs and contribute to reducing attrition rates due to drug-induced kidney injury.
Footnotes
- Received April 24, 2020.
- Accepted September 23, 2020.
↵1 Current affiliation: CVRM Safety, Clinical Pharmacology and Safety Sciences, AstraZeneca, Gothenburg, Sweden.
↵2 Current affiliation: Department of Pharmaceutics, University of Washington, Seattle, Washington.
↵3 Current affiliation: Kidney Research Institute, University of Washington, Seattle, Washington.
This work was supported in part by the AstraZeneca Postdoc Programme.
Primary laboratory of origin: CVRM Safety, Clinical Pharmacology and Safety Sciences, R&D, AstraZeneca, Gothenburg, Sweden.
T.T.G.N., M.P., and A.-K.S. are employees of AstraZeneca AB, Gothenburg, Sweden. T.T.G.N. is a fellow of the AstraZeneca Postdoc Programme.
This work was previously presented as follows in the cited meeting abstract: Nieskens TTG and Sjögren A-K (2019) A functionally polarized human kidney proximal tubule-on-a-chip for advanced drug-induced nephrotoxicity studies. Winter-meeting in the Danish Society for Pharmacology and Toxicology; 2019 Dec 02; Copenhagen, Denmark.
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- Copyright © 2020 by The American Society for Pharmacology and Experimental Therapeutics
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