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. 2022 Nov 2;13:971065. doi: 10.3389/fphar.2022.971065

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Copyright © 2022 Buvall, Menzies, Williams, Woollard, Kumar, Granqvist, Fritsch, Feliers, Reznichenko, Gianni, Petrovski, Bendtsen, Bohlooly-Y, Haefliger, Danielson and Hansen.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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In vitro target validation process. In vitro target validation in cultured cells is used as a first approach to validate and screen several targets, aiming to triage targets with strong support for further target validation in advanced in vitro systems. Data from all systems feed into each other to select the most translatable model and define the correct stressor that regulates target and pathway. An in vitro target validation toolbox may comprise various assays, stressors, and readouts, which are chosen based on the disease biology of the target. ADMA, asymmetric dimethylarginine (arginine metabolite); CKD, chronic kidney disease; ECAR, extracellular acidification rate; HGEC, human glomerular endothelial cell; HMOX, hem oxygenase; IHC, immunohistochemistry; iPSC, induced pluripotent stem cell; MMPs, matrix metalloproteinases; MSC, mesenchymal stem cell; OCR, oxygen consumption rate; PAN, pyromycin aminonucleoside; PTEC, proximal tubular epithelial cell; RPTEC, renal proximal tubular epithelial cell; SNP, single nucleotide polymorphism; TGF, transforming growth factor; TLR, toll-like receptor; ROS, reactive oxygen species. 1. Faivre A et al. Front Med (Lausanne) 2021; 8:742072; 2. Imasawa T et al. The International Journal of Biochemistry and Cell Biology 2013; 45:2109–2118.3. Oates JC et al. American Journal of Physiology-Renal Physiology 2022; 322:F309-F321.4. Tang SCW et al. Nature Reviews Nephrology 2020; 16:206–222.5. Lee HW et al. Journal of the American Society of Nephrology 2015; 26:2741–2752.6. Perico L et al. Nature Reviews Nephrology 2016; 12:692–710.7. Prozialeck WC et al. Pharmacology and Therapeutics 2007; 114:74–93.8. Slyne J et al. Nephrology Dialysis Transplantation 2015; 30:iv60-iv67.9. Wieser M et al. American Journal of Physiology-Renal Physiology 2008; 295:F1365-F1375.10. Jourde-Chiche N et al. Nature Reviews Nephrology 2019; 15:87–108.11. Sol M et al. Front Pharmacol 2020; 11:573557.12. Liu Y. Kidney International 2006; 69:213–217.13. Yun CW et al. International Journal of Molecular Sciences 2019; 20:1619.