Suter-Dick, Laura
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Suter-Dick, Laura
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- PublikationNephrotoxicity and Kidney Transport Assessment on 3D Perfused Proximal Tubules(08/2018) Vormann, Marianne K.; Gijzen, Linda; Hutter, Simon; Boot, Lisette; Nicolas, Arnaud; van den Heuvel, Angelique; Vriend, Jelle; Ng, Chee Ping; Nieskens, Tom T.G.; van Duinen, Vincent; de Wagenaar, Bjorn; Masereeuw, Rosalinde; Suter-Dick, Laura; Trietsch, Sebastian J.; Wilmer, Martijn; Joore, Jos; Vulto, Paul; Lanz, Henriette [in: The AAPS Journal]Proximal tubules in the kidney play a crucial role in reabsorbing and eliminating substrates from the body into the urine, leading to high local concentrations of xenobiotics. This makes the proximal tubule a major target for drug toxicity that needs to be evaluated during the drug development process. Here, we describe an advanced in vitro model consisting of fully polarized renal proximal tubular epithelial cells cultured in a microfluidic system. Up to 40 leak-tight tubules were cultured on this platform that provides access to the basolateral as well as the apical side of the epithelial cells. Exposure to the nephrotoxicant cisplatin caused a dose-dependent disruption of the epithelial barrier, a decrease in viability, an increase in effluent LDH activity, and changes in expression of tight-junction marker zona-occludence 1, actin, and DNA-damage marker H2A.X, as detected by immunostaining. Activity and inhibition of the efflux pumps P-glycoprotein (P-gp) and multidrug resistance protein (MRP) were demonstrated using fluorescence-based transporter assays. In addition, the transepithelial transport function from the basolateral to the apical side of the proximal tubule was studied. The apparent permeability of the fluorescent P-gp substrate rhodamine 123 was decreased by 35% by co-incubation with cyclosporin A. Furthermore, the activity of the glucose transporter SGLT2 was demonstrated using the fluorescent glucose analog 6-NBDG which was sensitive to inhibition by phlorizin. Our results demonstrate that we developed a functional 3D perfused proximal tubule model with advanced renal epithelial characteristics that can be used for drug screening studies.01A - Beitrag in wissenschaftlicher Zeitschrift
- PublikationHigh-Throughput Screening of Drug-Transporter Interactions in a 3D Microfluidic Renal Proximal Tubule on a Chip(07/2018) Vriend, Jelle; Nieskens, Tom T.G.; Vormann, Marianne K.; van den Berge, Batholomeus T.; van den Heuvel, Angelique; Russel, Frans G.M.; Suter-Dick, Laura; Lanz, Henriette; Vulto, Paul; Masereeuw, Rosalinde; Wilmer, Martijn [in: The AAPS Journal]Drug-transporter interactions could impact renal drug clearance and should ideally be detected in early stages of drug development to avoid toxicity-related withdrawals in later stages. This requires reliable and robust assays for which current high-throughput screenings have, however, poor predictability. Kidney-on-a-chip platforms have the potential to improve predictability, but often lack compatibility with high-content detection platforms. Here, we combined conditionally immortalized proximal tubule epithelial cells overexpressing organic anion transporter 1 (ciPTEC-OAT1) with the microfluidic titer plate OrganoPlate to develop a screenings assay for renal drug-transporter interactions. In this platform, apical localization of F-actin and intracellular tight-junction protein zonula occludens-1 (ZO-1) indicated appropriate cell polarization. Gene expression levels of the drug transporters organic anion transporter 1 (OAT1; SLC22A6), organic cation transporter 2 (OCT2; SLC22A2), P-glycoprotein (P-gp; ABCB1), and multidrug resistance-associated protein 2 and 4 (MRP2/4; ABCC2/4) were similar levels to 2D static cultures. Functionality of the efflux transporters P-gp and MRP2/4 was studied as proof-of-concept for 3D assays using calcein-AM and 5-chloromethylfluorescein-diacetate (CMFDA), respectively. Confocal imaging demonstrated a 4.4 ± 0.2-fold increase in calcein accumulation upon P-gp inhibition using PSC833. For MRP2/4, a 3.0 ± 0.2-fold increased accumulation of glutathione-methylfluorescein (GS-MF) was observed upon inhibition with a combination of PSC833, MK571, and KO143. Semi-quantitative image processing methods for P-gp and MRP2/4 was demonstrated with corresponding Z'-factors of 0.1 ± 0.3 and 0.4 ± 0.1, respectively. In conclusion, we demonstrate a 3D microfluidic PTEC model valuable for screening of drug-transporter interactions that further allows multiplexing of endpoint read-outs for drug-transporter interactions and toxicity01A - Beitrag in wissenschaftlicher Zeitschrift
- PublikationP12-040 High throughput in vitro system for nephrotoxicity testing(2015) Suter-Dick, Laura; Prétôt, René; Weston, Anna; Wegner, Irene; Wilmer, Martijn; Nieskens, Tom T.G.; Vulto, Paul; Joore, Jos; Lanz, Henriette; Masereeuw, Rosalinde06 - Präsentation