High-Throughput Screening of Drug-Transporter Interactions in a 3D Microfluidic Renal Proximal Tubule on a Chip
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Autor:innen
Vriend, Jelle
Nieskens, Tom T.G.
Vormann, Marianne K.
van den Berge, Batholomeus T.
van den Heuvel, Angelique
Russel, Frans G.M.
Lanz, Henriette
Vulto, Paul
Masereeuw, Rosalinde
Autor:in (Körperschaft)
Publikationsdatum
07/2018
Typ der Arbeit
Studiengang
Typ
01 - Zeitschriftenartikel, Journalartikel oder Magazin
Herausgeber:innen
Herausgeber:in (Körperschaft)
Betreuer:in
Übergeordnetes Werk
The AAPS Journal
Themenheft
Reihe / Serie
Jahrgang / Band
20
Ausgabe / Nummer
5
Seiten / Dauer
Patentnummer
Verlag / Herausgebende Institution
Verlagsort / Veranstaltungsort
Auflage
Version
Programmiersprache
Abtretungsempfänger:in
Praxispartner:in/Auftraggeber:in
Zusammenfassung
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 toxicity
Schlagwörter
drug screening, drug-transporter interaction, Efflux transport, microfluidics, nephrotoxiciy
Fachgebiet (DDC)
Veranstaltung
Startdatum der Ausstellung
Enddatum der Ausstellung
Startdatum der Konferenz
Enddatum der Konferenz
Datum der letzten Prüfung
ISBN
ISSN
Sprache
Englisch
Während FHNW Zugehörigkeit erstellt
Ja
Publikationsstatus
Veröffentlicht
Begutachtung
Peer-Review der ganzen Publikation
Open Access-Status
Lizenz
Zitation
VRIEND, Jelle, Tom T.G. NIESKENS, Marianne K. VORMANN, Batholomeus T. VAN DEN BERGE, Angelique VAN DEN HEUVEL, Frans G.M. RUSSEL, Laura SUTER-DICK, Henriette LANZ, Paul VULTO, Rosalinde MASEREEUW und Martijn WILMER, 2018. High-Throughput Screening of Drug-Transporter Interactions in a 3D Microfluidic Renal Proximal Tubule on a Chip. The AAPS Journal. Juli 2018. Bd. 20, Nr. 5. DOI 10.1208/s12248-018-0247-0. Verfügbar unter: http://hdl.handle.net/11654/26933