1. MPP + -Induced Changes in Cellular Impedance as a Measure for Organic Cation Transporter (SLC22A1-3) Activity and Inhibition.
- Author
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Mocking TAM, Sijben HJ, Vermeulen YW, IJzerman AP, and Heitman LH
- Subjects
- 1-Methyl-4-phenylpyridinium adverse effects, Biological Transport, Biological Transport, Active, HEK293 Cells, Herbicides adverse effects, Humans, Organic Cation Transport Proteins antagonists & inhibitors, Organic Cation Transport Proteins genetics, Organic Cation Transporter 1 antagonists & inhibitors, Organic Cation Transporter 1 genetics, Organic Cation Transporter 2 antagonists & inhibitors, Organic Cation Transporter 2 genetics, Electric Impedance, Gene Expression Regulation drug effects, Organic Cation Transport Proteins metabolism, Organic Cation Transporter 1 metabolism, Organic Cation Transporter 2 metabolism
- Abstract
The organic cation transporters OCT1-3 ( SLC22A1-3 ) facilitate the transport of cationic endo- and xenobiotics and are important mediators of drug distribution and elimination. Their polyspecific nature makes OCTs highly susceptible to drug-drug interactions (DDIs). Currently, screening of OCT inhibitors depends on uptake assays that require labeled substrates to detect transport activity. However, these uptake assays have several limitations. Hence, there is a need to develop novel assays to study OCT activity in a physiological relevant environment without the need to label the substrate. Here, a label-free impedance-based transport assay is established that detects OCT-mediated transport activity and inhibition utilizing the neurotoxin MPP
+ . Uptake of MPP+ by OCTs induced concentration-dependent changes in cellular impedance that were inhibited by decynium-22, corticosterone, and Tyrosine Kinase inhibitors. OCT-mediated MPP+ transport activity and inhibition were quantified on both OCT1-3 overexpressing cells and HeLa cells endogenously expressing OCT3. Moreover, the method presented here is a valuable tool to identify novel inhibitors and potential DDI partners for MPP+ transporting solute carrier proteins (SLCs) in general.- Published
- 2022
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