MPP<sup>+</sup>-Induced Changes in Cellular Impedance as a Measure for Organic Cation Transporter (SLC22A1-3) Activity and Inhibition
The organic cation transporters OCT1-3 (<i>SLC22A1-3</i>) 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, scre...
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| Format: | Article |
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MDPI AG
2022-01-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/23/3/1203 |
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| author | Tamara A. M. Mocking Hubert J. Sijben Yimé W. Vermeulen Adriaan P. IJzerman Laura H. Heitman |
| author_facet | Tamara A. M. Mocking Hubert J. Sijben Yimé W. Vermeulen Adriaan P. IJzerman Laura H. Heitman |
| author_sort | Tamara A. M. Mocking |
| collection | DOAJ |
| description | The organic cation transporters OCT1-3 (<i>SLC22A1-3</i>) 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<sup>+</sup>. Uptake of MPP<sup>+</sup> by OCTs induced concentration-dependent changes in cellular impedance that were inhibited by decynium-22, corticosterone, and Tyrosine Kinase inhibitors. OCT-mediated MPP<sup>+</sup> 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<sup>+</sup> transporting solute carrier proteins (SLCs) in general. |
| first_indexed | 2024-03-09T23:49:46Z |
| format | Article |
| id | doaj.art-7f276ced9d5640cd90f941732da39cd9 |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-09T23:49:46Z |
| publishDate | 2022-01-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-7f276ced9d5640cd90f941732da39cd92023-11-23T16:36:39ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672022-01-01233120310.3390/ijms23031203MPP<sup>+</sup>-Induced Changes in Cellular Impedance as a Measure for Organic Cation Transporter (SLC22A1-3) Activity and InhibitionTamara A. M. Mocking0Hubert J. Sijben1Yimé W. Vermeulen2Adriaan P. IJzerman3Laura H. Heitman4Division of Drug Discovery and Safety, Leiden Academic Centre for Drug Research, Leiden University, Einsteinweg 55, 2333 CC Leiden, The NetherlandsDivision of Drug Discovery and Safety, Leiden Academic Centre for Drug Research, Leiden University, Einsteinweg 55, 2333 CC Leiden, The NetherlandsDivision of Drug Discovery and Safety, Leiden Academic Centre for Drug Research, Leiden University, Einsteinweg 55, 2333 CC Leiden, The NetherlandsDivision of Drug Discovery and Safety, Leiden Academic Centre for Drug Research, Leiden University, Einsteinweg 55, 2333 CC Leiden, The NetherlandsDivision of Drug Discovery and Safety, Leiden Academic Centre for Drug Research, Leiden University, Einsteinweg 55, 2333 CC Leiden, The NetherlandsThe organic cation transporters OCT1-3 (<i>SLC22A1-3</i>) 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<sup>+</sup>. Uptake of MPP<sup>+</sup> by OCTs induced concentration-dependent changes in cellular impedance that were inhibited by decynium-22, corticosterone, and Tyrosine Kinase inhibitors. OCT-mediated MPP<sup>+</sup> 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<sup>+</sup> transporting solute carrier proteins (SLCs) in general.https://www.mdpi.com/1422-0067/23/3/1203OCTlabel-freexCELLigenceMPP<sup>+</sup>transportSLC22A1-3 |
| spellingShingle | Tamara A. M. Mocking Hubert J. Sijben Yimé W. Vermeulen Adriaan P. IJzerman Laura H. Heitman MPP<sup>+</sup>-Induced Changes in Cellular Impedance as a Measure for Organic Cation Transporter (SLC22A1-3) Activity and Inhibition International Journal of Molecular Sciences OCT label-free xCELLigence MPP<sup>+</sup> transport SLC22A1-3 |
| title | MPP<sup>+</sup>-Induced Changes in Cellular Impedance as a Measure for Organic Cation Transporter (SLC22A1-3) Activity and Inhibition |
| title_full | MPP<sup>+</sup>-Induced Changes in Cellular Impedance as a Measure for Organic Cation Transporter (SLC22A1-3) Activity and Inhibition |
| title_fullStr | MPP<sup>+</sup>-Induced Changes in Cellular Impedance as a Measure for Organic Cation Transporter (SLC22A1-3) Activity and Inhibition |
| title_full_unstemmed | MPP<sup>+</sup>-Induced Changes in Cellular Impedance as a Measure for Organic Cation Transporter (SLC22A1-3) Activity and Inhibition |
| title_short | MPP<sup>+</sup>-Induced Changes in Cellular Impedance as a Measure for Organic Cation Transporter (SLC22A1-3) Activity and Inhibition |
| title_sort | mpp sup sup induced changes in cellular impedance as a measure for organic cation transporter slc22a1 3 activity and inhibition |
| topic | OCT label-free xCELLigence MPP<sup>+</sup> transport SLC22A1-3 |
| url | https://www.mdpi.com/1422-0067/23/3/1203 |
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