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Thallium-sensitive fluorescent assay reveals loperamide as a new inhibitor of the potassium channel Kv10.1

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Abstract

Background

Ion channels have been proposed as therapeutic targets for different types of malignancies. One of the most studied ion channels in cancer is the voltage-gated potassium channel ether-à-go-go 1 or Kv10.1. Various studies have shown that Kv10.1 expression induces the proliferation of several cancer cell lines and in vivo tumor models, while blocking or silencing inhibits proliferation. Kv10.1 is a promising target for drug discovery modulators that could be used in cancer treatment. This work aimed to screen for new Kv10.1 channel modulators using a thallium influx-based assay.

Methods

Pharmacological effects of small molecules on Kv10.1 channel activity were studied using a thallium-based fluorescent assay and patch-clamp electrophysiological recordings, both performed in HEK293 stably expressing the human Kv10.1 potassium channel.

Results

In thallium-sensitive fluorescent assays, we found that the small molecules loperamide and amitriptyline exert a potent inhibition on the activity of the oncogenic potassium channel Kv10.1. These results were confirmed by electrophysiological recordings, which showed that loperamide and amitriptyline decreased the amplitude of Kv10.1 currents in a dose-dependent manner. Both drugs could be promising tools for further studies.

Conclusions

Thallium-sensitive fluorescent assay represents a reliable methodological tool for the primary screening of different molecules with potential activity on Kv10.1 channels or other K+ channels.

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Abbreviations

AMI:

Amitriptyline

Eag1:

Human ether-à-go-go potassium channel 1

HEK-WT:

HEK293 wild-type cells

HEK-Kv10.1:

HEK293 cells stably expressing the human Kv10.1 potassium channel

LP:

Loperamide

SR:

SR 33805 oxalate

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Funding

This work was supported by Grants: 314839, 21887 and 315803 from Consejo Nacional de Ciencia y Tecnología (CONACYT) and PAPIIT-UNAM IG200119 to AH-C, and SEP-CONACYT CB2017-2018-A1-S-13646 to E.L. We thank Fis. Cesar Oliver Lara Figueroa and Dr. Luisa Duran Pasten for technical assistance. The English revision by Dr. Bristol Denlinger is also acknowledged.

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Authors and Affiliations

  1. Laboratorio Nacional de Canalopatías, Instituto de Fisiología Celular, Universidad Nacional Autónoma de México, Circuito Exterior s/n, C.U. 04510, Mexico City, Mexico

    Arlet Loza-Huerta, Edgar Milo, Arturo Picones, Arturo Hernández-Cruz & Enoch Luis

  2. Departamento de Neurociencia Cognitiva, Instituto de Fisiología Celular, Universidad Nacional Autónoma de México, Circuito Exterior s/n, C.U. 04510, Mexico City, Mexico

    Arturo Hernández-Cruz

  3. Cátedras CONACYT - Instituto de Fisiología Celular, Universidad Nacional Autónoma de México, Circuito Exterior s/n, C.U. 04510, Mexico City, Mexico

    Enoch Luis

Authors
  1. Arlet Loza-Huerta
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  2. Edgar Milo
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Contributions

AL–H, EM and EL carried out the experiments; EL and AH-C designed, analyzed, and interpreted the results; EL wrote the manuscript. All the authors read and approved the final manuscript.

Corresponding author

Correspondence to Enoch Luis.

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All authors have no conflict of interest.

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Loza-Huerta, A., Milo, E., Picones, A. et al. Thallium-sensitive fluorescent assay reveals loperamide as a new inhibitor of the potassium channel Kv10.1. Pharmacol. Rep 73, 1744–1753 (2021). https://doi.org/10.1007/s43440-021-00304-5

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  • DOI: https://doi.org/10.1007/s43440-021-00304-5

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