High-throughput screening against protein:protein interaction interfaces reveals anti-cancer therapeutics as potent modulators of the voltage-gated Na+ channel complex

Paul A. Wadsworth, Oluwarotimi Folorunso, Nghi Nguyen, Aditya K. Singh, Daniela D’Amico, Reid T. Powell, David Brunell, John Allen, Clifford Stephan, Fernanda Laezza

Research output: Contribution to journalArticle

Abstract

Multiple voltage-gated Na+ (Nav) channelopathies can be ascribed to subtle changes in the Nav macromolecular complex. Fibroblast growth factor 14 (FGF14) is a functionally relevant component of the Nav1.6 channel complex, a causative link to spinocerebellar ataxia 27 (SCA27) and an emerging risk factor for neuropsychiatric disorders. Yet, how this protein:channel complex is regulated in the cell is still poorly understood. To search for key cellular pathways upstream of the FGF14:Nav1.6 complex, we have developed, miniaturized and optimized an in-cell assay in 384-well plates by stably reconstituting the FGF14:Nav1.6 complex using the split-luciferase complementation assay. We then conducted a high-throughput screening (HTS) of 267 FDA-approved compounds targeting known mediators of cellular signaling. Of the 65 hits initially detected, 24 were excluded based on counter-screening and cellular toxicity. Based on target analysis, potency and dose-response relationships, 5 compounds were subsequently repurchased for validation and confirmed as hits. Among those, the tyrosine kinase inhibitor lestaurtinib was highest ranked, exhibiting submicromolar inhibition of FGF14:Nav1.6 assembly. While providing evidence for a robust in-cell HTS platform that can be adapted to search for any channelopathy-associated regulatory proteins, these results lay the potential groundwork for repurposing cancer drugs for neuropsychopharmacology.

Original languageEnglish (US)
Article number16890
JournalScientific reports
Volume9
Issue number1
DOIs
StatePublished - Dec 1 2019

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Channelopathies
Neoplasms
Proteins
Drug Repositioning
Macromolecular Substances
Therapeutics
Luciferases
Protein-Tyrosine Kinases
fibroblast growth factor 14
lestaurtinib
Spinocerebellar ataxia 27

ASJC Scopus subject areas

  • General

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High-throughput screening against protein:protein interaction interfaces reveals anti-cancer therapeutics as potent modulators of the voltage-gated Na+ channel complex. / Wadsworth, Paul A.; Folorunso, Oluwarotimi; Nguyen, Nghi; Singh, Aditya K.; D’Amico, Daniela; Powell, Reid T.; Brunell, David; Allen, John; Stephan, Clifford; Laezza, Fernanda.

In: Scientific reports, Vol. 9, No. 1, 16890, 01.12.2019.

Research output: Contribution to journalArticle

Wadsworth, Paul A. ; Folorunso, Oluwarotimi ; Nguyen, Nghi ; Singh, Aditya K. ; D’Amico, Daniela ; Powell, Reid T. ; Brunell, David ; Allen, John ; Stephan, Clifford ; Laezza, Fernanda. / High-throughput screening against protein:protein interaction interfaces reveals anti-cancer therapeutics as potent modulators of the voltage-gated Na+ channel complex. In: Scientific reports. 2019 ; Vol. 9, No. 1.
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