Scientific reports, 2019-11, Vol.9 (1), p.16890-15, Article 16890
2019
Volltextzugriff (PDF)
Details
- Autor(en) / Beteiligte
- Titel
- High-throughput screening against protein:protein interaction interfaces reveals anti-cancer therapeutics as potent modulators of the voltage-gated Na + channel complex
- Ist Teil von
- Scientific reports, 2019-11, Vol.9 (1), p.16890-15, Article 16890
- Ort / Verlag
- England: Nature Publishing Group
- Erscheinungsjahr
- 2019
- Quelle
- Free E-Journal (出版社公開部分のみ）
- Beschreibungen/Notizen
- 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.
- Sprache
- Englisch
- Identifikatoren
- ISSN: 2045-2322eISSN: 2045-2322DOI: 10.1038/s41598-019-53110-8Titel-ID: cdi_pubmedcentral_primary_oai_pubmedcentral_nih_gov_6858373
- Format
- –
- Schlagworte
- Antineoplastic Agents - isolation & purification, Antineoplastic Agents - pharmacology, Channelopathy, Dose-response effects, Drug Evaluation, Preclinical - methods, Enzyme inhibitors, Fibroblast growth factors, Fibroblast Growth Factors - agonists, Fibroblast Growth Factors - antagonists & inhibitors, Fibroblast Growth Factors - chemistry, HEK293 Cells, High-throughput screening, High-Throughput Screening Assays - methods, Humans, Interfaces, Macromolecules, Mental disorders, Multiprotein Complexes - agonists, Multiprotein Complexes - antagonists & inhibitors, Multiprotein Complexes - chemistry, NAV1.6 Voltage-Gated Sodium Channel - drug effects, NAV1.6 Voltage-Gated Sodium Channel - metabolism, Neuromodulation, Protein Binding, Protein Interaction Maps - physiology, Protein-tyrosine kinase, Proteins, Regulatory proteins, Risk factors, Sodium channels (voltage-gated), Toxicity, Voltage-Gated Sodium Channel Agonists - isolation & purification, Voltage-Gated Sodium Channel Agonists - pharmacology, Voltage-Gated Sodium Channel Blockers - isolation & purification, Voltage-Gated Sodium Channel Blockers - pharmacology, Voltage-Gated Sodium Channels - drug effects, Voltage-Gated Sodium Channels - metabolism