Biomolecules & Therapeutics

The Korean Society of Applied Pharmacology (한국응용약물학회)
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Virtual Screening and Testing of GSK-3 Inhibitors Using Human SH-SY5Y Cells Expressing Tau Folding Reporter and Mouse Hippocampal Primary Culture under Tau Cytotoxicity

  • Chih-Hsin, Lin (School of Life Science, National Taiwan Normal University) ;
  • Yu-Shao, Hsieh (Department of Chemistry, National Taiwan Normal University) ;
  • Ying-Chieh, Sun (Department of Chemistry, National Taiwan Normal University) ;
  • Wun-Han, Huang (School of Life Science, National Taiwan Normal University) ;
  • Shu-Ling, Chen (School of Life Science, National Taiwan Normal University) ;
  • Zheng-Kui, Weng (School of Life Science, National Taiwan Normal University) ;
  • Te-Hsien, Lin (School of Life Science, National Taiwan Normal University) ;
  • Yih-Ru, Wu (Department of Neurology, Chang Gung Memorial Hospital) ;
  • Kuo-Hsuan, Chang (Department of Neurology, Chang Gung Memorial Hospital) ;
  • Hei-Jen, Huang (Department of Nursing, Mackay Junior College of Medicine, Nursing and Management) ;
  • Guan-Chiun, Lee (School of Life Science, National Taiwan Normal University) ;
  • Hsiu Mei, Hsieh-Li (School of Life Science, National Taiwan Normal University) ;
  • Guey-Jen, Lee-Chen (School of Life Science, National Taiwan Normal University)
  • Received : 2022.03.11
  • Accepted : 2022.05.24
  • Published : 2023.01.01
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Abstract

Glycogen synthase kinase-3β (GSK-3β) is an important serine/threonine kinase that implicates in multiple cellular processes and links with the neurodegenerative diseases including Alzheimer's disease (AD). In this study, structure-based virtual screening was performed to search database for compounds targeting GSK-3β from Enamine's screening collection. Of the top-ranked compounds, 7 primary hits underwent a luminescent kinase assay and a cell assay using human neuroblastoma SH-SY5Y cells expressing Tau repeat domain (TauRD) with pro-aggregant mutation ΔK280. In the kinase assay for these 7 compounds, residual GSK-3β activities ranged from 36.1% to 90.0% were detected at the IC50 of SB-216763. In the cell assay, only compounds VB-030 and VB-037 reduced Tau aggregation in SH-SY5Y cells expressing ΔK280 TauRD-DsRed folding reporter. In SH-SY5Y cells expressing ΔK280 TauRD, neither VB-030 nor VB-037 increased expression of GSK-3α Ser21 or GSK-3β Ser9. Among extracellular signal-regulated kinase (ERK), AKT serine/threonine kinase 1 (AKT), mitogen-activated protein kinase 14 (P38) and mitogenactivated protein kinase 8 (JNK) which modulate Tau phosphorylation, VB-037 attenuated active phosphorylation of P38 Thr180/ Tyr182, whereas VB-030 had no effect on the phosphorylation status of ERK, AKT, P38 or JNK. However, both VB-030 and VB-037 reduced endogenous Tau phosphorylation at Ser202, Thr231, Ser396 and Ser404 in neuronally differentiated SH-SY5Y expressing ΔK280 TauRD. In addition, VB-030 and VB-037 further improved neuronal survival and/or neurite length and branch in mouse hippocampal primary culture under Tau cytotoxicity. Overall, through inhibiting GSK-3β kinase activity and/or p-P38 (Thr180/Tyr182), both compounds may serve as promising candidates to reduce Tau aggregation/cytotoxicity for AD treatment.

Keywords

Acknowledgement

We thank the Molecular Imaging Core Facility of National Taiwan Normal University for the technical assistance. This work was supported by the grants 103-2321-B-182-008, 103-2321-B-003-003, 104-2325-B-003-001 and 104-2325- B-003-003 from the Ministry of Science and Technology, and 104T3040B05 and 104T3040B07 from National Taiwan Normal University, Taipei, Taiwan.

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