KMID : 0606920230310010127
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Biomolecules & Therapeutics 2023 Volume.31 No. 1 p.127 ~ p.138
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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
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Lin Chih-Hsin
Hsieh Yu-Shao Sun Ying-Chieh Huang Wun-Han Chen Shu-Ling Weng Zheng-Kui Lin Te-Hsien Wu Yih-Ru Chang Kuo-Hsuan Huang Hei-Jen Lee Guan-Chiun Hsieh-Li Hsiu Mei Lee-Chen Guey-Jen
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Abstract
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Glycogen synthase kinase-3¥â (GSK-3¥â) 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-3¥â 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 ¥ÄK280. In the kinase assay for these 7 compounds, residual GSK-3¥â 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 ¥ÄK280 TauRD-DsRed folding reporter. In SH-SY5Y cells expressing ¥ÄK280 TauRD, neither VB-030 nor VB-037 increased expression of GSK-3¥á Ser21 or GSK-3¥â Ser9. Among extracellular signal-regulated kinase (ERK), AKT serine/threonine kinase 1 (AKT), mitogen-activated protein kinase 14 (P38) and mitogen-activated 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 ¥ÄK280 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-3¥â kinase activity and/or p-P38 (Thr180/Tyr182), both compounds may serve as promising candidates to reduce Tau aggregation/cytotoxicity for AD treatment.
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KEYWORD
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GSK-3¥â kinase inhibitor, Alzheimer¡¯s disease, Virtual screening, Enzyme assay, Cell assay, Mouse hippocampal primary culture
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