Cell-Based Screen Using Amyloid Mimic β23 Expression Identifies Peucedanocoumarin III as a Novel Inhibitor of α-Synuclein and Huntingtin Aggregates

• Published in: Molecules and cells 2019, 42(6), , pp.480-494
• Main Authors: Ham, Sangwoo, Kim, Hyojung, Hwang, Seojin, Kang, Hyunook, Yun, Seung Pil, Kim, Sangjune, Kim, Donghoon, Kwon, Hyun Sook, Lee, Yun-Song, Cho, MyoungLae, Shin, Heung-Mook, Choi, Heejung, Chung, Ka Young, Ko, Han Seok, Lee, Gum Hwa, Lee, Yunjong
• Format: Article
• Language: English
• Subjects: alpha-Synuclein - chemistry; Amyloid - chemistry; Coumarins - pharmacology; HEK293 Cells; High-Throughput Screening Assays; Humans; Huntingtin Protein - chemistry; Neuroblastoma; Nuclear Proteins - antagonists & inhibitors; Nuclear Proteins - metabolism; Oxidopamine - pharmacology; Peptides - metabolism; Protein Aggregates - drug effects; Tetracycline - metabolism; Tetracycline - pharmacology; 생물학
• ISSN: 1016-8478; 0219-1032
• DOI: 10.14348/MOLCELLS.2019.0091

Aggregates of disease-causing proteins dysregulate cellular functions, thereby causing neuronal cell loss in diverse neurodegenerative diseases. Although many in vitro or in vivo studies of protein aggregate inhibitors have been performed, a therapeutic strategy to control aggregate toxicity has not been earnestly pursued, partly due to the limitations of available aggregate models. In this study, we established a tetracycline (Tet)-inducible nuclear aggregate (β23) expression model to screen potential lead compounds inhibiting β23-induced toxicity. Highthroughput screening identified several natural compounds as nuclear β23 inhibitors, including peucedanocoumarin III (PCIII). Interestingly, PCIII accelerates disaggregation and proteasomal clearance of both nuclear and cytosolic β23 aggregates and protects SH-SY5Y cells from toxicity induced by β23 expression. Of translational relevance, PCIII disassembled fibrils and enhanced clearance of cytosolic and nuclear protein aggregates in cellular models of huntingtin and α-synuclein aggregation. Moreover, cellular toxicity was diminished with PCIII treatment for polyglutamine (PolyQ)-huntingtin expression and α-synuclein expression in conjunction with 6-hydroxydopamine (6-OHDA) treatment. Importantly, PCIII not only inhibited α-synuclein aggregation but also disaggregated preformed α-synuclein fibrils in vitro . Taken together, our results suggest that a Tet-Off β23 cell model could serve as a robust platform for screening effective lead compounds inhibiting nuclear or cytosolic protein aggregates. Brain-permeable PCIII or its derivatives could be beneficial for eliminating established protein aggregates.