Abstract
Objective: Reducing the burden of α-synuclein oligomeric species represents a promising approach for disease-modifying therapies against synucleinopathies such as Parkinson's disease and dementia with Lewy bodies. However, the lack of efficient drug discovery strategies that specifically target α-synuclein oligomers has been a limitation to drug discovery programs.Research design and methods: Here we describe an innovative strategy that harnesses the power of bimolecular protein-fragment complementation to monitor synuclein-synuclein interactions. We have developed two robust models to monitor α-synuclein oligomerization by generating novel stable cell lines expressing α-synuclein fusion proteins for either fluorescent or bioluminescent protein-fragment complementation under the tetracycline-controlled transcriptional activation system.Main outcome measures: A pilot screen was performed resulting in the identification of two potential hits, a p38 MAPK inhibitor and a casein kinase 2 inhibitor, thereby demonstrating the suitability of our protein-fragment complementation assay for the measurement of α-synuclein oligomerization in living cells at high throughput.Conclusions: The application of the strategy described herein to monitor α-synuclein oligomer formation in living cells with high throughput will facilitate drug discovery efforts for disease-modifying therapies against synucleinopathies and other proteinopathies.
Original language | English (US) |
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Pages (from-to) | 589-603 |
Number of pages | 15 |
Journal | Expert opinion on therapeutic targets |
Volume | 19 |
Issue number | 5 |
DOIs | |
State | Published - May 1 2015 |
Keywords
- High-throughput screening
- Oligomers
- Parkinson's disease
- Protein-fragment complementation
- Synucleinopathy
- α-synuclein
ASJC Scopus subject areas
- Molecular Medicine
- Pharmacology
- Drug Discovery
- Clinical Biochemistry