title: Targeting α-synuclein oligomers by protein-fragment complementation for drug discovery in synucleinopathies.
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.
  
  
  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.
  
  
  METHODS
  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.
authors.affiliation: Mayo Clinic Florida, Neuroscience , 4500 San Pablo road, Jacksonville, 32224, FL , USA McLean.Pamela@mayo.edu.
authors.forename: Simon
authors.initials: S
authors.lastname: Moussaud
authors.forename: Siobhan
authors.initials: S
authors.lastname: Malany
authors.forename: Alka
authors.initials: A
authors.lastname: Mehta
authors.forename: Stefan
authors.initials: S
authors.lastname: Vasile
authors.forename: Layton H
authors.initials: LH
authors.lastname: Smith
authors.forename: Pamela J
authors.initials: PJ
authors.lastname: McLean
mesh_terms: D047390:Casein Kinase II / Q000037:antagonists & inhibitors; D002460:Cell Line; D015195:Drug Design; D055808:Drug Discovery / Q000379:methods*; D057166:High-Throughput Screening Assays; D006801:Humans; D020961:Lewy Body Disease / Q000188:drug therapy* / Q000503:physiopathology; D008954:Models, Biological; D058990:Molecular Targeted Therapy; D010300:Parkinson Disease / Q000188:drug therapy* / Q000503:physiopathology; D010865:Pilot Projects; D055503:Protein Multimerization; D051844:alpha-Synuclein / Q000378:metabolism*; D048051:p38 Mitogen-Activated Protein Kinases / Q000037:antagonists & inhibitors
chemical_list: D051844:alpha-Synuclein; D047390:Casein Kinase II; D048051:p38 Mitogen-Activated Protein Kinases
keywords: Parkinson’s disease; high-throughput screening; oligomers; protein-fragment complementation; synucleinopathy; α-synuclein