title: Fluorescent Parkin Cell-Based Assay Development for the Screening of Drugs against Parkinson Disease.
abstract: Parkinson disease (PD) is a prevalent neurodegenerative disease characterized by selective degeneration of dopaminergic neurons in the substantia nigra, causing tremor and motor impairment. Parkin protein, whose mutants are the cause of Parkinson disease type 2 (PARK2), has been mechanistically linked to the regulation of apoptosis and the turnover of damaged mitochondria. Several studies have implicated aberrant mitochondria as a key contributor to the development of PD. In the attempt to discover new drugs, high-content cell-based assays are becoming more important to mimic the nature of biological processes and their diversifications in diseases and will be essential for lead identification and the optimization of therapeutic candidates. We have developed a novel fluorescence cell-based assay for high-content screening to find compounds that can promote the mitochondrial localization of Parkin without severe mitochondrial damage induction. In this work, this model was used to screen a library of 1280 compounds. After the screening campaign, the positive compounds were chosen for further testing, based on the strength of the initial response and lack of cytotoxicity. These results indicated that this Parkin cell-based assay is a robust (Z' > 0.5) and valid strategy to test potential candidates for preclinical studies.
authors.affiliation: 1 Innoprot (Innovative Technologies in Biological Systems), Derio, Spain.
authors.forename: Patricia
authors.initials: P
authors.lastname: Villacé
authors.affiliation: 1 Innoprot (Innovative Technologies in Biological Systems), Derio, Spain.
authors.forename: Rosa M
authors.initials: RM
authors.lastname: Mella
authors.affiliation: 1 Innoprot (Innovative Technologies in Biological Systems), Derio, Spain.
authors.forename: Meritxell
authors.initials: M
authors.lastname: Roura-Ferrer
authors.affiliation: 1 Innoprot (Innovative Technologies in Biological Systems), Derio, Spain.
authors.forename: María
authors.initials: M
authors.lastname: Valcárcel
authors.affiliation: 1 Innoprot (Innovative Technologies in Biological Systems), Derio, Spain.
authors.forename: Clarisa
authors.initials: C
authors.lastname: Salado
authors.affiliation: 1 Innoprot (Innovative Technologies in Biological Systems), Derio, Spain.
authors.forename: Amaia
authors.initials: A
authors.lastname: Castilla
authors.affiliation: 1 Innoprot (Innovative Technologies in Biological Systems), Derio, Spain.
authors.forename: Danel
authors.initials: D
authors.lastname: Kortazar
mesh_terms: D001681:Biological Assay / Q000379:methods*; D045744:Cell Line, Tumor; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D005453:Fluorescence; D006801:Humans; D008928:Mitochondria / Q000378:metabolism; D010300:Parkinson Disease / Q000188:drug therapy* / Q000378:metabolism; D011442:Proscillaridin / Q000627:therapeutic use; D012235:Rhodamines; D044767:Ubiquitin-Protein Ligases / Q000378:metabolism*
chemical_list: D012235:Rhodamines; C401833:tetramethylrhodamine methyl ester; D044767:Ubiquitin-Protein Ligases; C111567:parkin protein; D011442:Proscillaridin
keywords: Parkinson’s disease; cell-based assay; high-content screening; mitochondria