1. Novel Cell-Based Assay for Identification of LRRK2 Inhibitors Using Its Aberrant Regulation of a Pluripotency Gene.
- Author
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Ramonet D and Dietz GPH
- Subjects
- Cell Differentiation genetics, Cell Line, Tumor, Gene Expression, Gene Knockdown Techniques, High-Throughput Screening Assays, Humans, Leucine-Rich Repeat Serine-Threonine Protein Kinase-2 genetics, Leucine-Rich Repeat Serine-Threonine Protein Kinase-2 metabolism, Mutation, Nanog Homeobox Protein genetics, Neurites metabolism, Neurons cytology, Neurons metabolism, Promoter Regions, Genetic, Drug Screening Assays, Antitumor methods, Gene Expression Regulation drug effects, Leucine-Rich Repeat Serine-Threonine Protein Kinase-2 antagonists & inhibitors, Protein Kinase Inhibitors pharmacology
- Abstract
Mutations in the gene encoding leucine-rich repeat kinase 2 ( LRRK2 ), such as the G2019S mutation, are the most common cause of familial Parkinson's disease (PD). The G2019S mutation impairs neurite outgrowth. We hypothesized that those effects could be related to an altered expression of pluripotency genes, which may provide a readout for a screening assay based on LRRK2 function. Here, we show that the G2019S mutation mediates a sustained aberrant upregulation of the transcription factors Nanog and Oct4 that in wild-type are downregulated after differentiation. The aberrant regulation of Nanog can be concentration dependently reversed by LRRK2 tool inhibitors. Building on this knowledge, we developed an assay for the identification and assessment of compounds that inhibit the aberrant pathophysiological activity of mutant LRRK2 . Furthermore, the aberrant neural pluripotency is consistent with Parkinson's pathophysiology and with the epidemiological association between the G2019S genotype and cancer risk.
- Published
- 2020
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