Abstract
Multiplications, mutations and dysregulation of the alpha synuclein gene (SNCA) are associated with the demise of dopaminergic neurons and are considered to play important roles in the pathogenesis of familial and sporadic forms of Parkinson’s disease. Regulation of SNCA expression might thus be an appropriate target for treatment. We aimed to identify specific modulators of SNCA transcription, generated CRISPR/Cas9 modified SNCA-GFP-luciferase (LUC) genomic fusion- and control cell lines and screened a library of 1649 bioactive compounds, including the FDA approved drugs. We found no inhibitors but three selective activators which increased SNCA mRNA and protein levels.
Highlights
Multiplications, mutations and dysregulation of the alpha synuclein gene (SNCA) are associated with the demise of dopaminergic neurons and are considered to play important roles in the pathogenesis of familial and sporadic forms of Parkinson’s disease
We performed a LUC reporter-based high throughput screen (HTS) and subsequent room temperature (RT)-qPCR assays to screen a library of 1649 bioactive compounds for transcriptional modifiers of SNCA expression
Among the 25 potential inhibitors we found none which reduced SNCA mRNA levels, but identified three compounds which increased SNCA mRNA and α-syn protein levels (Figs. 3, 4)
Summary
Multiplications, mutations and dysregulation of the alpha synuclein gene (SNCA) are associated with the demise of dopaminergic neurons and are considered to play important roles in the pathogenesis of familial and sporadic forms of Parkinson’s disease. Several modifier screens (genetically or compound modifiers) for α-syn induced toxicity based on α-syn overexpression-models have been performed in different organisms like yeast, E. coli, C. elegans, in rodent- and human cell lines. Mittal and c olleagues[3] performed the first study addressing endogenous SNCA mRNA expression by screening a library of FDA approved compounds and found that β2 adrenoreceptor (β2AR) agonists reduced SNCA mRNA and α-syn protein levels. We had chosen an alternative approach to identify modifiers of SNCA expression and designed a luciferase (LUC) reporter-based high throughput screening of 1649 bioactive drugs including 845 FDA approved compounds in CRISPR/Cas[9] modified human SH-SY5Y neuroblastoma cell lines. We identified three selective activators of SNCA mRNA and α-syn protein levels
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
