A novel yeast-based screening system for potential compounds that can alleviate human α-synuclein toxicity

Area of Science:

• Neuroscience
• Biochemistry
• Pharmacology

Background:

• Alpha-synuclein (α-syn) aggregation is a key pathological feature of Parkinson's disease (PD).
• Developing effective screening methods to identify compounds that mitigate α-syn toxicity is crucial for PD drug discovery.
• Existing screening methods may lack the sensitivity or throughput required for comprehensive drug discovery.

Purpose of the Study:

• To establish a sensitive and specific yeast-based screening system for compounds targeting α-syn toxicity.
• To identify compounds that either inhibit α-syn aggregation or promote its degradation.
• To validate the system's efficacy using a known α-syn aggregation inhibitor.

Main Methods:

• Development of a hypersensitive yeast strain (rsp5^A401E-mutant) for α-syn aggregation screening.
• Overexpression of GFP-fused α-syn in the engineered yeast strain.
• Quantification of α-syn toxicity alleviation using resazurin fluorescence assay.

Main Results:

• The established yeast assay system demonstrated high sensitivity in detecting α-syn toxicity alleviation.
• Baicalein, a known α-syn aggregation inhibitor, showed enhanced efficacy in the engineered yeast strain compared to the wild type.
• The system detected baicalein at concentrations as low as 3.13 µmol L^-1, surpassing the sensitivity of previous in vitro assays.

Conclusions:

• The developed yeast-based system is effective for screening compounds that alleviate α-syn toxicity with high sensitivity and specificity.
• This system can accelerate the discovery of novel neuroprotective drug candidates for Parkinson's disease.
• The assay facilitates the identification of compounds that modulate α-syn aggregation or enhance its degradation.