Mutant Alpha-Synuclein A56P Enhances Toxicity Signal for Drug Screening
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Solution Overview
Problem
Current technologies lack effective tools to identify substances that can prevent or reduce the toxicity of alpha-synuclein, a protein implicated in neurodegenerative diseases such as Parkinson's, which is crucial for understanding and treating synucleinopathies.
Innovation Solution
A mutant human alpha-synuclein with increased toxicity, featuring an amino acid substitution at position 56 from alanine to proline (A56P), along with optional substitutions at positions 76, 127, and 118, is developed. This mutant alpha-synuclein is used in various models to study its toxicity and as a tool for screening potential therapeutic compounds.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If wildtype alpha-synuclein is used in toxicity studies, then the model reflects natural conditions, but the toxicity signal is insufficient for effective drug screening
Solution Approach 1:
The patent applies parameter changes by modifying the amino acid sequence of alpha-synuclein (specifically A56P substitution) to alter its toxicity parameters. This creates a mutant form with enhanced toxic properties that provides a stronger signal for drug screening while maintaining relevance to synucleinopathies.
Solution Approach 2:
Instead of using the natural wildtype form and hoping for sufficient toxicity, the patent inverts the approach by deliberately creating a more toxic mutant form. This inversion allows researchers to work with an enhanced toxicity model that is actually more useful for screening, turning the limitation of weak wildtype toxicity into an advantage.
2Manufacturing precision
If mutant alpha-synuclein with increased aggregation propensity is used, then aggregation studies are enhanced, but the role of different aggregated species in neurotoxicity remains unclear
Solution Approach 1:
The patent applies local quality by making a specific localized substitution (A56P) rather than global modifications. This targeted change at position 56 selectively enhances toxicity while allowing researchers to study specific aspects of aggregation and neurotoxicity relationships in controlled conditions.
3Stability of the object's composition
If transgenic mice overexpressing A30P alpha-S are used, then aggregation delay is observed, but neurodegeneration does not occur
Solution Approach 1:
The patent changes the critical parameter of amino acid substitution from A30P to A56P. This parameter change fundamentally alters the protein's behavior, producing a mutant that both delays aggregation kinetics and generates neurotoxicity, thereby resolving the contradiction observed with A30P mutants.
Data Source
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AI summary
The present invention relates to a mutant human alpha-synuclein with increased toxicity compared to wild-type alpha-synuclein, or a homologue thereof, wherein the mutant alpha-synuclein or homologue thereof comprises at least one amino acid substitution selected from the group consisting of a substitution at the alanine at position 56 (A56), at the alanine at position 76 (A76), at the methionine at position 127 (M 127) and/or at the valine at position 118 (V118), as defined in the claims. Further, the invention relates to a polynucleotide encoding the mutant alpha-synuclein or homologue thereof, or an expression vector comprising said polynucleotide, a cell comprising the polynucleotide or expression vector, as defined in the claims. Also, a non-human animal comprising the cell of the invention is provided, as defined in the claims. Finally, the invention provides methods for identifying a substance that prevents or reduces toxicity of alpha-synuclein, as defined in the claims.