Refining α-synuclein seed amplification assays to distinguish Parkinson’s disease from multiple system atrophy DOI Creative Commons
James A. Wiseman, Clinton Turner, Richard L. M. Faull

et al.

Translational Neurodegeneration, Journal Year: 2025, Volume and Issue: 14(1)

Published: Feb. 7, 2025

Abstract Background Parkinson’s disease (PD) and multiple system atrophy (MSA) are two distinct α-synucleinopathies traditionally differentiated through clinical symptoms. Early diagnosis of MSA is problematic, seed amplification assays (SAAs), such as real-time quaking-induced conversion (RT-QuIC), offer the potential to distinguish these diseases their underlying α-synuclein (α-Syn) pathology proteoforms. Currently, SAAs provide a binary result, signifying either presence or absence α-Syn seeds. To enhance diagnostic biological relevance assays, there pressing need incorporate quantification stratification proteoform-specific aggregation kinetics into current SAA pipelines. Methods Optimal RT-QuIC assay conditions for seeds extracted from PD patient brains were determined, assessed different pathologically relevant brain regions (medulla, substantia nigra, hippocampus, middle temporal gyrus, cerebellum). The conformational profiles disease- region-specific proteoforms determined by subjecting amplified reaction products concentration-dependent proteolytic digestion with proteinase K. Results Using our protocol, could be accurately delineated using kinetics, including rate, maximum relative fluorescence, gradient amplification, core protofilament size. cases yielded significantly higher values than across all four kinetic parameters in tissues, MSA-cerebellar phenotype having fluorescence MSA-Parkinsonian phenotype. Statistical significance was maintained when data analysed regionally grouped. Conclusions Our protocol analysis pipeline can between MSA, phenotypes. induce faster propagation exhibit α-Syn, mirroring differences observed tissue. With further validation quantitative parameters, we propose that advance yes/no theranostic biomarker utilised developing therapeutics.

Language: Английский

Refining α-synuclein seed amplification assays to distinguish Parkinson’s disease from multiple system atrophy DOI Creative Commons
James A. Wiseman, Clinton Turner, Richard L. M. Faull

et al.

Translational Neurodegeneration, Journal Year: 2025, Volume and Issue: 14(1)

Published: Feb. 7, 2025

Abstract Background Parkinson’s disease (PD) and multiple system atrophy (MSA) are two distinct α-synucleinopathies traditionally differentiated through clinical symptoms. Early diagnosis of MSA is problematic, seed amplification assays (SAAs), such as real-time quaking-induced conversion (RT-QuIC), offer the potential to distinguish these diseases their underlying α-synuclein (α-Syn) pathology proteoforms. Currently, SAAs provide a binary result, signifying either presence or absence α-Syn seeds. To enhance diagnostic biological relevance assays, there pressing need incorporate quantification stratification proteoform-specific aggregation kinetics into current SAA pipelines. Methods Optimal RT-QuIC assay conditions for seeds extracted from PD patient brains were determined, assessed different pathologically relevant brain regions (medulla, substantia nigra, hippocampus, middle temporal gyrus, cerebellum). The conformational profiles disease- region-specific proteoforms determined by subjecting amplified reaction products concentration-dependent proteolytic digestion with proteinase K. Results Using our protocol, could be accurately delineated using kinetics, including rate, maximum relative fluorescence, gradient amplification, core protofilament size. cases yielded significantly higher values than across all four kinetic parameters in tissues, MSA-cerebellar phenotype having fluorescence MSA-Parkinsonian phenotype. Statistical significance was maintained when data analysed regionally grouped. Conclusions Our protocol analysis pipeline can between MSA, phenotypes. induce faster propagation exhibit α-Syn, mirroring differences observed tissue. With further validation quantitative parameters, we propose that advance yes/no theranostic biomarker utilised developing therapeutics.

Language: Английский

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