Journal
JOURNAL OF NEUROIMMUNE PHARMACOLOGY
Volume 14, Issue 3, Pages 423-435Publisher
SPRINGER
DOI: 10.1007/s11481-019-09835-4
Keywords
Biomarker; alpha-synuclein; alpha-synucleinopathy; RT-QuIC assay; Neuroimmune; Diagnosis; Protein aggregation
Categories
Funding
- NIH/NIA [P30 AG10129]
- National Institutes of Health [ES026892, NS100090, NS088206]
- Presidential Interdisciplinary Research Initiative for the Big Data Brain Research from Iowa State University
- National Center for Advancing Translational Sciences [UL1 TR000127, UL1 RR033184, NS060722, NS082151, TL1 TR002016]
- PA Department of Health Tobacco CURE Funds
- Translational Brain Research Center
- Michael J. Fox Foundation for Parkinson's Research
- Alzheimer's Association
- Alzheimer's Research UK
- Weston Brain Institute
Ask authors/readers for more resources
Adult-onset neurodegenerative disorders, like Parkinson's disease (PD) and dementia with Lewy bodies (DLB), that share the accumulation of aggregated alpha-synuclein (alpha Syn(agg)) as their hallmark molecular pathology are collectively known as alpha-synucleinopathies. Diagnosing alpha-synucleinopathies requires the post-mortem detection of alpha Syn(agg) in various brain regions. Recent efforts to measure alpha Syn(agg) in living patients include quantifying alpha Syn(agg) in different biofluids as a biomarker for PD. We adopted the real-time quaking-induced conversion (RT-QuIC) assay to detect very low levels of alpha Syn(agg). We first optimized RT-QuIC for sensitivity, specificity, and reproducibility by using monomeric recombinant human wild-type alpha Syn as a substrate and alpha Syn(agg) as the seed. Next, we exposed mouse microglia to alpha Syn pre-formed fibrils (alpha Syn(PFF)) for 24 h. RT-QuIC assay revealed that the alpha Syn(PFF) is taken up rapidly by mouse microglia, within 30 min, and cleared within 24 h. We then evaluated the alpha Syn RT-QuIC assay for detecting alpha Syn(agg) in human PD, DLB, and Alzheimer's disease (AD) post-mortem brain homogenates (BH) along with PD and progressive supranuclear palsy (PSP) cerebrospinal fluid (CSF) samples and then determined protein aggregation rate (PAR) for alpha Syn(agg). The PD and DLB BH samples not only showed significantly higher alpha Syn(agg) PAR compared to age-matched healthy controls and AD, but RT-QuIC was also highly reproducible with 94% sensitivity and 100% specificity. Similarly, PD CSF samples demonstrated significantly higher alpha Syn(agg) PAR compared to age-matched healthy controls, with 100% sensitivity and specificity. Overall, the RT-QuIC assay accurately detects alpha Syn(agg) seeding activity, offering a potential tool for antemortem diagnosis of alpha-synucleinopathies and other protein-misfolding disorders.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available