Ultrasensitive Detection of Aggregated α-Synuclein in Glial Cells, Human Cerebrospinal Fluid, and Brain Tissue Using the RT-QuIC Assay: New High-Throughput Neuroimmune Biomarker Assay for Parkinsonian Disorders
| العنوان: | Ultrasensitive Detection of Aggregated α-Synuclein in Glial Cells, Human Cerebrospinal Fluid, and Brain Tissue Using the RT-QuIC Assay: New High-Throughput Neuroimmune Biomarker Assay for Parkinsonian Disorders |
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| المؤلفون: | Huajun Jin, Anumantha G. Kanthasamy, Monica Hepker, Mechelle M. Lewis, Vellareddy Anantharam, Naveen Kondru, Xuemei Huang, Arthi Kanthasamy, Sireesha Manne |
| المصدر: | J Neuroimmune Pharmacol |
| بيانات النشر: | Springer Science and Business Media LLC, 2019. |
| سنة النشر: | 2019 |
| مصطلحات موضوعية: | 0301 basic medicine, Synucleinopathies, Protein aggregation, law.invention, Mice, 0302 clinical medicine, Cerebrospinal fluid, law, Immunology and Allergy, Fluorometry, Single-Blind Method, Aged, 80 and over, Microglia, Molecular pathology, Chemistry, Age Factors, Middle Aged, Recombinant Proteins, medicine.anatomical_structure, alpha-Synuclein, Recombinant DNA, Biomarker (medicine), Neuroglia, Lewy Body Disease, Immunology, Neuroscience (miscellaneous), Sensitivity and Specificity, Article, Progressive supranuclear palsy, Protein Aggregates, 03 medical and health sciences, Parkinsonian Disorders, Alzheimer Disease, Computer Systems, medicine, Animals, Humans, Benzothiazoles, Aged, Fluorescent Dyes, Brain Chemistry, Pharmacology, Dementia with Lewy bodies, Reproducibility of Results, medicine.disease, Molecular biology, High-Throughput Screening Assays, 030104 developmental biology, Case-Control Studies, Biomarkers, 030217 neurology & neurosurgery |
| الوصف: | Adult-onset neurodegenerative disorders, like Parkinson’s disease (PD) and dementia with Lewy bodies (DLB), that share the accumulation of aggregated α-synuclein (αSynagg) as their hallmark molecular pathology are collectively known as α-synucleinopathies. Diagnosing α-synucleinopathies requires the post-mortem detection of αSynagg in various brain regions. Recent efforts to measure αSynagg in living patients include quantifying αSynagg 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 αSynagg. We first optimized RT-QuIC for sensitivity, specificity, and reproducibility by using monomeric recombinant human wild-type αSyn as a substrate and αSynagg as the seed. Next, we exposed mouse microglia to αSyn pre-formed fibrils (αSynPFF) for 24 h. RT-QuIC assay revealed that the αSynPFF is taken up rapidly by mouse microglia, within 30 min, and cleared within 24 h. We then evaluated the αSyn RT-QuIC assay for detecting αSynagg 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 αSynagg. The PD and DLB BH samples not only showed significantly higher αSynagg 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 αSynagg PAR compared to age-matched healthy controls, with 100% sensitivity and specificity. Overall, the RT-QuIC assay accurately detects αSynagg seeding activity, offering a potential tool for antemortem diagnosis of α-synucleinopathies and other protein-misfolding disorders. |
| تدمد: | 1557-1904 1557-1890 |
| الوصول الحر: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::fe52c0c96979fe3a195e6430a048e8f8Test https://doi.org/10.1007/s11481-019-09835-4Test |
| حقوق: | OPEN |
| رقم الانضمام: | edsair.doi.dedup.....fe52c0c96979fe3a195e6430a048e8f8 |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 15571904 15571890 |
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