Skip to main content
Key Specialties
Cardiology Diabetology Endocrinology Gastroenterology Geriatrics and Gerontology Gynecology and Obstetrics Hematology Infectious Disease Internal Medicine Nephrology Neurology Oncology Pediatrics Respiratory Medicine Rheumatology Surgery
Congress Coverage
2024 ACC Congress 2023 ESMO Congress 2023 EASD Congress 2023 ERS Congress 2023 ESC Congress
Clinical Collections
Advances in Alzheimer’s

Keynote webinar | Spotlight on medication adherence

LIVE: Thursday 27th June 2024, 18:00-19:30 (CEST)
Join our expert panel to discover why you need to understand the drivers of non-adherence in your patients, and how you can optimize medication adherence in your clinics to drastically improve patient outcomes.
ADVANCED SEARCH
Log in
Top
Acta Neuropathologica
Published in: Acta Neuropathologica 1/2020

Open Access 01-07-2020 | Alzheimer's Disease | Original Paper

Ultrasensitive RT-QuIC assay with high sensitivity and specificity for Lewy body-associated synucleinopathies

Authors: Marcello Rossi, Niccolò Candelise, Simone Baiardi, Sabina Capellari, Giulia Giannini, Christina D. Orrù, Elena Antelmi, Angela Mammana, Andrew G. Hughson, Giovanna Calandra-Buonaura, Anna Ladogana, Giuseppe Plazzi, Pietro Cortelli, Byron Caughey, Piero Parchi

Published in: Acta Neuropathologica | Issue 1/2020

Login to get access

Abstract

The clinical diagnosis of synucleinopathies, including Parkinson’s disease (PD), dementia with Lewy bodies (DLB), and multiple system atrophy (MSA), is challenging, especially at an early disease stage, due to the heterogeneous and often non-specific clinical manifestations. The discovery of reliable specific markers for synucleinopathies would consequently be of great aid to the diagnosis and management of these disorders. Real-Time Quaking-Induced Conversion (RT-QuIC) is an ultrasensitive technique that has been previously used to detect self-templating amyloidogenic proteins in the cerebrospinal fluid (CSF) and other biospecimens in prion disease and synucleinopathies. Using a wild-type recombinant α-synuclein as a substrate, we applied RT-QuIC to a large cohort of 439 CSF samples from clinically well-characterized, or post-mortem verified patients with parkinsonism or dementia. Of significance, we also studied patients with isolated REM sleep behavior disorder (iRBD) (n = 18) and pure autonomic failure (PAF) (n = 28), representing clinical syndromes that are often caused by a synucleinopathy, and may precede the appearance of parkinsonism or cognitive decline. The results show that our RT-QuIC assay can accurately detect α-synuclein seeding activity across the spectrum of Lewy Body (LB)-related disorders (LBD), including DLB, PD, iRBD, and PAF, with an overall sensitivity of 95.3%. In contrast, all but two patients with MSA showed no α-synuclein seeding activity in the applied experimental setting. The analysis of the fluorescence response reflecting the amount of α-synuclein seeds revealed no significant differences between the clinical syndromes associated with LB pathology. Finally, the assay demonstrated 98% specificity in a neuropathological cohort of 101 cases lacking LB pathology. In conclusion, α-synuclein RT-QuIC provides an accurate marker of synucleinopathies linked to LB pathology and may have a pivotal role in the early discrimination and management of affected patients. The finding of no α-synuclein seeding activity in MSA seems to support the current view that MSA and LBD are associated with different conformational strains of α-synuclein.
Appendix
Available only for authorised users
Literature
1.
Alafuzoff I, Arzberger T, Al-Sarraj S, Bodi I, Bogdanovic N, Braak H et al (2008) Staging of neurofibrillary pathology in Alzheimer's disease: a study of the BrainNet Europe Consortium. Brain Pathol 18(4):484–496PubMedPubMedCentral
2.
Alafuzoff I, Ince PG, Arzberger T, Al-Sarraj S, Bell J, Bodi I et al (2009) Staging/typing of Lewy body related alpha-synuclein pathology: a study of the BrainNet Europe Consortium. Acta Neuropathol 117(6):635–652CrossRefPubMed
3.
Armstrong MJ, Litvan I, Lang AE, Bak TH, Bhatia KP, Borroni B et al (2013) Criteria for the diagnosis of corticobasal degeneration. Neurology 80(5):496–503PubMedPubMedCentralCrossRef
4.
Atarashi R, Satoh K, Sano K, Fuse T, Yamaguchi N, Ishibashi D et al (2011) Ultrasensitive human prion detection in cerebrospinal fluid by real-time quaking-induced conversion. Nat Med 17(2):175–178CrossRefPubMed
5.
Baiardi S, Abu-Rumeileh S, Rossi M, Zenesini C, Bartoletti-Stella A, Polischi B et al (2018) Antemortem CSF Aβ42/Aβ40 ratio predicts Alzheimer's disease pathology better than Aβ42 in rapidly progressive dementias. Ann Clin Transl Neurol 6(2):263–273PubMedPubMedCentral
6.
Baiardi S, Redaelli V, Ripellino P, Rossi M, Franceschini A, Moggio M et al (2019) Prion-related peripheral neuropathy in sporadic Creutzfeldt-Jakob disease. J Neurol Neurosurg Psychiatry 90(4):424–427CrossRefPubMed
7.
Barker RA, Williams-Gray CH (2016) Review: The spectrum of clinical features seen with alpha synuclein pathology. Neuropathol Appl Neurobiol 42(1):6–19CrossRefPubMed
8.
Bloch A, Probst A, Bissig H, Adams H, Tolnay M (2006) Alpha-synuclein pathology of the spinal and peripheral autonomic nervous system in neurologically unimpaired elderly subjects. Neuropathol Appl Neurobiol 32(3):284–295CrossRefPubMed
9.
Bongianni M, Ladogana A, Capaldi S, Klotz S, Baiardi S, Cagnin A et al (2019) α-Synuclein RT-QuIC assay in cerebrospinal fluid of patients with dementia with Lewy bodies. Ann Clin Transl Neurol 6(10):2120–2126PubMedPubMedCentralCrossRef
10.
Bousset L, Pieri L, Ruiz-Arlandis G, Gath J, Jensen PH, Habenstein B et al (2013) Structural and functional characterization of two alpha-synuclein strains. Nat Commun 4:2575PubMedCrossRef
11.
Braune S, Reinhardt M, Schnitzer R, Riedel A, Lücking CH (1999) Cardiac uptake of [123I]MIBG separates Parkinson's disease from multiple system atrophy. Neurology 53(5):1020–1025CrossRefPubMed
12.
Candelise N, Schmitz M, Llorens F, Villar-Piqué A, Cramm M, Thom T et al (2019) Seeding variability of different alpha synuclein strains in synucleinopathies. Ann Neurol 85(5):691–703CrossRefPubMed
13.
Chandler MP, Mathias CJ (2002) Haemodynamic responses during head-up tilt and tilt reversal in two groups with chronic autonomic failure: pure autonomic failure and multiple system atrophy. J Neurol 249(5):542–548CrossRefPubMed
14.
Colby DW, Zhang Q, Wang S, Groth D, Legname G, Riesner D et al (2007) Prion detection by an amyloid seeding assay. Proc Natl Acad Sci USA 104(52):20914–20919CrossRefPubMedPubMedCentral
15.
Concha-Marambio L, Shahnawaz M, Soto C (2019) Detection of misfolded α-synuclein aggregates in cerebrospinal fluid by the protein misfolding cyclic amplification platform. Methods Mol Biol 1948:35–44CrossRefPubMed
16.
17.
Cramm M, Schmitz M, Karch A, Mitrova E, Kuhn F, Schroeder B et al (2016) Stability and reproducibility underscore utility of RT-QuIC for diagnosis of Creutzfeldt–Jakob disease. Mol Neurobiol 53(3):1896–1904CrossRefPubMed
18.
Crary JF, Trojanowski JQ, Schneider JA, Abisambra JF, Abner EL, Alafuzoff I et al (2014) Primary age-related tauopathy (PART): a common pathology associated with human aging. Acta Neuropathol 128(6):755–766PubMedPubMedCentralCrossRef
19.
Dubois B, Feldman HH, Jacova C, Hampel H, Molinuevo JL, Blennow K et al (2014) Advancing research diagnostic criteria for Alzheimer's disease: the IWG-2 criteria. Lancet Neurol 13(6):614–629CrossRefPubMed
20.
Fairfoul G, McGuire LI, Pal S, Ironside JW, Neumann J, Christie S et al (2016) Alpha-synuclein RT-QuIC in the CSF of patients with alpha-synucleinopathies. Ann Clin Transl Neurol 3(10):812–818PubMedPubMedCentralCrossRef
21.
Foutz A, Appleby BS, Hamlin C, Liu X, Yang S, Cohen Y et al (2017) Diagnostic and prognostic value of human prion detection in cerebrospinal fluid. Ann Neurol 81(1):79–92PubMedPubMedCentralCrossRef
22.
Franceschini A, Baiardi S, Hughson AG, McKenzie N, Moda F, Rossi M et al (2017) High diagnostic value of second generation CSF RT-QuIC across the wide spectrum of CJD prions. Sci Rep 7(1):10655PubMedPubMedCentralCrossRef
23.
24.
Garrido A, Fairfoul G, Tolosa ES, Martí MJ, Green A, Barcelona LRRK2 Study Group (2019) α-synuclein RT-QuIC in cerebrospinal fluid of LRRK2-linked Parkinson's disease. Ann Clin Transl Neurol 6(6):1024–1032PubMedPubMedCentral
25.
Giannini G, Calandra-Buonaura G, Asioli GM, Cecere A, Barletta G, Mignani F et al (2018) The natural history of idiopathic autonomic failure: the IAF-BO cohort study. Neurology 91(13):e1245–1254CrossRefPubMed
26.
Gilman S, Wenning GK, Low PA, Brooks DJ, Mathias CJ, Trojanowski JQ et al (2008) Second consensus statement on the diagnosis of multiple system atrophy. Neurology 71(9):670–676PubMedPubMedCentralCrossRef
27.
Goedert M, Masuda-Suzukake M, Falcon B (2017) Like prions: the propagation of aggregated tau and α-synuclein in neurodegeneration. Brain 140(2):266–278CrossRefPubMed
28.
Groveman BR, Orrù CD, Hughson AG, Raymond LD, Zanusso G, Ghetti B et al (2018) Rapid and ultra-sensitive quantitation of disease-associated α-synuclein seeds in brain and cerebrospinal fluid by αSyn RT-QuIC. Acta Neuropathol Commun 6(1):7PubMedPubMedCentralCrossRef
29.
Halliday GM, Holton JL, Revesz T, Dickson DW (2011) Neuropathology underlying clinical variability in patients with synucleinopathies. Acta Neuropathol 122(2):187–204CrossRefPubMed
30.
Hogl B, Stefani A, Videnovic A (2018) Idiopathic REM sleep behaviour disorder and neurodegeneration—an update. Nat Rev Neurol 14(1):40–55CrossRefPubMed
31.
Höglinger GU, Respondek G, Stamelou M, Kurz C, Josephs KA, Lang AE et al (2017) Clinical diagnosis of progressive supranuclear palsy: the movement disorder society criteria. Mov Disord 32(6):853–864PubMedPubMedCentralCrossRef
32.
Iranzo A, Fernández-Arcos A, Tolosa E, Serradell M, Molinuevo JL, Valldeoriola F et al (2014) Neurodegenerative disorder risk in idiopathic REM sleep behavior disorder: study in 174 patients. PLoS ONE 9(2):e89741PubMedPubMedCentralCrossRef
33.
Iranzo A, Santamaria J, Tolosa E (2016) Idiopathic rapid eye movement sleep behaviour disorder: diagnosis, management, and the need for neuroprotective interventions. Lancet Neurol 15(4):405–419CrossRefPubMed
34.
35.
Kaufmann H, Norcliffe-Kaufmann L, Palma JA, Biaggioni I, Low PA, Singer W et al (2017) Natural history of pure autonomic failure: a United States prospective cohort. Ann Neurol 81(2):287–297PubMedPubMedCentralCrossRef
36.
Klos KJ, Ahlskog JE, Josephs KA et al (2006) Alpha-synuclein pathology in the spinal cords of neurologically asymptomatic aged individuals. Neurology 66(7):1100–1102CrossRefPubMed
37.
Lattanzio F, Abu-Rumeileh S, Franceschini A, Kai H, Amore G, Poggiolini I et al (2017) Prion-specific and surrogate CSF biomarkers in Creutzfeldt-Jakob disease: diagnostic accuracy in relation to molecular subtypes and analysis of neuropathological correlates of p-tau and Aβ42 levels. Acta Neuropathol 133(4):559–578PubMedPubMedCentralCrossRef
38.
Lau A, So RWL, Lau HHC, Sang JC, Ruiz-Riquelme A, Fleck SC et al (2020) α-Synuclein strains target distinct brain regions and cell types. Nat Neurosci 23(1):21–31PubMedCrossRef
39.
40.
McGuire LI, Peden AH, Orrú CD, Wilham JM, Appleford NE, Mallinson G et al (2012) Real time quaking-induced conversion analysis of cerebrospinal fluid in sporadic Creutzfeldt–Jakob disease. Ann Neurol 72(2):278–285PubMedPubMedCentralCrossRef
41.
McKeith IG, Boeve BF, Dickson DW, Halliday G, Taylor JP, Weintraub D et al (2017) Diagnosis and management of dementia with Lewy bodies: fourth consensus report of the DLB Consortium. Neurology 89(1):88–100PubMedPubMedCentralCrossRef
42.
Meissner WG, Fernagut PO, Dehay B, Péran P, Traon AP, Foubert-Samier A et al (2019) Multiple system atrophy: recent developments and future perspectives. Mov Disord 34(11):1629–1642CrossRefPubMed
43.
Montine TJ, Phelps CH, Beach TG, Bigio EH, Cairns NJ, Dickson DW et al (2012) National Institute on Aging-Alzheimer's Association guidelines for the neuropathologic assessment of Alzheimer's disease: a practical approach. Acta Neuropathol 123(1):1–11PubMedCrossRef
44.
Orrú CD, Groveman BR, Raymond LD, Hughson AG, Nonno R, Zou W et al (2015) Bank vole prion protein as an apparently universal substrate for RT-QuIC-based detection and discrimination of prion strains. PLoS Pathog 11(86):e1004983PubMedPubMedCentralCrossRef
45.
Orrú CD, Groveman BR, Hughson AG, Zanusso G, Coulthart MB, Caughey B (2015) Rapid and sensitive RT-QuIC detection of human Creutzfeldt–Jakob disease using cerebrospinal fluid. mBio 6(1):e02451–e2514PubMedPubMedCentralCrossRef
46.
Parchi P, de Boni L, Saverioni D, Cohen ML, Ferrer I, Gambetti P et al (2012) Consensus classification of human prion disease histotypes allows reliable identification of molecular subtypes: an inter-rater study among surveillance centres in Europe and USA. Acta Neuropathol 124(4):517–529PubMedPubMedCentralCrossRef
47.
Parchi P, Strammiello R, Notari S, Giese A, Langeveld JP, Ladogana A et al (2009) Incidence and spectrum of sporadic Creutzfeldt–Jakob disease variants with mixed phenotype and co-occurrence of PrPSc types: an updated classification. Acta Neuropathol 118(5):659–671PubMedPubMedCentralCrossRef
48.
Parnetti L, Gaetani L, Eusebi P, Paciotti S, Hansson O, El-Agnaf O et al (2019) CSF and blood biomarkers for Parkinson's disease. Lancet Neurol 18(6):573–586CrossRefPubMed
49.
Peelaerts W, Bousset L, Van der Perren A, Moskalyuk A, Pulizzi R, Giugliano M et al (2015) α-Synuclein strains cause distinct synucleinopathies after local and systemic administration. Nature 522(7556):340–344CrossRefPubMed
50.
Postuma RB, Berg D, Stern M, Poewe W, Olanow CW, Oertel W et al (2015) MDS clinical diagnostic criteria for Parkinson's disease. Mov Disord 30(12):1591–1601CrossRefPubMed
51.
Postuma RB, Iranzo A, Hu M, Högl B, Boeve BF, Manni R et al (2019) Risk and predictors of dementia and parkinsonism in idiopathic REM sleep behaviour disorder: a multicentre study. Brain 142(3):744–759PubMedPubMedCentralCrossRef
52.
Prusiner SB, Woerman AL, Mordes DA, Watts JC, Rampersaud R, Berry DB et al (2015) Evidence for α-synuclein prions causing multiple system atrophy in humans with parkinsonism. Proc Natl Acad Sci USA 112(38):e5308–5317CrossRefPubMedPubMedCentral
53.
54.
Saborio GP, Permanne B, Soto C (2001) Sensitive detection of pathological prion protein by cyclic amplification of protein misfolding. Nature 411(6839):810–813CrossRefPubMed
55.
Saijo E, Groveman BR, Kraus A, Metrick M, Orrù CD, Hughson AG et al (2019) Ultrasensitive RT-QuIC seed amplification assays for disease-associated tau, α-synuclein, and prion aggregates. Methods Mol Biol 1873:19–37CrossRefPubMed
56.
Saijo E, Metrick MA 2nd, Koga S, Parchi P, Litvan I, Spina S et al (2020) 4-Repeat tau seeds and templating subtypes as brain and CSF biomarkers of frontotemporal lobar degeneration. Acta Neuropathol 139(1):63–77CrossRefPubMed
57.
Sateia MJ (2014) International classification of sleep disorders-third edition: highlights and modifications. Chest 146(5):1387–1394CrossRefPubMed
58.
Schapira AHV, Chaudhuri KR, Jenner P (2017) Non-motor features in Parkinson’s disease. Nat Rev Neurosci 18(8):509CrossRefPubMed
59.
60.
Shahnawaz M, Tokuda T, Waragai M, Mendez N, Ishii R, Trenkwalder C et al (2017) Development of a biochemical diagnosis of Parkinson disease by detection of α-synuclein misfolded aggregates in cerebrospinal fluid. JAMA Neurol 74(2):163–172CrossRefPubMed
61.
62.
Spillantini MG, Schmidt ML, Lee VM, Trojanowski JQ, Jakes R, Goedert M (1997) Alpha-synuclein in Lewy bodies. Nature 388(6645):839–840CrossRefPubMed
63.
Taki J, Yoshita M, Yamada M, Tonami N (2004) Significance of 123I-MIBG scintigraphy as a pathophysiological indicator in the assessment of Parkinson's disease and related disorders: it can be a specific marker for Lewy body disease. Ann Nucl Med 18(6):453–461CrossRefPubMed
64.
van Rumund A, Green AJE, Fairfoul G, Esselink RAJ, Bloem BR, Verbeek MM (2019) α-Synuclein real-time quaking-induced conversion in the cerebrospinal fluid of uncertain cases of parkinsonism. Ann Neurol 85(5):777–781PubMedPubMedCentralCrossRef
65.
66.
Wakisaka Y, Furuta A, Tanizaki Y, Kiyohara Y, Iida M, Iwaki T (2003) Age-associated prevalence and risk factors of Lewy body pathology in a general population: the Hisayama study. Acta Neuropathol 106(4):374–382CrossRefPubMed
67.
68.
Yamasaki TR, Holmes BB, Furman JL, Dhavale DD, Su BW, Song ES et al (2019) Parkinson's disease and multiple system atrophy have distinct α-synuclein seed characteristics. J Biol Chem 294(3):1045–1058PubMedCrossRef
69.
Zanusso G, Monaco S, Pocchiari M, Caughey B (2016) Advanced tests for early and accurate diagnosis of Creutzfeldt–Jakob disease. Nat Rev Neurol 12(6):325–333CrossRefPubMed
70.
Metadata
Title
Ultrasensitive RT-QuIC assay with high sensitivity and specificity for Lewy body-associated synucleinopathies
Authors
Marcello Rossi
Niccolò Candelise
Simone Baiardi
Sabina Capellari
Giulia Giannini
Christina D. Orrù
Elena Antelmi
Angela Mammana
Andrew G. Hughson
Giovanna Calandra-Buonaura
Anna Ladogana
Giuseppe Plazzi
Pietro Cortelli
Byron Caughey
Piero Parchi
Publication date
01-07-2020
Publisher
Springer Berlin Heidelberg
Published in
Acta Neuropathologica / Issue 1/2020
Print ISSN: 0001-6322
Electronic ISSN: 1432-0533
DOI
https://doi.org/10.1007/s00401-020-02160-8

Other articles of this Issue 1/2020

Acta Neuropathologica 1/2020 Go to the issue

Correspondence

Spatial enrichment of cellular states in glioblastoma

Correspondence

Frequent inactivating mutations of the PBAF complex gene PBRM1 in meningioma with papillary features

Original Paper

PTEN activation contributes to neuronal and synaptic engulfment by microglia in tauopathy

Correspondence

CRISPR deletion of the C9ORF72 promoter in ALS/FTD patient motor neurons abolishes production of dipeptide repeat proteins and rescues neurodegeneration

Original Paper

MEF2 impairment underlies skeletal muscle atrophy in polyglutamine disease

Original Paper

Biological sex and DNA repair deficiency drive Alzheimer’s disease via systemic metabolic remodeling and brain mitochondrial dysfunction