Phospho-S129 Alpha-Synuclein Is Present in Human Plasma but Not in Cerebrospinal Fluid as Determined by an Ultrasensitive Immunoassay

Cristina Cariulo, Paola Martufi, Margherita Verani, Lucia Azzollini, Giordana Bruni, Andreas Weiss, Sean M. Deguire, Hilal A. Lashuel, Eugenia Scaricamazza, Giulia Maria Sancesario, Tommaso Schirinzi, Nicola Biagio Mercuri, Giuseppe Sancesario, Andrea Caricasole, Lara Petricca

Research output: Contribution to journalArticle

Abstract

Accumulation and aggregation of misfolded alpha-synuclein is believed to be a cause of Parkinson’s disease (PD). Phosphorylation of alpha-synuclein at S129 is known to be associated with the pathological misfolding process, but efforts to investigate the relevance of this post-translational modification for pathology have been frustrated by difficulties in detecting and quantifying it in relevant samples. We report novel, ultrasensitive immunoassays based on single-molecule counting technology, useful for detecting alpha-synuclein and its S129 phosphorylated form in clinical samples in the low pg/ml range. Using human CSF and plasma samples, we find levels of alpha-synuclein comparable to those previously reported. However, while alpha-synuclein phosphorylated on S129 could easily be detected in human plasma, where its detection is extremely sensitive to protein phosphatases, its levels in CSF were undetectable, with a possible influence of a matrix effect. In plasma samples from a small test cohort comprising 5 PD individuals and five age-matched control individuals we find that pS129 alpha-synuclein levels are increased in PD plasma samples, in line with previous reports. We conclude that pS129 alpha-synuclein is not detectable in CSF and recommend the addition of phosphatase inhibitors to plasma samples at the time of collection. Moreover, the findings obtained on the small cohort of clinical plasma samples point to plasma pS129 alpha-synuclein levels as a candidate diagnostic biomarker in PD.

Original languageEnglish
Article number889
JournalFrontiers in Neuroscience
Volume13
DOIs
Publication statusPublished - Aug 22 2019

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alpha-Synuclein
Immunoassay
Cerebrospinal Fluid
Parkinson Disease
Phosphoprotein Phosphatases
Pathologic Processes
Post Translational Protein Processing
Phosphoric Monoester Hydrolases
Biomarkers
Phosphorylation
Pathology
Technology

Keywords

  • alpha-synuclein
  • CSF
  • human
  • immunoassay
  • neurodegeneration
  • Parkinson’s disease
  • phosphorylation
  • plasma

ASJC Scopus subject areas

  • Neuroscience(all)

Cite this

Phospho-S129 Alpha-Synuclein Is Present in Human Plasma but Not in Cerebrospinal Fluid as Determined by an Ultrasensitive Immunoassay. / Cariulo, Cristina; Martufi, Paola; Verani, Margherita; Azzollini, Lucia; Bruni, Giordana; Weiss, Andreas; Deguire, Sean M.; Lashuel, Hilal A.; Scaricamazza, Eugenia; Sancesario, Giulia Maria; Schirinzi, Tommaso; Mercuri, Nicola Biagio; Sancesario, Giuseppe; Caricasole, Andrea; Petricca, Lara.

In: Frontiers in Neuroscience, Vol. 13, 889, 22.08.2019.

Research output: Contribution to journalArticle

Cariulo, C, Martufi, P, Verani, M, Azzollini, L, Bruni, G, Weiss, A, Deguire, SM, Lashuel, HA, Scaricamazza, E, Sancesario, GM, Schirinzi, T, Mercuri, NB, Sancesario, G, Caricasole, A & Petricca, L 2019, 'Phospho-S129 Alpha-Synuclein Is Present in Human Plasma but Not in Cerebrospinal Fluid as Determined by an Ultrasensitive Immunoassay', Frontiers in Neuroscience, vol. 13, 889. https://doi.org/10.3389/fnins.2019.00889
Cariulo, Cristina ; Martufi, Paola ; Verani, Margherita ; Azzollini, Lucia ; Bruni, Giordana ; Weiss, Andreas ; Deguire, Sean M. ; Lashuel, Hilal A. ; Scaricamazza, Eugenia ; Sancesario, Giulia Maria ; Schirinzi, Tommaso ; Mercuri, Nicola Biagio ; Sancesario, Giuseppe ; Caricasole, Andrea ; Petricca, Lara. / Phospho-S129 Alpha-Synuclein Is Present in Human Plasma but Not in Cerebrospinal Fluid as Determined by an Ultrasensitive Immunoassay. In: Frontiers in Neuroscience. 2019 ; Vol. 13.
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abstract = "Accumulation and aggregation of misfolded alpha-synuclein is believed to be a cause of Parkinson’s disease (PD). Phosphorylation of alpha-synuclein at S129 is known to be associated with the pathological misfolding process, but efforts to investigate the relevance of this post-translational modification for pathology have been frustrated by difficulties in detecting and quantifying it in relevant samples. We report novel, ultrasensitive immunoassays based on single-molecule counting technology, useful for detecting alpha-synuclein and its S129 phosphorylated form in clinical samples in the low pg/ml range. Using human CSF and plasma samples, we find levels of alpha-synuclein comparable to those previously reported. However, while alpha-synuclein phosphorylated on S129 could easily be detected in human plasma, where its detection is extremely sensitive to protein phosphatases, its levels in CSF were undetectable, with a possible influence of a matrix effect. In plasma samples from a small test cohort comprising 5 PD individuals and five age-matched control individuals we find that pS129 alpha-synuclein levels are increased in PD plasma samples, in line with previous reports. We conclude that pS129 alpha-synuclein is not detectable in CSF and recommend the addition of phosphatase inhibitors to plasma samples at the time of collection. Moreover, the findings obtained on the small cohort of clinical plasma samples point to plasma pS129 alpha-synuclein levels as a candidate diagnostic biomarker in PD.",
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AU - Verani, Margherita

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AU - Bruni, Giordana

AU - Weiss, Andreas

AU - Deguire, Sean M.

AU - Lashuel, Hilal A.

AU - Scaricamazza, Eugenia

AU - Sancesario, Giulia Maria

AU - Schirinzi, Tommaso

AU - Mercuri, Nicola Biagio

AU - Sancesario, Giuseppe

AU - Caricasole, Andrea

AU - Petricca, Lara

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N2 - Accumulation and aggregation of misfolded alpha-synuclein is believed to be a cause of Parkinson’s disease (PD). Phosphorylation of alpha-synuclein at S129 is known to be associated with the pathological misfolding process, but efforts to investigate the relevance of this post-translational modification for pathology have been frustrated by difficulties in detecting and quantifying it in relevant samples. We report novel, ultrasensitive immunoassays based on single-molecule counting technology, useful for detecting alpha-synuclein and its S129 phosphorylated form in clinical samples in the low pg/ml range. Using human CSF and plasma samples, we find levels of alpha-synuclein comparable to those previously reported. However, while alpha-synuclein phosphorylated on S129 could easily be detected in human plasma, where its detection is extremely sensitive to protein phosphatases, its levels in CSF were undetectable, with a possible influence of a matrix effect. In plasma samples from a small test cohort comprising 5 PD individuals and five age-matched control individuals we find that pS129 alpha-synuclein levels are increased in PD plasma samples, in line with previous reports. We conclude that pS129 alpha-synuclein is not detectable in CSF and recommend the addition of phosphatase inhibitors to plasma samples at the time of collection. Moreover, the findings obtained on the small cohort of clinical plasma samples point to plasma pS129 alpha-synuclein levels as a candidate diagnostic biomarker in PD.

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