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Antibody-free measurement of cerebrospinal fluid tau phosphorylation across the Alzheimer's disease continuum

Gobom, Johan ; Benedet, Andréa L ; Mattsson-Carlgren, Niklas LU orcid ; Montoliu-Gaya, Laia ; Schultz, Nina LU ; Ashton, Nicholas J ; Janelidze, Shorena LU ; Servaes, Stijn ; Sauer, Mathias and Pascoal, Tharick A , et al. (2022) In Molecular Neurodegeneration 17(1).
Abstract

BACKGROUND: Alzheimer's disease is characterized by an abnormal increase of phosphorylated tau (pTau) species in the CSF. It has been suggested that emergence of different pTau forms may parallel disease progression. Therefore, targeting multiple specific pTau forms may allow for a deeper understanding of disease evolution and underlying pathophysiology. Current immunoassays measure pTau epitopes separately and may capture phosphorylated tau fragments of different length depending on the non-pTau antibody used in the assay sandwich pair, which bias the measurement.

METHODS: We developed the first antibody-free mass spectrometric method to simultaneously measure multiple phosphorylated epitopes in CSF tau: pT181, pS199, pS202,... (More)

BACKGROUND: Alzheimer's disease is characterized by an abnormal increase of phosphorylated tau (pTau) species in the CSF. It has been suggested that emergence of different pTau forms may parallel disease progression. Therefore, targeting multiple specific pTau forms may allow for a deeper understanding of disease evolution and underlying pathophysiology. Current immunoassays measure pTau epitopes separately and may capture phosphorylated tau fragments of different length depending on the non-pTau antibody used in the assay sandwich pair, which bias the measurement.

METHODS: We developed the first antibody-free mass spectrometric method to simultaneously measure multiple phosphorylated epitopes in CSF tau: pT181, pS199, pS202, pT205, pT217, pT231, and pS396. The method was first evaluated in biochemically defined Alzheimer's disease and control CSF samples (n = 38). All seven pTau epitopes clearly separated Alzheimer's disease from non-AD (p < 0.001, AUC = 0.84-0.98). We proceeded with clinical validation of the method in the TRIAD (n = 165) and BioFINDER-2 cohorts (n = 563), consisting of patients across the full Alzheimer's disease continuum, including also young controls (< 40 years), as well as patients with frontotemporal dementia and other neurodegenerative disorders.

RESULTS: Increased levels of all phosphorylated epitopes were found in Alzheimer's disease dementia and Aβ positron emission tomography-positive patients with mild cognitive impairment compared with Aβ-negative controls. For Alzheimer's disease dementia compared with Aβ-negative controls, the best biomarker performance was observed for pT231 (TRIAD: AUC = 98.73%, fold change = 7.64; BioFINDER-2: AUC = 91.89%, fold change = 10.65), pT217 (TRIAD: AUC = 99.71%, fold change = 6.33; BioFINDER-2: AUC = 98.12%, fold change = 8.83) and pT205 (TRIAD: AUC = 99.07%, fold change = 5.34; BioFINDER-2: AUC = 93.51%, fold change = 3.92). These phospho-epitopes also discriminated between Aβ-positive and Aβ-negative cognitively unimpaired individuals: pT217 (TRIAD: AUC = 83.26, fold change = 2.39; BioFINDER-2: AUC = 91.05%, fold change = 3.29), pT231 (TRIAD: AUC = 86.25, fold change = 3.80; BioFINDER-2: AUC = 78.69%, fold change = 3.65) and pT205 (TRIAD: AUC = 71.58, fold change = 1.51; BioFINDER-2: AUC = 71.11%, fold change = 1.70).

CONCLUSIONS: While an increase was found for all pTau species examined, the highest fold change in Alzheimer's disease was found for pT231, pT217 and pT205. Simultaneous antibody-free measurement of pTau epitopes by mass spectrometry avoids possible bias caused by differences in antibody affinity for modified or processed forms of tau, provides insights into tau pathophysiology and may facilitate clinical trials on tau-based drug candidates.

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type
Contribution to journal
publication status
published
subject
keywords
Humans, Alzheimer Disease/cerebrospinal fluid, Amyloid beta-Peptides/cerebrospinal fluid, tau Proteins/metabolism, Phosphorylation, Cognitive Dysfunction, Biomarkers/cerebrospinal fluid, Peptide Fragments/cerebrospinal fluid
in
Molecular Neurodegeneration
volume
17
issue
1
article number
81
publisher
BioMed Central (BMC)
external identifiers
  • pmid:36510321
  • scopus:85144340704
ISSN
1750-1326
DOI
10.1186/s13024-022-00586-0
language
English
LU publication?
yes
id
1be1c184-44e2-46bb-867b-bd2f7b007076
date added to LUP
2022-12-21 14:59:28
date last changed
2024-06-13 15:03:53
@article{1be1c184-44e2-46bb-867b-bd2f7b007076,
  abstract     = {{<p>BACKGROUND: Alzheimer's disease is characterized by an abnormal increase of phosphorylated tau (pTau) species in the CSF. It has been suggested that emergence of different pTau forms may parallel disease progression. Therefore, targeting multiple specific pTau forms may allow for a deeper understanding of disease evolution and underlying pathophysiology. Current immunoassays measure pTau epitopes separately and may capture phosphorylated tau fragments of different length depending on the non-pTau antibody used in the assay sandwich pair, which bias the measurement.</p><p>METHODS: We developed the first antibody-free mass spectrometric method to simultaneously measure multiple phosphorylated epitopes in CSF tau: pT181, pS199, pS202, pT205, pT217, pT231, and pS396. The method was first evaluated in biochemically defined Alzheimer's disease and control CSF samples (n = 38). All seven pTau epitopes clearly separated Alzheimer's disease from non-AD (p &lt; 0.001, AUC = 0.84-0.98). We proceeded with clinical validation of the method in the TRIAD (n = 165) and BioFINDER-2 cohorts (n = 563), consisting of patients across the full Alzheimer's disease continuum, including also young controls (&lt; 40 years), as well as patients with frontotemporal dementia and other neurodegenerative disorders.</p><p>RESULTS: Increased levels of all phosphorylated epitopes were found in Alzheimer's disease dementia and Aβ positron emission tomography-positive patients with mild cognitive impairment compared with Aβ-negative controls. For Alzheimer's disease dementia compared with Aβ-negative controls, the best biomarker performance was observed for pT231 (TRIAD: AUC = 98.73%, fold change = 7.64; BioFINDER-2: AUC = 91.89%, fold change = 10.65), pT217 (TRIAD: AUC = 99.71%, fold change = 6.33; BioFINDER-2: AUC = 98.12%, fold change = 8.83) and pT205 (TRIAD: AUC = 99.07%, fold change = 5.34; BioFINDER-2: AUC = 93.51%, fold change = 3.92). These phospho-epitopes also discriminated between Aβ-positive and Aβ-negative cognitively unimpaired individuals: pT217 (TRIAD: AUC = 83.26, fold change = 2.39; BioFINDER-2: AUC = 91.05%, fold change = 3.29), pT231 (TRIAD: AUC = 86.25, fold change = 3.80; BioFINDER-2: AUC = 78.69%, fold change = 3.65) and pT205 (TRIAD: AUC = 71.58, fold change = 1.51; BioFINDER-2: AUC = 71.11%, fold change = 1.70).</p><p>CONCLUSIONS: While an increase was found for all pTau species examined, the highest fold change in Alzheimer's disease was found for pT231, pT217 and pT205. Simultaneous antibody-free measurement of pTau epitopes by mass spectrometry avoids possible bias caused by differences in antibody affinity for modified or processed forms of tau, provides insights into tau pathophysiology and may facilitate clinical trials on tau-based drug candidates.</p>}},
  author       = {{Gobom, Johan and Benedet, Andréa L and Mattsson-Carlgren, Niklas and Montoliu-Gaya, Laia and Schultz, Nina and Ashton, Nicholas J and Janelidze, Shorena and Servaes, Stijn and Sauer, Mathias and Pascoal, Tharick A and Karikari, Thomas K and Lantero-Rodriguez, Juan and Brinkmalm, Gunnar and Zetterberg, Henrik and Hansson, Oskar and Rosa-Neto, Pedro and Blennow, Kaj}},
  issn         = {{1750-1326}},
  keywords     = {{Humans; Alzheimer Disease/cerebrospinal fluid; Amyloid beta-Peptides/cerebrospinal fluid; tau Proteins/metabolism; Phosphorylation; Cognitive Dysfunction; Biomarkers/cerebrospinal fluid; Peptide Fragments/cerebrospinal fluid}},
  language     = {{eng}},
  month        = {{12}},
  number       = {{1}},
  publisher    = {{BioMed Central (BMC)}},
  series       = {{Molecular Neurodegeneration}},
  title        = {{Antibody-free measurement of cerebrospinal fluid tau phosphorylation across the Alzheimer's disease continuum}},
  url          = {{http://dx.doi.org/10.1186/s13024-022-00586-0}},
  doi          = {{10.1186/s13024-022-00586-0}},
  volume       = {{17}},
  year         = {{2022}},
}