Development of immunoprecipitation – two-dimensional liquid chromatography – mass spectrometry methodology as biomarker read-out to quantify phosphorylated tau in cerebrospinal fluid from Alzheimer disease patients

Bijttebier, Sebastiaan; Theunis, Clara; Jahouh, Farid; Martins, Dina Rodrigues; Verhemeldonck, Marc; Grauwen, Karolien; Dillen, Lieve; Mercken, Marc

doi:10.1016/j.chroma.2021.462299

Cited by 16 publications

(11 citation statements)

References 42 publications

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“…This antibody can recognize the microtubule-binding region of tau with high affinity for pThr217. JNJ-63733657 treatment has been shown to cause dose-dependent reductions of pTau in the CSF [ 83 ]. Now, JNJ-63733657 is in the phase II to evaluate its effect on cognitive decline in AD patients (NCT04619420) [ 84 ].…”

Section: Immunotherapies Based On Tau Proteinmentioning

confidence: 99%

Immunotherapy for Alzheimer’s disease: targeting β-amyloid and beyond

Song

Shi

Zhang

et al. 2022

Transl Neurodegener

114

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Alzheimer’s disease (AD) is the most common neurodegenerative disease in the elderly worldwide. However, the complexity of AD pathogenesis leads to discrepancies in the understanding of this disease, and may be the main reason for the failure of AD drug development. Fortunately, many ongoing preclinical and clinical studies will continually open up avenues to unravel disease mechanisms and guide strategies for AD diagnosis and drug development. For example, immunotherapeutic strategies targeting amyloid-β (Aβ) and tau proteins were once deemed almost certainly effective in clinical treatment due to the excellent preclinical results. However, the repeated failures of clinical trials on vaccines and humanized anti-Aβ and anti-tau monoclonal antibodies have resulted in doubts on this strategy. Recently, a new anti-Aβ monoclonal antibody (Aducanumab) has been approved by the US Food and Drug Administration, which brings us back to the realization that immunotherapy strategies targeting Aβ may be still promising. Meanwhile, immunotherapies based on other targets such as tau, microglia and gut-brain axis are also under development. Further research is still needed to clarify the forms and epitopes of targeted proteins to improve the accuracy and effectiveness of immunotherapeutic drugs. In this review, we focus on the immunotherapies based on Aβ, tau and microglia and their mechanisms of action in AD. In addition, we present up-to-date advances and future perspectives on immunotherapeutic strategies for AD.

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Section: Immunotherapies Based On Tau Proteinmentioning

confidence: 99%

Immunotherapy for Alzheimer’s disease: targeting β-amyloid and beyond

Song

Shi

Zhang

et al. 2022

Transl Neurodegener

114

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“…There was also a significant neuroinflammatory activation of astrocytes and microglia ( Jung et al, 2019 ). In addition, some studies have shown that magnetic resonance imaging (MRI)-based assessment of brain atrophy can be used to evaluate and stage Alzheimer’s disease, which laid a foundation for the specific neuroimaging changes in the brain of our study ( Bijttebier et al, 2021 ). Researchers have indicated that it is now possible to measure tau and amyloid beta (Aβ) protein in the brain.…”

Section: Introductionmentioning

confidence: 99%

Regional Homogeneity in Patients With Mild Cognitive Impairment: A Resting-State Functional Magnetic Resonance Imaging Study

Zhang

Liu

et al. 2022

Front. Aging Neurosci.

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ObjectiveTo analyze the potential changes in brain neural networks in resting state functional magnetic resonance imaging (rs-fMRI) scans by regional homogeneity (ReHo) in patients with mild cognitive impairment (MCI).MethodsWe recruited and selected 24 volunteers, including 12 patients (6 men and 6 women) with MCI and 12 healthy controls matched by age, sex, and lifestyle. All subjects were examined with rs-fMRI to evaluate changes in neural network connectivity, and the data were analyzed by ReHo method. Correlation analysis was used to investigate the relationship between ReHo values and clinical features in different brain regions of MCI patients. The severity of MCI was determined by the Mini-Mental State Examination (MMSE) scale.ResultsThe signals of the right cerebellum areas 4 and 5, left superior temporal, right superior temporal, left fusiform, and left orbital middle frontal gyri in the patient group were significantly higher than those in the normal group (P < 0.01 by t-test of paired samples). The signal intensity of the right inferior temporal and left inferior temporal gyri was significantly lower than that of the normal group (P < 0.01). The ReHO value for the left inferior temporal gyrus correlated negatively with disease duration, and the value for the right inferior temporal gyrus correlated positively with MMSE scores.ConclusionMild cognitive impairment in patients with pre- Alzheimer’s disease may be related to the excitation and inhibition of neural networks in these regions. This may have a certain guiding significance for clinical diagnosis.

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“…Moreover, tau phosphorylation at S1.2. T220 has never been reported before: 24 therefore, the singly phosphorylated TPSLPTPPTR measured with the abovementioned product ion was assigned to TPSLP(pT217)PPTR. Extracted ion chromatograms (XICs) of the targeted peptides measured in a solvent dilution of synthetic peptide standards and in a brain homogenate of transgenic mice dosed with Thiamet G are provided in Figure 2.…”

Section: Identification Of Tau O-glcnacylation Around S400 In Mouse B...mentioning

confidence: 93%

“…One explanation for the broader peak shapes of these Pro-rich peptides might be slow cis−trans peptide bond isomerization of Pro−Pro moieties during chromatographic separation, as observed by others. 24,31 Only a limited amount of information was gathered for localization of the O-GlcNAc site: one product ion with retention of the O-GlcNAc moiety was observed, representing y 8 fragmentation and thereby excluding O-GlcNAcylation at T181. Based on the available spectra, it was not possible to distinguish between O-GlcNAcylation at S184 or S185 (Figures S3.13−S3.15 and Table S3.5 in Supporting Information 3).…”

Section: Determination Of O-glcnac Sites In O-glcnacylated Recombinan...mentioning

confidence: 99%

“…The applied immunoprecipitation (IP) assay was based on a previously developed methodology. 24 Per IP reaction, 93.5 μL of Dynabeads protein G (Thermo Fisher Scientific) corresponding to 2.8 mg of beads was washed two times in LoBind Eppendorf tubes (Eppendorf) with 125 μL of 0.01% Tween 20 in PBS. The tubes were placed in a DynaMag-2 magnet (Thermo Fisher Scientific), and the supernatant was discarded.…”

Section: Sample Preparationmentioning

confidence: 99%

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IP-LC-MSMS Enables Identification of Three Tau O-GlcNAcylation Sites as O-GlcNAcase Inhibition Pharmacodynamic Readout in Transgenic Mice Overexpressing Human Tau

et al. 2023

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O-β-linked N-acetylglucosaminylation (O-GlcNAcylation) modulates tau phosphorylation and aggregation: the pharmacological increase of tau O-GlcNAcylation upon treatment with inhibitors of O-GlcNAc hydrolase (OGA) constitutes a potential strategy to tackle neurodegenerative diseases. Analysis of tau O-GlcNAcylation could potentially be used as a pharmacodynamic biomarker both in preclinical and clinical studies. The goal of the current study was to confirm tau O-GlcNAcylation at S400 as a pharmacodynamic readout of OGA inhibition in P301S transgenic mice overexpressing human tau and treated with the OGA inhibitor Thiamet G and to explore if additional O-GlcNAcylation sites on tau could be identified. As a first step, an immunoprecipitation-liquid chromatography-mass spectrometry (IP-LC-MS) methodology was developed to monitor changes in O-GlcNAcylation around S400 of tau in mouse brain homogenate (BH) extracts. Second, additional O-GlcNAc sites were identified in in-house produced recombinant O-GlcNAcylated human tau at relatively high concentrations, thereby facilitating collection of informative LC-MS data for identification of low-concentration O-GlcNAc-tryptic tau peptides in human transgenic mouse BH extracts. This strategy enabled, for the first time, identification of three low abundant N-terminal and mid-domain O-GlcNAc sites of tau (at S208, S191, and S184 or S185) in human transgenic mouse BH. Data are openly available at (doi: 10.17632/jp57yk9469.1; doi: 10.17632/8n5j45dnd8.1; doi: 10.17632/h5vdrx4n3d.1).

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Development of immunoprecipitation – two-dimensional liquid chromatography – mass spectrometry methodology as biomarker read-out to quantify phosphorylated tau in cerebrospinal fluid from Alzheimer disease patients

Cited by 16 publications

References 42 publications

Immunotherapy for Alzheimer’s disease: targeting β-amyloid and beyond

Immunotherapy for Alzheimer’s disease: targeting β-amyloid and beyond

Regional Homogeneity in Patients With Mild Cognitive Impairment: A Resting-State Functional Magnetic Resonance Imaging Study

IP-LC-MSMS Enables Identification of Three Tau O-GlcNAcylation Sites as O-GlcNAcase Inhibition Pharmacodynamic Readout in Transgenic Mice Overexpressing Human Tau

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