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Exploring Cerebrospinal Fluid IgG N-Glycosylation as Potential Biomarker for Amyotrophic Lateral Sclerosis

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Abstract

Amyotrophic lateral sclerosis (ALS) is a fatal motor neuron disease for which the existing candidate biomarkers (neurofilaments) have low specificity. Changes in blood IgG N-glycosylation have been observed in several diseases, including ALS, whereas cerebrospinal fluid (CSF) IgG has been less studied. Here, we characterized N-glycans of CSF IgG from ALS patients in comparison with a control group of other neurological diseases. Cerebrospinal fluid was collected from patients with ALS (n = 26) and other neurological diseases (n = 10). N-Glycans were released from CSF purified IgG with peptide N-glycosidase F, labeled with 2-aminobenzamide and analyzed by NP-HPLC chromatography in combination with exoglycosidase digestion and MALDI-TOF mass spectrometry. The N-glycosylation profile of ALS CSF IgG consisted of diantennary N-glycans predominantly with proximal fucose and some bisecting GlcNAc; agalacto-, mono-, and digalactosylated as well as α2,6-sialylated structures were detected. Differences between ALS and control patients were observed; most relevant was the increase in ALS CSF IgG of the level of galactosylated structures defined here as Gal-index (median 46.87 and 40.50% for ALS and controls, respectively; p = 0.006). The predictive value of the Gal-index (AUC = 0.792, p = 0.007) considering ROC analysis had potential utility as a diagnostic test for ALS and was comparable to that of phosphoneurofilament heavy chain (AUC = 0.777, p = 0.011), which was used as benchmark marker for our group of patients. The results provide the basis to further explore the potential of IgG N-glycan galactosylation as biomarker for ALS by using larger cohorts of patients and controls.

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Funding

This work was supported by the EU JPND project SOPHIA (JPND/0003/2011), Fundação para a Ciência e a Tecnologia (FCT); Portugal and Euronanomed 2 ERA-NET project GlioEx (ENMed/0001/2013), FCT, Portugal; iNOVA4Health Research Unit (LISBOA-01-0145-FEDER-007344), which is cofunded by FCT/Ministério da Ciência e do Ensino Superior, through national funds; and by FEDER under the PT2020 Partnership Agreement.

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Authors and Affiliations

  1. Laboratory of Glycobiology, Instituto de Tecnologia Química e Biológica António Xavier, Universidade Nova de Lisboa, Avenida da República, 2780-157, Oeiras, Portugal

    Julia Costa

  2. GlycoThera GmbH, Feodor-Lynen Strasse 35, 30625, Hannover, Germany

    Linda Streich & Harald S. Conradt

  3. Institute of Physiology, Instituto de Medicina Molecular-Faculty of Medicine, University of Lisbon, Lisbon, Portugal

    Susana Pinto, Ana Pronto-Laborinho & Mamede de Carvalho

  4. Helmholtz-Zentrum für Infektionsforschung, 38124, Braunschweig, Germany

    Manfred Nimtz

  5. Department Neurosciences and Mental Health, Hospital de Santa Maria-CHLN, Lisbon, Portugal

    Mamede de Carvalho

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  1. Julia Costa
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  2. Linda Streich
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  4. Ana Pronto-Laborinho
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Correspondence to Julia Costa.

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Patients signed permission for biobank storage, and further studies were agreed by the local Ethic’s committee. The research was done in accordance with the Helsinki Declaration as revised in 2013 (www.wma.net/policies-post/wma-declaration-of-helsinki-ethical-principles-for-medical-research-involving-human-subjects).

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Costa, J., Streich, L., Pinto, S. et al. Exploring Cerebrospinal Fluid IgG N-Glycosylation as Potential Biomarker for Amyotrophic Lateral Sclerosis. Mol Neurobiol 56, 5729–5739 (2019). https://doi.org/10.1007/s12035-019-1482-9

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