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. 2023 Sep;94(9):726-737.
doi: 10.1136/jnnp-2022-330796. Epub 2023 Apr 19.

Granulocyte activation markers in cerebrospinal fluid differentiate acute neuromyelitis spectrum disorder from multiple sclerosis

David Leppert  1 , Mitsuru Watanabe  2 , Sabine Schaedelin  3 , Fredrik Piehl  4 , Roberto Furlan  5 , Matteo Gastaldi  6 , Jeremy Lambert  7 , Björn Evertsson  4 , Katharina Fink  4 , Takuya Matsushita  2 , Katsuhisa Masaki  2 , Noriko Isobe  2 , Jun-Ichi Kira  2   8   9 , Pascal Benkert  3 , Aleksandra Maceski  10 , Eline Willemse  11 , Johanna Oechtering  11 , Annette Orleth  11 , Stephanie Meier  11 , Jens Kuhle  11
Affiliations

Granulocyte activation markers in cerebrospinal fluid differentiate acute neuromyelitis spectrum disorder from multiple sclerosis

David Leppert et al. J Neurol Neurosurg Psychiatry. 2023 Sep.

Abstract

Background: Granulocyte invasion into the brain is a pathoanatomical feature differentiating neuromyelitis optica spectrum disorder (NMOSD) from multiple sclerosis (MS). We aimed to determine whether granulocyte activation markers (GAM) in cerebrospinal fluid (CSF) can be used as a biomarker to distinguish NMOSD from MS, and whether levels associate with neurological impairment.

Methods: We quantified CSF levels of five GAM (neutrophil elastase, myeloperoxidase, neutrophil gelatinase-associated lipocalin, matrixmetalloproteinase-8, tissue inhibitor of metalloproteinase-1), as well as a set of inflammatory and tissue-destruction markers, known to be upregulated in NMOSD and MS (neurofilament light chain, glial fibrillary acidic protein, S100B, matrix metalloproteinase-9, intercellular adhesion molecule-1, vascular cellular adhesion molecule-1), in two cohorts of patients with mixed NMOSD and relapsing-remitting multiple sclerosis (RRMS).

Results: In acute NMOSD, GAM and adhesion molecules, but not the other markers, were higher than in RRMS and correlated with actual clinical disability scores. Peak GAM levels occurred at the onset of NMOSD attacks, while they were stably low in MS, allowing to differentiate the two diseases for ≤21 days from onset of clinical exacerbation. Composites of GAM provided area under the curve values of 0.90-0.98 (specificity of 0.76-1.0, sensitivity of 0.87-1.0) to differentiate NMOSD from MS, including all anti-aquaporin-4 protein (aAQP4)-antibody-negative patients who were untreated.

Conclusions: GAM composites represent a novel biomarker to reliably differentiate NMOSD from MS, including in aAQP4- NMOSD. The association of GAM with the degree of concurrent neurological impairment provides evidence for their pathogenic role, in turn suggesting them as potential drug targets in acute NMOSD.

Keywords: CSF; clinical neurology; molecular biology; multiple sclerosis; neuroimmunology.

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Conflict of interest statement

Competing interests: DL is Chief Medical Officer of GeNeuro. MW received speaker honoraria from Novartis Pharma, Chugai Pharmaceutical, Biogen Japan and Alexion. FP has received research grants from Janssen, Merck KGaA and UCB, and fees for serving on DMC in clinical trials with Chugai, Lundbeck and Roche, and preparation of witness report for Novartis. RF has received speaker fees for teaching and workshops from Biogen, Merck, Novartis, Roche, Teva and Alexion. For educational activities, courses or research, he has received unrestricted grants from Biogen, EMD Serono. JL is an employee of Quanterix. BE has received travel grants for ECTRIMS 2018 from Roche. KF has served on advisory boards and received speaker honoraria from Biogen, Roche and Merck and received research funds from Amicus. TM received speaker honoraria from Biogen Japan, Chugai Pharmaceutical, Alexion Pharmaceuticals, Novartis Pharma and Takeda Pharmaceutical. KM received speaker honoraria from Novartis Pharma, Chugai Pharmaceutical and Nihon Pharmaceutical. NI received grant support from Mitsubishi Tanabe Pharma, Osoegawa Neurology Clinic, Bayer Yakuhin and Japan Blood Products Organization and speaker honoraria from Novartis Pharma, Biogen Japan, Alexion, Mitsubishi Tanabe Pharma, Chugai Pharmaceutical, Teijin Pharma and Eisai. J-IK received research funds from Dainippon Sumitomo Pharma, Daiichi Sankyo, Mitsubishi Tanabe Pharma and Kyowa Kensetsukougyo, and consultancy fees, speaking fees and/or honoraria from Novartis Pharma, Mitsubishi Tanabe Pharma, CSL Behring, Biogen Japan, Teijin Health Care, the Takeda Pharmaceutical, Kyowa Kirin, Ono Pharmaceutical, Alexion Pharmaceuticals, Tsumura, Ricoh, EMC and Eisai. JO served on advisory boards for Roche and Merck. JK received speaker fees, research support, travel support and/or served on advisory boards by the Progressive MS Alliance, Swiss MS Society, Swiss National Research Foundation (320030_189140/1), University of Basel, Biogen, Celgene, Merck, Novartis, Octave Bioscience, Roche, Sanofi. No other disclosures were reported.

Figures

Figure 1
Figure 1
Association between clinical disease severity and granulocyte activation markers levels in patients with NMOSD. Biomarker values are in pg/mL. Values on x-axis show the EDSS score at the time point of lumbar puncture. Regression lines show correlations in NMOSD patients without (acute formula image; s/c:formula image) corticosteroid pre-treatment; patients with (acute: formula image; s/c:formula image) corticosteroid pre-treatment. Open symbols designate aAQP4− patients. The Spearman’s correlation analysis of all, acute and s/c cohorts of patients showed significant correlations with the EDSS score; note that intercellular adhesion molecule-1 and vascular cellular adhesion molecule-1 levels correlated as well with EDSS scores (see online supplemental table 3). EDSS, Expanded Disability Status Scale; MMP-8, matrix metalloproteinase 8; MPO, myeloperoxidase; nEla, neutrophil elastase; NGAL, neutrophil gelatinase-associated lipocalin; NMOSD, neuromyelitis optica spectrum disorder; TIMP-1, tissue inhibitor of metalloproteinase-1.
Figure 2
Figure 2
Modelled kinetics of biomarker levels in NMOSD and RRMS in function of days after disease exacerbation. Biomarker values are in pg/mL. Values on x-axis show days after disease exacerbations. Dotted lines determine 95% CI, based on all patients. (A) Pattern 1: increased in NMOSD, stably low (nEla, MPO. MMP-8, GFAP, S100B, ICAM-1, VCAM-1), or slightly increasing over time (NGAL, TIMP-1) in RRMS. (B) Pattern 2: increased in both NMOSD and RRMS at disease exacerbation: MMP-9 and CXCL13; pattern 3: stably high in NMOSD and RRMS: NfL. NMOSD: yellow (pooled cohorts), red (discovery cohort only); RRMS: brown (pooled cohorts), green (discovery cohort only). CXCL13, C-X-C motif chemokine 13; GFAP, glial fibrillar acidic protein; ICAM-1, intercellular adhesion molecule-1; MMP, matrix metalloproteinase; MPO, myeloperoxidase; nEla, neutrophil elastase; NGAL, neutrophil gelatinase-associated lipocalin; NfL, neurofilament light chain; NMOSD, neuromyelitis optica spectrum disorder; RRMS, relapsing-remitting multiple sclerosis; TIMP-1, tissue inhibitor of metalloproteinase-1; S100B, S100 calcium-binding protein; VCAM-1, vascular cell adhesion molecule-1.
Figure 3
Figure 3
ROC curves for the differentiation between NMOSD and RRMS in patients without corticosteroid pretreatment without (A, B) and with (C, D) time as covariate A B C D ROC curves of individual (A, C) GAM and their composites (B, D) (composite 1=nEla+ MPO+NGAL+MMP-8; composite 2=nEla+MPO+NGAL+MMP-8+TIMP-1). For numerical values of AUC (95% CIs), specificity and sensitivity, see table 4. AUC, area under the curve; MMP-8, matrix metalloproteinase 8; MPO, myeloperoxidase; nEla, neutrophil elastase; NGAL, neutrophil gelatinase-associated lipocalin; NMOSD, neuromyelitis optica spectrum disorder; ROC, receiver operating characteristics; RRMS, relapsing-remitting multiple sclerosis; TIMP-1, tissue inhibitor of metalloproteinase-1.
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References

    1. Kuchling J, Paul F. Visualizing the central nervous system: imaging tools for multiple sclerosis and neuromyelitis optica spectrum disorders. Front Neurol 2020;11(June):450. 10.3389/fneur.2020.00450 - DOI - PMC - PubMed
    1. Hamid SHM, Whittam D, Mutch K, et al. . What proportion of AQP4-igG-negative NMO spectrum disorder patients are MOG-igg positive? A cross sectional study of 132 patients. J Neurol 2017;264:2088–94. 10.1007/s00415-017-8596-7 - DOI - PMC - PubMed
    1. Jarius S, Paul F, Franciotta D, et al. . Cerebrospinal fluid findings in aquaporin-4 antibody positive neuromyelitis optica: results from 211 lumbar punctures. J Neurol Sci 2011;306:82–90. 10.1016/j.jns.2011.03.038 - DOI - PubMed
    1. Schmetzer O, Lakin E, Roediger B, et al. . Anti-aquaporin 4 igG is not associated with any clinical disease characteristics in neuromyelitis optica spectrum disorder. Front Neurol 2021;12(March):635419. 10.3389/fneur.2021.635419 - DOI - PMC - PubMed
    1. Juryńczyk M, Weinshenker B, Akman-Demir G, et al. . Status of diagnostic approaches to AQP4-igG seronegative NMO and NMO/MS overlap syndromes. J Neurol 2016;263:140–9. 10.1007/s00415-015-7952-8 - DOI - PMC - PubMed

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