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Meta-Analysis
. 2024 Jan 8;25(2):787.
doi: 10.3390/ijms25020787.

The Use of Nitrosative Stress Molecules as Potential Diagnostic Biomarkers in Multiple Sclerosis

Saskia Räuber  1 , Moritz Förster  1   2 , Julia Schüller  1 , Alice Willison  1 , Kristin S Golombeck  1 , Christina B Schroeter  1 , Menekse Oeztuerk  1 , Robin Jansen  1 , Niklas Huntemann  1 , Christopher Nelke  1 , Melanie Korsen  1 , Katinka Fischer  1 , Ruth Kerkhoff  1   2 , Yana Leven  1 , Patricia Kirschner  1 , Tristan Kölsche  1 , Petyo Nikolov  1 , Mohammed Mehsin  1 , Gelenar Marae  1 , Alma Kokott  1 , Duygu Pul  1 , Julius Schulten  1 , Niklas Vogel  1 , Jens Ingwersen  1 , Tobias Ruck  1 , Marc Pawlitzki  1 , Sven G Meuth  1 , Nico Melzer  1 , David Kremer  1   3
Affiliations
Meta-Analysis

The Use of Nitrosative Stress Molecules as Potential Diagnostic Biomarkers in Multiple Sclerosis

Saskia Räuber et al. Int J Mol Sci. .

Abstract

Multiple sclerosis (MS) is an autoimmune disease of the central nervous system (CNS) of still unclear etiology. In recent years, the search for biomarkers facilitating its diagnosis, prognosis, therapy response, and other parameters has gained increasing attention. In this regard, in a previous meta-analysis comprising 22 studies, we found that MS is associated with higher nitrite/nitrate (NOx) levels in the cerebrospinal fluid (CSF) compared to patients with non-inflammatory other neurological diseases (NIOND). However, many of the included studies did not distinguish between the different clinical subtypes of MS, included pre-treated patients, and inclusion criteria varied. As a follow-up to our meta-analysis, we therefore aimed to analyze the serum and CSF NOx levels in clinically well-defined cohorts of treatment-naïve MS patients compared to patients with somatic symptom disorder. To this end, we analyzed the serum and CSF levels of NOx in 117 patients (71 relapsing-remitting (RR) MS, 16 primary progressive (PP) MS, and 30 somatic symptom disorder). We found that RRMS and PPMS patients had higher serum NOx levels compared to somatic symptom disorder patients. This difference remained significant in the subgroup of MRZ-negative RRMS patients. In conclusion, the measurement of NOx in the serum might indeed be a valuable tool in supporting MS diagnosis.

Keywords: biomarkers; nitrosative stress molecules; primary progressive multiple sclerosis; relapsing remitting multiple sclerosis.

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Conflict of interest statement

S.R. received travel grants from Merck Healthcare Germany GmbH, Alexion Pharmaceuticals, Bristol Myers Squibb, the German Neurological Society, and the American Academy of Neurology. She served on a scientific advisory board from Merck Healthcare Germany GmbH and received honoraria for lecturing from Roche. Her research was supported by Novartis, ‘Stiftung zur Förderung junger Neurowissenschaftler’ and ‘Else Kröner-Fresenius-Stiftung’. M.F. received travel grants from Merck Healthcare Germany GmbH, Novartis, the German Neurological Society, and benefitted in kind from Biogen. He served on a scientific advisory board from Swedish Orphan Biovitrum GmbH. C.B.S. received travel grants from Alexion Pharmaceuticals. She served on a scientific advisory board from Merck Healthcare Germany GmbH. N.H. received honoraria for lecturing meetings from Argenx and travel reimbursements and meeting attendance fees from Alexion, Merck, and Novartis. He served on a scientific advisory board from Viatris. M.P. received honoraria for lecturing and travel expenses for attending meetings from Alexion, ArgenX, Bayer Health Care, Biogen, Hexal, Merck Serono, Novartis, Roche, Sanofi-Aventis, and Teva. His research is funded by Biogen, Hexal, Merck Serono, Novartis, and Teva. S.G.M. received honoraria for lecturing and travel expenses for attending meetings from Almirall, Amicus Therapeutics Germany, Bayer HealthCare, Biogen, Celgene, Diamed, Genzyme, MedDay Pharmaceuticals, Merck Serono, Novartis, Novo Nordisk, ONO Pharma, Roche, Sanofi-Aventis, Chugai Pharma, QuintilesIMS, and Teva. His research is funded by the German Ministry for Education and Research (BMBF), Bundesinstitut für Risikobewertung (BfR), Deutsche Forschungsgemeinschaft (DFG), Else Kröner Fresenius Foundation, Gemeinsamer Bundesausschuss (G-BA), German Academic Exchange Service, Hertie Foundation, Interdisciplinary Center for Clinical Studies (IZKF) Muenster, German Foundation Neurology and by Alexion, Almirall, Amicus Therapeutics Germany, Biogen, Diamed, Fresenius Medical Care, Genzyme, HERZ Burgdorf, Merck Serono, Novartis, ONO Pharma, Roche, and Teva. N.M. has received honoraria for lecturing and travel expenses for attending meetings from Biogen Idec, GlaxoSmith Kline, Teva, Novartis Pharma, Bayer Healthcare, Genzyme, Alexion Pharmaceuticals, Fresenius Medical Care, Diamed, UCB Pharma, AngeliniPharma, BIAL and Sanofi-Aventis, has received royalties for consulting from UCB Pharma, Alexion Pharmaceuticals, and Sanofi-Aventis, and has received financial research support from Euroimmun, Fresenius Medical Care, Diamed, Alexion Pharmaceuticals, and Novartis Pharma. D.K. received travel grants from GeNeuro and Merck, refund of congress participation fees from GeNeuro, Merck and Servier, consulting fees from Grifols, payment for lectures from Grifols, support for research projects from Teva, and was funded by the Deutsche Forschungsgemeinschaft (DFG). The remaining authors declare no conflicts of interest.

Figures

Figure 1
Figure 1
RRMS patients feature elevated NOx serum levels compared to somatic symptom disorder patients (Soma). (A–E) Violin plots with overlaying box plots comparing two groups. Boxes display the median as well as the 25th and 75th percentiles. The whiskers extend from the hinge to the largest and smallest values, respectively, but no further than 1.5 * IQR from the hinge. (F,G) Simple linear regression using either age or EDSS as independent and NOx serum as dependent variable. The area in-between the dotted lines shows the 95% confidence band. (H,I) Simple linear regression using NOx serum as independent and either QAlbumin or NOx CSF as dependent variable. The area in-between the dotted lines shows the 95% confidence band. (J) Pie charts comparing the percentage of patients with acute relapse and active lesions on cMRI at time of sampling between the high and low serum NOx group. CSF—Cerebrospinal fluid, EDSS—Expanded Disability Status Scale, IQR—Interquartile range, M—Months, MRI—Magnetic resonance imaging, MRZ—Antibody indices (AI) against measles, rubella, and varicella zoster virus. MRZ was defined ‘positive’ if at least two out of three AI were higher than 1.5, neg—Negative, NOx—Nitrosative stress molecules, ns—Not significant, OCBs—Oligoclonal bands, Q—CSF/serum ratio, RRMS—Relapsing–remitting Multiple Sclerosis, Soma—Somatic symptom disorder, *—multiplied by.
Figure 2
Figure 2
CSF NOx levels do not differ significantly between RRMS and Soma patients. (A,B) Violin plots with overlaying box plots comparing two groups. Boxes display the median as well as the 25th and 75th percentiles. The whiskers extend from the hinge to the largest and smallest values, respectively, but no further than 1.5 * IQR from the hinge. (C,D) Simple linear regression using either age or EDSS as independent and NOx CSF as dependent variable. The area in-between the dotted lines shows the 95% confidence band. (E) Simple linear regression using NOx CSF as independent and QAlbumin as dependent variable. The area in-between the dotted lines shows the 95% confidence band. BL—Baseline, CSF—Cerebrospinal fluid, EDSS—Expanded Disability Status Scale, IQR—Interquartile range, NOx—Nitrosative stress molecules, ns—Not significant, Q—CSF/serum ratio, RRMS—Relapsing–remitting Multiple Sclerosis, Soma—Somatic symptom disorder.
Figure 3
Figure 3
Increased nitrosative stress in serum of PPMS patients compared to Soma patients. (A–C) Violin plots with overlaying box plots comparing two groups. Boxes display the median as well as the 25th and 75th percentiles. The whiskers extend from the hinge to the largest and smallest values, respectively, but no further than 1.5 * IQR from the hinge. (D) Simple linear regression using age as independent and NOx serum as dependent variable. The area in-between the dotted lines shows the 95% confidence band. IQR—Interquartile range, NOx—Nitrosative stress molecules, ns—Not significant, PPMS—Primary progressive Multiple Sclerosis, Q—CSF/serum ratio, RRMS—Relapsing–remitting Multiple Sclerosis, Soma—Somatic symptom disorder, *—multiplied by.
Figure 4
Figure 4
Study design. CSF—Cerebrospinal fluid, NOx—Nitrosative stress molecules, PPMS—Primary progressive Multiple Sclerosis, RRMS—Relapsing–remitting Multiple Sclerosis, Soma—Somatic symptom disorder. Parts of the figure were drawn by using pictures from Servier Medical Art (https://smart.servier.com/, accessed on 1 September 2023). Servier Medical Art by Servier is licensed under a Creative Commons Attribution 3.0 Unported License (https://creativecommons.org/licenses/by/3.0/, accessed on 1 September 2023).
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