Proteomic predictors of physical, cognitive and imaging outcomes in multiple sclerosis: 5-year follow-up study

Background: A quantitative measurement of serum proteome biomarkers that would associate with disease progression endpoints can provide risk stratification for persons with multiple sclerosis and supplement the clinical decision-making process. Materials and Methods: 202 persons with multiple sclerosis were enrolled in a longitudinal study with measurements at two time points with an average follow-up time of 5.4 years. Clinical measures included the Expanded Disability Status Scale, Timed 25-foot Walk, 9-Hole Peg and Symbol Digit Modalities Tests. Subjects underwent magnetic resonance imaging to determine the volumetric measures of the whole brain, gray matter, deep gray matter and lateral ventricles. Serum samples were analyzed using a custom immunoassay panel on the Olink platform and concentrations of 18 protein biomarkers were measured. Linear mixed-effects models and adjustment for multiple comparisons were performed. Results: Subjects had a significant 55.6% increase in Chemokine Ligand 20 (9.7pg/mL vs. 15.1pg/mL, p<0.001) and Neurofilament light polypeptide (10.5 pg/ml vs. 11.5 pg/ml, p=0.003) at the follow-up time point. Additional changes in CUB domain-containing protein 1, Contactin 2, Glial fibrillary acidic protein, Myelin oligodendrocyte glycoprotein, and Osteopontin were noted but did not survive multiple comparisons correction. Worse clinical performance in the 9-HPT was associated with Neurofilament light polypeptide (p=0.001). Increases in several biomarker candidates were correlated with greater neurodegenerative changes as measured by different brain volumes. Conclusion: Multiple proteins representing diverse biological pathways (neuroinflammation, immune modulation, and neuroaxonal integrity) associate with physical, cognitive and radiographic outcomes. Future studies should determine the utility of multiple protein assays in routine clinical care. ### Competing Interest Statement Kian Jalaleddini, Ferhan Qureshi, Anisha Keshavan Shannon McCurdy, Kelly Leyden, and Ati Ghoreyshi are employees of and either hold stock or stock options at Octave Bioscience. Dejan Jakimovski received honoraria for serving on the advisory board of AstraZeneca. He also serves as an Associate Editor for Clinical Neurology and Neurosurgery and compensated by Elsevier B.V. Niels Bergsland has nothing to disclose. Murali Ramanathan received research funding from the National Multiple Sclerosis Society, Department of Defense and National Institute of Neurological Diseases and Stroke. Michael G. Dwyer received compensation from Keystone Heart for consultant fees. He received financial support for research activities from Bristol Myers Squibb, Mapi Pharma, Keystone Heart, Protembis and V-WAVE Medical. Bianca Weinstock-Guttman received honoraria for serving in advisory boards and educational programs from Biogen Idec, Novartis, Genentech, Genzyme and Sanofi, Janssen, Abbvie and Bayer. She also received support for research activities from the National Institutes of Health, National Multiple Sclerosis Society, Department of Defense, and Biogen Idec, Novartis, Genentech, Genzyme and Sanofi. Dr. Robert Zivadinov has received personal compensation for speaking and consulting activities with Bristol Myers Squibb, Sanofi, Novartis, EMD Serono, Janssen, Filterlex, 415 Capital, and Sana Biotechnologies. Dr. Zivadinov has received research support from the U.S. National Institutes of Health, the U.S. National Science Foundation, the U.S. Department of Defense, the U.S. National Multiple Sclerosis Society, EMD Serono, Novartis, Mapi Pharma, Protembis, Octave Biosciences, CorEvitas, and V-WAVE Medical. Dr. Zivadinov serves on the editorial board of J Alzheimers Dis, BMC Med, BMC Neurol, Veins and Lymphatics, Front, Neurol, Trans Neurosci, and Clinical CNS Drugs. ### Funding Statement This study did not receive any funding ### Author Declarations I confirm all relevant ethical guidelines have been followed, and any necessary IRB and/or ethics committee approvals have been obtained. Yes The details of the IRB/oversight body that provided approval or exemption for the research described are given below: University at Buffalo Institutional Review Board gave ethical approval for this work I confirm that all necessary patient/participant consent has been obtained and the appropriate institutional forms have been archived, and that any patient/participant/sample identifiers included were not known to anyone (e.g., hospital staff, patients or participants themselves) outside the research group so cannot be used to identify individuals. Yes I understand that all clinical trials and any other prospective interventional studies must be registered with an ICMJE-approved registry, such as ClinicalTrials.gov. I confirm that any such study reported in the manuscript has been registered and the trial registration ID is provided (note: if posting a prospective study registered retrospectively, please provide a statement in the trial ID field explaining why the study was not registered in advance). Yes I have followed all appropriate research reporting guidelines, such as any relevant EQUATOR Network research reporting checklist(s) and other pertinent material, if applicable. Yes All data produced in the present work are contained in the manuscript and supplementary materials.