{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,28]],"date-time":"2026-10-28T19:08:32Z","timestamp":1793214512848,"version":"4.5.0"},"reference-count":44,"publisher":"MDPI AG","issue":"11","license":[{"start":{"date-parts":[[2025,11,1]],"date-time":"2025-11-01T00:00:00Z","timestamp":1761955200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Genes"],"abstract":"<jats:p>Background: Multiple Sclerosis (MS) is a chronic, autoimmune, multifactorial, and complex disorder of the central nervous system (CNS), affecting more than 2 million individuals globally. Genome-wide association studies (GWAS) have explained only a small fraction of its high heritability, highlighting the need for alternative approaches to identify rare genetic variants that contribute to its etiology. To address this, we performed whole-exome sequencing (WES) in a multi-affected family. Methods: WES was performed in a MS multigenerational family comprising two affected sisters, their two healthy brothers, and one affected son. Results: Bioinformatics analysis identified 47 co-segregating rare variants. Three missense variants in genes involved in inflammation, autoimmunity, and demyelinization were identified as the most promising candidates: c.443 C&gt;T, p.Pro148Leu in the RTN4 gene, c.1678 T&gt;G, p.Phe560Val in the JAK2 gene, and c.3449 A&gt;G, p.Tyr1150Cys in the DUOX2 gene. Protein modeling and in silico tools suggest that the three selected variants may have a significant impact on protein function. Conclusions: We identified novel candidate genes for MS in a multiplex family, providing evidence for an oligogenic model of disease susceptibility. Further replication and functional studies are required to validate these preliminary results.<\/jats:p>","DOI":"10.3390\/genes16111311","type":"journal-article","created":{"date-parts":[[2025,11,3]],"date-time":"2025-11-03T13:22:36Z","timestamp":1762176156000},"page":"1311","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Exome Sequencing Uncovers Genetic Drivers of Multiple Sclerosis in a Multiplex Family"],"prefix":"10.3390","volume":"16","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-1209-8554","authenticated-orcid":false,"given":"Carla","family":"Lintas","sequence":"first","affiliation":[{"name":"Research Unit of Medical Genetics, Department of Medicine, University Campus-Biomedico of Rome, 00128 Roma, Italy"},{"name":"Operative Research Unit of Medical Genetics, Fondazione Policlinico Universitario Campus Bio-Medico, 00128 Roma, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0001-2620-1252","authenticated-orcid":false,"given":"Simone","family":"Bonora","sequence":"additional","affiliation":[{"name":"Department of Chemistry and Biology \u201cA. Zambelli\u201d, University of Salerno, 84084 Fisciano, SA, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9510-5205","authenticated-orcid":false,"given":"Anna","family":"Marabotti","sequence":"additional","affiliation":[{"name":"Department of Chemistry and Biology \u201cA. Zambelli\u201d, University of Salerno, 84084 Fisciano, SA, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5001-285X","authenticated-orcid":false,"given":"Claudio","family":"Tabolacci","sequence":"additional","affiliation":[{"name":"Coordination and Promotion of Research, Istituto Superiore di Sanit\u00e0, Viale Regina Elena 299, 00161 Rome, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6659-0280","authenticated-orcid":false,"given":"Maria Luisa","family":"Scattoni","sequence":"additional","affiliation":[{"name":"Coordination and Promotion of Research, Istituto Superiore di Sanit\u00e0, Viale Regina Elena 299, 00161 Rome, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fioravante","family":"Capone","sequence":"additional","affiliation":[{"name":"Operative Research Unit of Neurology, Fondazione Policlinico Universitario Campus Bio-Medico, 00128 Roma, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mariagrazia","family":"Rossi","sequence":"additional","affiliation":[{"name":"Operative Research Unit of Neurology, Fondazione Policlinico Universitario Campus Bio-Medico, 00128 Roma, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9113-5925","authenticated-orcid":false,"given":"Vincenzo","family":"Di Lazzaro","sequence":"additional","affiliation":[{"name":"Operative Research Unit of Neurology, Fondazione Policlinico Universitario Campus Bio-Medico, 00128 Roma, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6775-5972","authenticated-orcid":false,"given":"Fiorella","family":"Gurrieri","sequence":"additional","affiliation":[{"name":"Research Unit of Medical Genetics, Department of Medicine, University Campus-Biomedico of Rome, 00128 Roma, Italy"},{"name":"Operative Research Unit of Medical Genetics, Fondazione Policlinico Universitario Campus Bio-Medico, 00128 Roma, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,11,1]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"21","DOI":"10.1007\/s44197-025-00353-6","article-title":"Epidemiology of Multiple Sclerosis: Global, Regional, National and Sub-National-Level Estimates and Future Projections","volume":"15","author":"Khan","year":"2025","journal-title":"J. Epidemiol. Glob. Health"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"693","DOI":"10.1001\/jamaneurol.2023.1135","article-title":"Population-Based Estimates for the Prevalence of Multiple Sclerosis in the United States by Race, Ethnicity, Age, Sex, and Geographic Region","volume":"80","author":"Hittle","year":"2023","journal-title":"JAMA Neurol."},{"key":"ref_3","doi-asserted-by":"crossref","unstructured":"Clarelli, F., Barizzone, N., Mangano, E., Zuccal\u00e0, M., Basagni, C., Anand, S., Sorosina, M., Mascia, E., Santoro, S. (2022). Contribution of Rare and Low-Frequency Variants to Multiple Sclerosis Susceptibility in the Italian Continental Population. Front. Genet., 12.","DOI":"10.3389\/fgene.2021.800262"},{"key":"ref_4","unstructured":"International Multiple Sclerosis Genetics Consortium (2018). Low-Frequency and Rare-Coding Variation Contributes to Multiple Sclerosis Risk. Cell, 175, 1679\u20131687.e7."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"641","DOI":"10.1016\/j.bbi.2010.01.014","article-title":"Infiltration of Th1 and Th17 cells and activation of microglia in the CNS during the course of experimental autoimmune encephalomyelitis","volume":"24","author":"Murphy","year":"2010","journal-title":"Brain Behav. Immun."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"1142","DOI":"10.4049\/jimmunol.1900311","article-title":"IL-11 Induces Encephalitogenic Th17 Cells in Multiple Sclerosis and Experimental Autoimmune Encephalomyelitis","volume":"203","author":"Zhang","year":"2019","journal-title":"J. Immunol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"e2221007120","DOI":"10.1073\/pnas.2221007120","article-title":"IL-11 induces NLRP3 inflammasome activation in monocytes and inflammatory cell migration to the central nervous system","volume":"120","author":"Seyedsadr","year":"2023","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_8","doi-asserted-by":"crossref","unstructured":"Everest, E., Ahangari, M., Uygunoglu, U., Tutuncu, M., Bulbul, A., Saip, S., Duman, T., Sezerman, U., Reich, D.S., and Riley, B.P. (2022). Investigating the role of common and rare variants in multiplex multiple sclerosis families reveals an increased burden of common risk variation. Sci. Rep., 12.","DOI":"10.1038\/s41598-022-21484-x"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"e1018","DOI":"10.1002\/cti2.1018","article-title":"Genome-wide association studies of multiple sclerosis","volume":"7","author":"Cotsapas","year":"2018","journal-title":"Clin. Transl. Immunol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"a028944","DOI":"10.1101\/cshperspect.a028944","article-title":"Lifestyle and Environmental Factors in Multiple Sclerosis","volume":"9","author":"Alfredsson","year":"2019","journal-title":"Cold Spring Harb. Perspect. Med."},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"Goecks, J., Nekrutenko, A., Taylor, J., and Galaxy Team (2010). Galaxy: A comprehensive approach for supporting accessible, reproducible, and transparent computational research in the life sciences. Genome Biol., 11.","DOI":"10.1186\/gb-2010-11-8-r86"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"1754","DOI":"10.1093\/bioinformatics\/btp324","article-title":"Fast and accurate short read alignment with Burrows-Wheeler transform","volume":"25","author":"Li","year":"2009","journal-title":"Bioinformatics"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"405","DOI":"10.1038\/gim.2015.30","article-title":"Standards and guidelines for the interpretation of sequence variants: A joint consensus recommendation of the American College of Medical Genetics and Genomics and the Association for Molecular Pathology","volume":"17","author":"Richards","year":"2015","journal-title":"Genet. Med."},{"key":"ref_14","doi-asserted-by":"crossref","unstructured":"Stelzer, G., Plaschkes, I., Oz-Levi, D., Alkelai, A., Olender, T., Zimmerman, S., Twik, M., Belinky, F., Fishilevich, S., and Nudel, R. (2016). VarElect: The phenotype-based variation prioritizer of the GeneCards Suite. BMC Genom., 17.","DOI":"10.1186\/s12864-016-2722-2"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"235","DOI":"10.1093\/nar\/28.1.235","article-title":"The Protein Data Bank","volume":"28","author":"Berman","year":"2000","journal-title":"Nucleic Acids Res."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"W296","DOI":"10.1093\/nar\/gky427","article-title":"SWISS-MODEL: Homology modelling of protein structures and complexes","volume":"46","author":"Waterhouse","year":"2018","journal-title":"Nucleic Acids Res."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"D439","DOI":"10.1093\/nar\/gkab1061","article-title":"AlphaFold Protein Structure Database: Massively expanding the structural coverage of protein-sequence space with high-accuracy models","volume":"50","author":"Varadi","year":"2022","journal-title":"Nucleic Acids Res."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"779","DOI":"10.1006\/jmbi.1993.1626","article-title":"Comparative protein modelling by satisfaction of spatial restraints","volume":"234","author":"Sali","year":"1993","journal-title":"J. Mol. Biol."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"1765","DOI":"10.1093\/bioinformatics\/btz828","article-title":"QMEANDisCo-distance constraints applied on model quality estimation","volume":"36","author":"Studer","year":"2020","journal-title":"Bioinformatics"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"W407","DOI":"10.1093\/nar\/gkm290","article-title":"ProSA-web: Interactive web service for the recognition of errors in three-dimensional structures of proteins","volume":"35","author":"Wiederstein","year":"2007","journal-title":"Nucleic Acids Res."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"W318","DOI":"10.1093\/nar\/gkae270","article-title":"The structure assessment web server: For proteins, complexes and more","volume":"52","author":"Waterhouse","year":"2024","journal-title":"Nucleic Acids Res."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"2577","DOI":"10.1002\/bip.360221211","article-title":"Dictionary of protein secondary structure: Pattern recognition of hydrogen-bonded and geometrical features","volume":"22","author":"Kabsch","year":"1983","journal-title":"Biopolymers"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"507","DOI":"10.1016\/0022-2836(91)90027-4","article-title":"Molecular recognition. Conformational analysis of limited proteolytic sites and serine proteinase protein inhibitors","volume":"220","author":"Hubbard","year":"1991","journal-title":"J. Mol. Biol."},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"International Multiple Sclerosis Genetics Consortium, Hafler, D.A., Compston, A., Sawcer, S., Lander, E.S., Daly, M.J., De Jager, P.L., de Bakker, P.I., Gabriel, S.B., and Mirel, D.B. (2007). Risk alleles for multiple sclerosis identified by a genomewide study. N. Engl. J. Med., 357, 851\u2013862.","DOI":"10.1056\/NEJMoa073493"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"328","DOI":"10.1016\/j.smim.2009.08.003","article-title":"Genetics and pathogenesis of multiple sclerosis","volume":"21","author":"Zuvich","year":"2009","journal-title":"Semin. Immunol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"59","DOI":"10.1186\/s13073-021-00878-y","article-title":"Emerging genetic complexity and rare genetic variants in neurodegenerative brain diseases","volume":"13","author":"Perrone","year":"2021","journal-title":"Genome Med."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"538","DOI":"10.1186\/s40001-024-02063-4","article-title":"Genes to therapy: A comprehensive literature review of whole-exome sequencing in neurology and neurosurgery","volume":"29","author":"Tan","year":"2024","journal-title":"Eur. J. Med. Res."},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"B\u00fcy\u00fckg\u00f6l, F., G\u00fcrdamar, B., Alu\u00e7lu, M.U., Beckmann, Y., Bilguvar, K., Boz, C., B\u00fclb\u00fcl, A., B\u00fcn\u00fcl, S.D., \u00c7etin, \u00d6., and Demir, C.F. (2025). Exome sequencing reveals low-frequency and rare variant contributions to multiple sclerosis susceptibility in Turkish families. Sci. Rep., 15.","DOI":"10.1038\/s41598-025-94691-x"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"150","DOI":"10.1016\/j.nrl.2022.10.001","article-title":"Exonic variants of the P2RX7 gene in familial multiple sclerosis","volume":"40","author":"Pytel","year":"2025","journal-title":"Neurologia"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"577760","DOI":"10.1016\/j.jneuroim.2021.577760","article-title":"Burden of rare coding variants in an Italian cohort of familial multiple sclerosis","volume":"362","author":"Mascia","year":"2022","journal-title":"J. Neuroimmunol."},{"key":"ref_31","doi-asserted-by":"crossref","unstructured":"Horjus, J., van Mourik-Banda, T., Heerings, M.A.P., Hakobjan, M., De Witte, W., Heersema, D.J., Jansen, A.J., Strijbis, E.M.M., de Jong, B.A., and Slettenaar, A.E.J. (2022). Whole Exome Sequencing in Multi-Incident Families Identifies Novel Candidate Genes for Multiple Sclerosis. Int. J. Mol. Sci., 23.","DOI":"10.1101\/2022.02.28.22271609"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"1778","DOI":"10.3390\/cimb43030125","article-title":"Homozygosity Haplotype and Whole-Exome Sequencing Analysis to Identify Potentially Functional Rare Variants Involved in Multiple Sclerosis among Sardinian Families","volume":"43","author":"Fazia","year":"2021","journal-title":"Curr. Issues Mol. Biol."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"2645","DOI":"10.1016\/j.ygeno.2021.06.008","article-title":"Exome sequencing reveals novel rare variants in Iranian familial multiple sclerosis: The importance of POLD2 in the disease pathogenesis","volume":"113","author":"Salehi","year":"2021","journal-title":"Genomics"},{"key":"ref_34","doi-asserted-by":"crossref","unstructured":"Zrzavy, T., Leutmezer, F., Kristoferitsch, W., Kornek, B., Schneider, C., Rommer, P., Berger, T., and Zimprich, A. (2020). Exome-Sequence Analyses of Four Multi-Incident Multiple Sclerosis Families. Genes, 11.","DOI":"10.3390\/genes11090988"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"1178","DOI":"10.1111\/cns.13456","article-title":"Variants of genes encoding TNF receptors and ligands and proteins regulating TNF activation in familial multiple sclerosis","volume":"26","author":"Pytel","year":"2020","journal-title":"CNS Neurosci. Ther."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"e01272","DOI":"10.1002\/brb3.1272","article-title":"Exonic variants of genes related to the vitamin D signaling pathway in the families of familial multiple sclerosis using whole-exome next generation sequencing","volume":"9","author":"Pytel","year":"2019","journal-title":"Brain Behav."},{"key":"ref_37","doi-asserted-by":"crossref","unstructured":"Vilari\u00f1o-G\u00fcell, C., Zimprich, A., Martinelli-Boneschi, F., Herculano, B., Wang, Z., Matesanz, F., Urcelay, E., Vandenbroeck, K., Leyva, L., and Gris, D. (2019). Exome sequencing in multiple sclerosis families identifies 12 candidate genes and nominates biological pathways for the genesis of disease. PLoS Genet., 15.","DOI":"10.1371\/journal.pgen.1008180"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"909","DOI":"10.1177\/1352458518777202","article-title":"Linkage analysis and whole exome sequencing identify a novel candidate gene in a Dutch multiple sclerosis family","volume":"25","author":"Mescheriakova","year":"2019","journal-title":"Mult. Scler."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"406","DOI":"10.1212\/WNL.0b013e3182616fc4","article-title":"Exome sequencing identifies a novel multiple sclerosis susceptibility variant in the TYK2 gene","volume":"79","author":"Dyment","year":"2012","journal-title":"Neurology"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"881","DOI":"10.1002\/ana.22678","article-title":"Rare variants in the CYP27B1 gene are associated with multiple sclerosis","volume":"70","author":"Ramagopalan","year":"2011","journal-title":"Ann. Neurol."},{"key":"ref_41","doi-asserted-by":"crossref","unstructured":"Kalafatakis, I., Papagianni, F., Theodorakis, K., and Karagogeos, D. (2023). Nogo-A and LINGO-1: Two Important Targets for Remyelination and Regeneration. Int. J. Mol. Sci., 24.","DOI":"10.3390\/ijms24054479"},{"key":"ref_42","doi-asserted-by":"crossref","unstructured":"Kulczy\u0144ska-Przybik, A., Mroczko, P., Dulewicz, M., and Mroczko, B. (2021). The Implication of Reticulons (RTNs) in Neurodegenerative Diseases: From Molecular Mechanisms to Potential Diagnostic and Therapeutic Approaches. Int. J. Mol. Sci., 22.","DOI":"10.3390\/ijms22094630"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"65","DOI":"10.1016\/j.neuroscience.2024.06.009","article-title":"JAK2\/STAT3 Pathway Inhibition by AG490 Ameliorates Experimental Autoimmune Encephalomyelitis via Regulation of Th17 Cells and Autophagy","volume":"552","author":"Xue","year":"2024","journal-title":"Neuroscience"},{"key":"ref_44","doi-asserted-by":"crossref","unstructured":"Tarafdar, A., and Pula, G. (2018). The Role of NADPH Oxidases and Oxidative Stress in Neurodegenerative Disorders. Int. J. Mol. Sci., 30.","DOI":"10.3390\/ijms19123824"}],"container-title":["Genes"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2073-4425\/16\/11\/1311\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,11,3]],"date-time":"2025-11-03T13:32:12Z","timestamp":1762176732000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2073-4425\/16\/11\/1311"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,11,1]]},"references-count":44,"journal-issue":{"issue":"11","published-online":{"date-parts":[[2025,11]]}},"alternative-id":["genes16111311"],"URL":"https:\/\/doi.org\/10.3390\/genes16111311","relation":{},"ISSN":["2073-4425"],"issn-type":[{"value":"2073-4425","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,11,1]]}}}