Journal Article Multicenter Study Ppublish
PubMed: 42789917 DOI: 10.1212/NXI.0000000000200659
Epigenetic Insights in Pediatric Multiple Sclerosis: DNA Methylation Highlights an Involvement of Epstein-Barr Virus Infection and Host Immune Networks.
Neurol Neuroimmunol Neuroinflamm 13(6):e200659
Authors: Andrea Corona, Martina Tosi, Alen Zollo, Nadia Barizzone, Nicola Pomella, Marta Simone, Alessandra Protti, Angela Lucia Berardinelli, Antonio Gallo, Carlotta Canavese, Domizia Vecchio, Eleonora Cocco, Lucia Moiola, Marta Zaffira Conti, Simona Malucchi, Amanda Papa, Alessandra Mingione, Corinne Monzani, Pietro Annovazzi, Luigi M E Grimaldi, Roberta Lanzillo, Sarah Rasia, Stefania Maria Bova, Paolo Ravanini, Fjorilda Caushi, Valentina Liliana Adriana Maria Torri Clerici, Stefano Sotgiu, Alberto Priori, Maria Trojano, Maria Pia Amato, Roberto Bergamaschi, Silvy Pilotto, Carlo Pozzilli, Salvatore Cottone, Giuseppe Santangelo, Giovanna De Luca, Maura Pugliatti, Angelo Ghezzi, Sandra D'Alfonso, Filippo Martinelli Boneschi
Affiliations
- Laboratory of Precision Medicine of Neurological Diseases, Department of Health Science, University of Milan, Italy.
- CRC "Aldo Ravelli" for Experimental Brain Therapeutics, Department of Health Science, University of Milan, Italy.
- Department of Health Sciences, University of Eastern Piedmont, Novara, Italy.
- Child Neuropsychiatric Unit, Department of Precision and Regenerative Medicine and Jonic Area, University of Bari Aldo Moro, Italy.
- Multiple Sclerosis Centre, Department of Neurosciences, ASST Niguarda Hospital, Milan, Italy.
- IRCCS Mondino Foundation, Pavia, Italy.
- Department of Advanced Medical and Surgical Sciences, University of Campania "Luigi Vanvitelli", Naples, Italy.
- Regina Margherita Children's Hospital, Città della Salute e della Scienza di Torino, Pediatric Neurology Unit, Turin, Italy.
- Department of Neurology, University Hospital "Maggiore della Carità", Novara, Italy.
- Multiple Sclerosis Center, ASL Cagliari, Department of Medical Sciences and Public Health, Binaghi Hospital, University of Cagliari, Italy.
- San Raffaele Hospital, Milan, Italy.
- ASST Papa Giovanni XXIII, Bergamo, Italy.
- Department of Neurology and Centri di Riferimento Regionale Sclerosi Multipla, University Hospital San Luigi Gonzaga, Orbassano, Italy.
- Child Neurology and Psychiatry Unit, University Hospital "Maggiore della Carità", Novara, Italy.
- Neuroimmunology Unit-MS Center, ASST Valle Olona, Gallarate Hospital, Gallarate (VA), Italy.
- Foundation Institute San Raffaele G. Giglio, Neurology Unit, Cefalù, Italy.
- UniCamillus - Saint Camillus International University of Health Sciences, Rome, Italy.
- Multiple Sclerosis Center, Department of Neuroscience, Reproductive Sciences and Odontostomatology, University of Naples "Federico II", Italy.
- Multiple Sclerosis Center, ASST Spedali Civili, PO di Montichiari (BS), Italy.
- V. Buzzi Children's Hospital, ASST Fatebenefratelli-Sacco, Pediatric Neurology, Milan, Italy.
- SCDO Microbiologia e Virologia, AOU Maggiore della Carità, Novara, Italy.
- Multiple Sclerosis Centre, C. Besta Neurological Institute, Milan, Italy.
- Child Neuropsychiatry Unit, Department of Medicine, Surgery and Pharmacy, University of Sassari, Italy.
- Clinical Neurology Unit, Azienda Socio-Sanitaria Territoriale Santi Paolo e Carlo, and Department of Health Sciences, University of Milan, Italy.
- Department of Translational Biomedicine and Neurosciences "DiBraiN", University of Bari Aldo Moro, Italy.
- Department of NEUROFARBA, University of Florence, Italy.
- Neurology Unit, San Martino Hospital, ASL 5 Oristano, Italy.
- Department of Human Neurosciences, Sapienza University of Rome, MS Center, Sant'Andrea Hospital, Italy.
- Department of Neurology, A.R.N.A.S. Civico, Palermo, Italy.
- Child Neurology and Psychiatry Department, Pediatric Hospital "G. Di Cristina"-ARNAS, Palermo, Italy.
- Neurology Unit, Univ. G. D'Annunzio, Policlinico SS Annunziata, Chieti, Italy.
- Department of Neuroscience and Rehabilitation, University of Ferrara, Italy; and.
- Translational Research Centre for Autoimmune and Allergic Diseases (CAAD), Novara, Italy.
Abstract
BACKGROUND AND OBJECTIVES
Pediatric-onset multiple sclerosis (PedMS) provides a unique opportunity to investigate MS pathogenesis because of the short interval between environmental exposures and disease onset. This study aimed to identify differentially methylated regions (DMRs) that mediate the interplay between environmental triggers and genomic factors in the early stages of the disease.
METHODS
In this multicenter study, we analyzed peripheral blood DNA from 175 Italian subjects (122 PedMS patients, 53 healthy controls) using the Infinium Methylation EPIC Array v2. All patients were enrolled within 4 years of onset. The bioinformatic pipeline included batch effect correction, normalization, and differential methylation analysis adjusted for sex and inferred cell-type proportions. DMRs were detected using DMRcate. Network analysis was performed by projecting DMR-overlapping genes onto the STRING interactome using the NDEx tool, followed by pathway enrichment analysis with gProfiler. To ensure robustness, we performed an internal 1/3-2/3 validation and sensitivity analyses on treatment-naïve and low-efficacy treatment (LETA) subgroups.
RESULTS
Our analysis revealed 55 DMRs, with 28 affecting promoter regions. Top signals involved genes such as CHI3L2, LPCAT1, CD19, PARP1, CPT1B-CHKB, and IGF2R, including several MS-associated loci. Signals mapping to CHI3L2/DENND2D and LPCAT1 were highly resilient, persisting across all validation and sensitivity subsets. Network analysis identified a subnetwork of 165 nodes where the Epstein-Barr virus (EBV) infection emerged as the top-ranked enriched pathway (p = 3.23 *10 -40). High-centrality regulatory hubs included PARP1 and IGF2R, the latter overlapping with established imprinting control regions.
DISCUSSION
Our findings identify a distinct epigenetic signature in early-stage PedMS, suggesting that DNA methylation changes are not stochastic but converge on high-centrality regulatory hubs. The stable association with EBV-related pathways and imprinting loci reinforces the hypothesis that epigenetic remodeling serves as a critical interface between early-life environmental exposures and genetic susceptibility in MS onset.
MeSH
Environmental Exposure, Sclerosis, Genes, Multiple Sclerosis, Promoter Regions, Genetic, Dna Methylation, Herpesvirus 4, Human, Viruses, Therapeutics, Epstein-Barr Virus Infections, Infections, Virus Diseases