ORIGINAL RESEARCH article
Front. Mol. Biosci.
Sec. Structural Biology
This article is part of the Research TopicTranslational Advances in Molecular Biomarkers for Disease Diagnosis and Clinical Monitoring: From Non-coding RNAs to Extracellular Vesicle ProteomicsView all articles
TNFα protein, DNA methylation, mRNA and miRNA expression evaluation in multiple sclerosis
Provisionally accepted
Erica Costantini1*
Nigel H Greig2
Lisa Aielli1
Paula Silva3Martina Di Carlo1Fani Konstantinidou1
Marcella Reale1- 1University of Studies G. d'Annunzio Chieti and Pescara, Chieti, Italy
- 2National Institute on Aging Intramural Research Program, Baltimore, United States
- 3Universidade do Porto Instituto de Ciencias Biomedicas Abel Salazar, Porto, Portugal
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Background: Tumor necrosis factor alpha (TNFα) is a key cytokine involved in the inflammatory and neurodegenerative processes underlying multiple sclerosis (MS). Its expression is finely regulated by epigenetic and post-transcriptional mechanisms, including promoter methylation and miRNA activity. The objective of this study is to investigate TNFα expression, promoter methylation, and its regulation by miR-130a-3p in patients with relapsing–remitting (RR)MS, evaluating both serum and saliva as potential diagnostic biofluids. Methods: RRMS patients in clinical remission and sex and age-matched healthy controls (HC) were enrolled. TNFα levels were quantified in serum, peripheral blood mononuclear cell (PBMC) supernatants, and saliva using ELISA. TNFα mRNA expression and promoter methylation were analyzed by qPCR and pyrosequencing, respectively. Bioinformatic tools (TargetScan, miRTargetLink 2.0, miEAA 2) were used to explore miRNA–TNFα interactions, and miR-130a-3p expression was evaluated in serum and saliva by qPCR. Results: RRMS patients showed significantly higher TNFα mRNA and protein levels compared to HC , paralleled by significant hypomethylation of the TNFα promoter in PBMCs. miR-130a-3p was markedly downregulated in both serum and saliva, exhibiting an inverse trend with TNFα expression. Salivary TNFα levels mirrored serum alterations, supporting the feasibility of saliva as a noninvasive biomarker source. Conclusions: The data indicate that TNFα upregulation in RRMS is associated with promoter hypomethylation and reduced miR-130a-3p expression, suggesting a coordinated epigenetic and post-transcriptional control of this cytokine. The parallel trends observed in saliva and serum highlight the potential use of salivary TNFα and miR-130a-3p as minimally invasive biomarkers for MS monitoring and early diagnosis.
Keywords: Multiple Sclerosis, TNFα, miR-130a-3p, DNA Methylation, saliva biomarkers, Epigenetic regulation
Received: 22 Oct 2025; Accepted: 25 Nov 2025.
Copyright: © 2025 Costantini, Greig, Aielli, Silva, Di Carlo, Konstantinidou and Reale. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
* Correspondence: Erica Costantini
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