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TNFα protein, DNA methylation, mRNA and miRNA expression evaluation in multiple sclerosis.

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Summary

Tumor necrosis factor alpha (TNFα) is elevated in relapsing-remitting multiple sclerosis (RRMS) due to promoter hypomethylation and reduced miR-130a-3p levels. Saliva shows promise as a noninvasive biomarker for RRMS.

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DNA methylationTNFαepigenetic regulationmiR-130a-3pmultiple sclerosissaliva biomarkers

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Area of Science:

  • Immunology
  • Neuroscience
  • Epigenetics

Background:

  • Tumor necrosis factor alpha (TNFα) is a critical cytokine in multiple sclerosis (MS) pathogenesis, influencing inflammation and neurodegeneration.
  • TNFα expression is regulated by epigenetic (e.g., promoter methylation) and post-transcriptional (e.g., miRNA) mechanisms.
  • Relapsing-remitting MS (RRMS) involves complex molecular pathways requiring further investigation for diagnostic markers.

Purpose of the Study:

  • To investigate TNFα expression, promoter methylation, and miR-130a-3p regulation in RRMS patients.
  • To evaluate serum and saliva as potential diagnostic biofluids for RRMS.
  • To explore the interplay between epigenetic and post-transcriptional regulation of TNFα in RRMS.

Main Methods:

  • Quantification of TNFα protein and mRNA in serum, PBMCs, and saliva via ELISA and qPCR.
  • Analysis of TNFα promoter methylation using pyrosequencing.
  • Assessment of miR-130a-3p expression in serum and saliva via qPCR.
  • Bioinformatic analysis to predict miRNA-TNFα interactions.

Main Results:

  • RRMS patients exhibited significantly higher TNFα levels (mRNA and protein) compared to healthy controls (HC).
  • A significant hypomethylation of the TNFα promoter was observed in PBMCs of RRMS patients.
  • miR-130a-3p was markedly downregulated in both serum and saliva of RRMS patients, inversely correlating with TNFα expression.
  • Salivary TNFα levels paralleled serum alterations, indicating saliva's potential as a biomarker source.

Conclusions:

  • TNFα upregulation in RRMS is linked to promoter hypomethylation and decreased miR-130a-3p, suggesting coordinated epigenetic and post-transcriptional control.
  • The parallel findings in saliva and serum support the use of salivary TNFα and miR-130a-3p as minimally invasive biomarkers for MS monitoring and diagnosis.