305: Circulating microRNA Signatures for De Novo Pain Following SARS-CoV-2 Infection: A Multicenter Study

Topic: Clinical Science
Subtopic: Non-specific to any diagnosis, Omics (Proteomics/Metabolomics/Transcriptomics/Genomics), Genetics and Epigenetics

Background and Aims: Post-COVID pain is a global healthcare emergency affecting up to 60% of survivors, severely impacting quality of life. Distinguishing de novo post-COVID pain from pre-existing pain conditions is essential for clinical management. Recent research highlights that miRNA regulate host-virus interactions and can predict infection severity by modulating the overexpressed inflammatory response. As stable, circulating epigenetic regulators, miRNAs also influence inflammatory and nociceptive pathways. This study aims to evaluate plasma miRNA profiles in subjects reporting de novo post-COVID pain after SARS-CoV-2 infection, and the biological pathways they are regulating.

Methods: 287 COVID-19 survivors (197female,90male;age 46.5±15.1years) were recruited from two centers in Italy and Brazil. Post-infection assessments included questionnaires capturing pain before infection and current pain, together with a post-infection blood sample. Next-generation sequencing was used to identify differentially expressed miRNAs, validated by qRT-PCR (22 miRNAs). Based on questionnaire responses, participants were classified into pain phenotype groups: No pain (nopain before/no current pain), Persistent pain (pain before/current pain), and De novo pain (nopain before/current pain). T-test/Wilcoxon were used for miRNA expression evaluation between groups, Benjamini–Hochberg corrected(FDR<0.05). Pathway analysis was conducted using mirAnno.

Results: When comparing pain phenotypes, hsa-miR-30b-5p was found to be downregulated in De novo pain group (n=52) compared with No pain group (n=100) (Fold change (FC)=0.88, p_adj=0.015). In the comparison between Persistent pain group (n=56) vs No pain group (n=100), five miRNAs were significantly up/down-regulated: hsa-miR-146a-5p (FC=1.2895, p_adj =0.0014), hsa-miR-30b-5p (FC=0.8381, p_adj =0.0014), hsa-miR-320a-3p (FC=1.4999, p_adj =0.0016), hsa-miR-423-5p (FC=1.5, p_adj =0.0077), and hsa-miR-223-3p (FC=0.87, p_adj =0.0296). Pathway enrichment analysis identified 174 significantly enriched pathways targeted by the significant miRNAs, predominantly related to neuroinflammatory signaling and immune response regulation.

Conclusions: This multicenter study identifies hsa-miR-30b-5p as candidate biomarker across post-COVID pain trajectories, showing downregulation in both De novo and Persistent pain phenotypes compared with individuals reporting no pain. Additional dysregulation of hsa-miR-146a-5p, hsa-miR-320a-3p, hsa-miR-423-5p, and hsa-miR-223-3p in persistent pain suggests a broader molecular signature linked to pain maintenance. While these findings provide molecular evidence for epigenetic dysregulation in post-COVID pain, it remains to be determined whether they are virus-specific or reflect general molecular mechanisms underlying pain that develops after infection, stress, or acute disease.

References: Fernández-de-las-Peñas, C., Nijs, J., Giordano, R., & Arendt-Nielsen, L. (2023). Precision management of post-COVID pain: An evidence and clinical-based approach. European Journal of Pain, 27(9), 1107–1125.
Mustafa, F., Ahmad, W., Gull, B., Baby, J., Panicker, N. G., Khader, T. A., ... & Rizvi, T. A. (2025). miRNA biomarkers for prognosis and therapy monitoring in a multi-ethnic cohort with SARS-CoV-2 infection. Scientific Reports, 15.
Nicoletti, A. S., Visacri, M. B., Correa da Ronda, C. R. S., Siguemoto, J. T., Neri, C. M., de Souza, R. N., ... & Pincinato, E. C. (2024). Increased expression of miR-320b in blood plasma of patients in response to SARS-CoV-2 infection. Scientific Reports, 14.
Paval, N. E., Căliman-Sturdza, O. A., Lobiuc, A., Dimian, M., Sirbu, I. O., & Covasa, M. (2025). MicroRNAs in long COVID: roles, diagnostic biomarker potential and detection. Human Genomics, 19, 90.
Pawar, P., Akolkar, K., & Saxena, V. (2024). An integrated bioinformatics approach reveals the potential role of microRNA-30b-5p and let-7a-5p during SARS CoV-2 spike-1 mediated neuroinflammation. International Journal of Biological Macromolecules, 277, 134329.
Pius-Sadowska, E., Kulig, P., Niedźwiedź, A., Baumert, B., Rogińska, D., Łuczkowska, K., ... & Machaliński, B. (2025). The micro-RNA expression profile predicts the severity of SARS-CoV-2 infection. Scientific Reports.

Presenting Author: Rocco Giordano
Submitting Author: Rocco Giordano

Senior Author: Lars Arendt-Nielsen

IASP 2026 Global Year theme of Neuropathic Pain: No

• Gabriel Taricani Kubota: MD - Pain Center, Department of Neurology, University of Sao Paulo, Sao Paulo, Brazil.
• Caterina Leone: MD, Ph.D. - Department of Human Neuroscience, Sapienza University, Rome, Italy
• Daniel Ciampi de Andrade: MD, Dr med, Ph.D - Center for Neuroplasticity and Pain (CNAP), Department of Health and Technology, The Faculty of Medicine, Aalborg University, Denmark
• Andrea Truini: MD, Ph.D. - Department of Human Neuroscience, Sapienza University, Rome, Italy
• César Fernández De Las Peñas: PT, DO, MSc, DrMed, Ph.D. - Department of Physical Therapy, Occupational Therapy, Rehabilitation and Physical Medicine, Universidad Rey Juan Carlos, 28922 Alcorcón, Spain | Center for Neuroplasticity and Pain (CNAP), Department of Health and Technology, The Faculty of Medicine, Aalborg University, Denmark
• Lars Arendt-Nielsen: Dr med, Ph.D. - Center for Neuroplasticity and Pain (CNAP), Department of Health and Technology, The Faculty of Medicine, Aalborg University, Denmark