Plasma microRNAs as prognostic biomarkers for development of severe epilepsy after experimental traumatic brain injury—EpiBioS4Rx Project 1 study

Mette Heiskanen, Xavier Ekolle Ndode-Ekane, Idrish Ali, Cesar Santana-Gomez, Noora Puhakka, Shalini Das Gupta, Pedro Andrade, Riikka Immonen, Pablo Casillas-Espinosa, Eppu Manninen, Gregory Smith, Rhys D. Brady, Juliana Silva, Emma Braine, Matt Hudson, Glen R. Yamakawa, Nigel C. Jones, Sandy R. Shultz, Neil G. Harris, David K. WrightOlli Gröhn, Richard J. Staba, Terence J. O'Brien, Asla Pitkänen

Research output: Contribution to journalArticleResearchpeer-review

Abstract

Objective: To test a hypothesis that acutely regulated plasma microRNAs (miRNAs) can serve as prognostic biomarkers for the development of post-traumatic epilepsy (PTE). Methods: Adult male Sprague–Dawley rats (n = 245) were randomized to lateral fluid-percussion–induced traumatic brain injury (TBI) or sham operation at three study sites (Finland, Australia, United States). Video-electroencephalography (vEEG) was performed on the seventh post-injury month to detect spontaneous seizures. Tail vein plasma collected 48 h after TBI for miRNA analysis was available from 209 vEEG monitored animals (45 sham, 164 TBI [32 with epilepsy]). Based on small RNA sequencing and previous data, the seven most promising brain enriched miRNAs (miR-183-5p, miR-323-3p, miR-434-3p, miR-9a-3p, miR-124-3p, miR-132-3p, and miR-212-3p) were validated by droplet digital polymerase chain reaction (ddPCR). Results: All seven plasma miRNAs differentiated between TBI and sham-operated rats. None of the seven miRNAs differentiated TBI rats that did and did not develop epilepsy (p >.05), or rats with ≥3 vs <3 seizures in a month (p >.05). However, miR-212-3p differentiated rats that developed epilepsy with seizure clusters (i.e., ≥3 seizures within 24 h) from those without seizure clusters (.34 ±.14 vs.60 ±.34, adj. p <.05) with an area under the curve (AUC) of.81 (95% confidence interval [CI].65–.97, p <.01, 64% sensitivity, 95% specificity). Lack of elevation in miR-212-3p also differentiated rats that developed epilepsy with seizure clusters from all other TBI rats (n = 146,.34 ±.14 vs.55 ±.31, p <.01) with an AUC of.74 (95% CI.61–.87, p <.01, 82% sensitivity, 62% specificity). Glmnet analysis identified a combination of miR-212-3p and miR-132-3p as an optimal set to differentiate TBI rats with vs without seizure clusters (cross-validated AUC.75, 95% CI.47–.92, p <.05). Significance: miR-212-3p alone or in combination with miR-132-3p shows promise as a translational prognostic biomarker for the development of severe PTE with seizure clusters.

Original languageEnglish
Pages (from-to)870-885
Number of pages16
JournalEpilepsia
Volume205
DOIs
Publication statusPublished - Mar 2025

Keywords

  • harmonization
  • lateral fluid-percussion
  • post-traumatic epilepsy
  • preclinical
  • receiver-operating characteristic analysis
  • video-EEG monitoring

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