Authors :
Presenting Author: Zita Goliášová, Mgr. – Brno Epilepsy Centre, 1st Department of Neurology, St. Anne’s University Hospital and Faculty of Medicine, Masaryk University, Brno, Czech Republic
Jalleh Shakerzadeh, PhD – Brno Epilepsy Centre, 1st Department of Neurology, St. Anne’s University Hospital and Faculty of Medicine, Masaryk University, Brno, Czech Republic
Radim Jaroušek, Mgr. – Department of Experimental Biology, Faculty of Science, Masaryk University, Brno, 62500, Czech Republic
Milan Brázdil, MD, PhD – 1st Department of Neurology, Faculty of Medicine, Masaryk University and St. Anne´s University Hospital, Brno, Czech Republic – member of ERN EpiCARE
Hana Kubova, PhD – Institute of Physiology, Academy of Science, Czech Republic
Rationale:
Early-onset epilepsy presents a critical challenge to brain and long-term neurological outcomes, particularly during the vulnerable stages of early development. During this phase, the brain is highly plastic and susceptible to long-term alterations. However, the majority of the research focuses on the chronic phases of epilepsy, predominantly in mature animals. Instead, this study aims to fill the gap by elucidating the molecular mechanisms driving the acute phase of seizure onset in early development, potentially resulting in epileptogenesis-related remodelling. This objective was achieved using an integrative multi-omics analysis to comprehensively profile the molecular changes in transcriptomic and proteomic levels associated with acute seizures in a neonatal rat model. The presented results will elucidate the molecular mechanism during a critical window of brain development and highlight the new therapeutic molecules for treating temporal lobe epilepsy.
Methods:
Experiments were performed on twelve-day-old (P12) Wistar albino rats, including both sexes. Each group had 12 animals (6 control, 6 treatment). At P11, rats received an intraperitoneal injection of LiCl (127 mg/kg), followed by pilocarpine (35 mg/kg) 24 hours later to induce seizures. To reduce mortality, paraldehyde (0.07 ml/kg) was administered 1.5 hours post-seizure onset. Controls received saline instead of pilocarpine. Rats were sacrificed in three intervals – 24 hours, 7 days and 3 months after seizures. Hippocampi were dissected and frozen, and next-generation sequencing (mRNA and microRNA) and proteomics analyses (LC/MS) were performed. Validations were processed by the qRT-PCR method.
Results:
The analysis identified 11 overlapping dysregulated molecules involved in structural remodelling, neuroprotection, and maintenance of neuronal integrity, which were significantly dysregulated 24 hours post-seizure in neonatal rats. RNA sequencing revealed that specific microRNAs, for example, 155-5p or 132-5p, regulating these genes were also dysregulated, suggesting an alteration in controlling critical pathways during the acute phase of seizures. qRT-PCR confirmed these results in the biological validation cohort.
Conclusions:
The combined proteomic and transcriptomic findings highlight a coordinated alteration of inflammatory, stress, and remodelling pathways in early-onset epilepsy, driven by the dysregulation of essential genes and their regulatory microRNAs. These results provide potential biomarkers and therapeutic targets, offering new insights into the molecular mechanisms underlying acute seizures.
Funding:
Supported by project nr. LX22NPO5107 (MEYS): Financed by European Union – Next Generation EU.
This work has received funding from the Czech Science Foundation, project no. 23-05274S