DNA demethylation modulates GLUT1, neurovascular metabolic targets and seizures in rats with cortical dysplasia post seizure induction
Abstract number :
3.052
Submission category :
1. Basic Mechanisms / 1D. Mechanisms of Therapeutic Interventions
Year :
2025
Submission ID :
844
Source :
www.aesnet.org
Presentation date :
12/8/2025 12:00:00 AM
Published date :
Authors :
Presenting Author: Chaitali Ghosh, PhD – Cleveland Clinic
Kathyn Bisaha, MS – Cleveland Clinic
Tiffany Bartel, BS – Cleveland Clinic
Imad Najm, MD – Cleveland Clinic
Rationale: Focal cortical dysplasia is a malformation of cortical development, which is the most common cause of medically refractory epilepsy and often linked with metabolic neurovascular challenges. Recently, we reported that glucose transporter 1 (GLUT1) and cerebral glucose hypometabolism in human FCD is associated with hypermethylation of key glucose regulatory genes. This study investigates the cause and effect of DNA demethylation on GLUT1 and neurovascular changes as a potential therapeutic intervention strategy in CD rats’ post-seizure induction.
Methods: We utilized an irradiation-induced rat model of congenital CD (first hit). Then, induced a seizure with a sub-dose of pentylenetetrazole (second hit). Seizures were graded via Racine score, followed with and without DNA methylation inhibitor, 5Aza treatment (n=8-12/ rats per group). We evaluated the impact of DNA methylation inhibitor decitabine/5-Aza-2-deoxycytidine on the enzyme DNA methyltransferase 1 (DNMT1) activity. The GLUT1 expression were assessed by immunohistochemistry. Levels of GLUT1, vascular endothelial growth factor (VEGFα), lactate transporter monocarboxylate transporter (MCT2), and mammalian target of rapamycin (mTOR) signaling were studied via western blot. In addition, the vascular density in the cortical areas were co-labeled by endothelial cell marker, RECA1 and histological cresyl-violet staining for comparison.
Results: Our study revealed that DNA methylation inhibition in CD rats post-seizure induction, (1) decreased DNMT1 enzyme activity, and seizure scores (from Grade 4 to 2) with 5Aza treatment, 2) upregulated GLUT1 to be more comparable to normal levels in the cortex, 3) decreased VEGFα, MCT2 expression and mTOR signaling pathways in the 5Aza treatment group, and 4) returned vascular density to a more normalized pattern.
Conclusions: Together, these findings confirm that demethylation strategy modulates GLUT1, and key neurovascular metabolic targets, decreases seizure score and alters mTOR regulatory pathways in CD rats post seizure induction. Therefore, DNA demethylation therapeutic intervention maybe a promising approach in managing seizures and regulating the GLUT1-mTOR signaling pathway affected by cerebral glucose metabolism in epilepsy with cortical dysplasia.
Funding: This work is supported in part by the National Institute of Neurological Disorders and Stroke/National Institutes of Health grants R01NS095825 and R01NS137230 (to C.G.) and funding from Cleveland Clinic, Neurological Institute Epilepsy Center.
Basic Mechanisms