A Single-Cell Transcriptomic Atlas Elucidates a Microglial Gene Signature Linking Ferroptosis to Mitochondrial Dysfunction in Epilepsy.

Background: Epilepsy is a chronic neurological disorder caused by abnormal synchronous discharges of neurons, with ferroptosis and mitochondrial dysfunction implicated in its progression. However, a lack of resolution at the level of specific cell types has obscured critical roles and molecular mechanisms within the epileptic microenvironment. Methods: This study integrated single-nucleus RNA sequencing (snRNA-seq) and bulk RNA-seq data from epilepsy and control samples. We performed dimensionality reduction and clustering to identify 13 cell subpopulations, and then assessed the expression of ferroptosis-related genes (FRGs) and mitochondrial-related genes (MRGs) within 7 major cell types. Three machine learning algorithms were further applied to identify key genes in microglia. Subsequent analyses included immune infiltration, pathway enrichment, and drug-target interaction. Molecular docking and molecular dynamics simulations were used to evaluate the potential binding affinity of the predicted drug. In vivo validation included histopathology, mitochondrial electron microscopy, JC-1 staining, and immunohistochemistry (IHC). Results: We identified six key genes ( Conclusion: This study highlights the interplay between ferroptosis and mitochondrial dysfunction in epilepsy, identifying six key microglial genes as high-priority candidates for future mechanistic and therapeutic investigation. Notably,