Molecular and spatial heterogeneity of microglia in Rasmussen encephalitis

Rasmussen encephalitis (RE) is a rare childhood neurological disease characterized by progressive unilateral loss of function, hemispheric atrophy and drug-resistant epilepsy. Affected brain tissue shows signs of infiltrating cytotoxic T-cells, microglial activation, and neuronal death, implicating an inflammatory disease process. Recent studies have identified molecular correlates of inflammation in RE, but cell-type-specific mechanisms remain unclear. We used single-nucleus RNA-sequencing (snRNA-seq) to assess gene expression across multiple cell types in brain tissue resected from two children with RE. We found transcriptionally distinct microglial populations enriched in RE compared to two age-matched individuals with unaffected brain tissue and two individuals with Type I focal cortical dysplasia (FCD). Specifically, microglia in RE tissues demonstrated increased expression of genes associated with cytokine signaling, interferon-mediated pathways, and T-cell activation. We extended these findings using spatial proteomic analysis of tissue from four surgical resections to examine expression profiles of microglia within their pathological context. Microglia that were spatially aggregated into nodules had increased expression of dynamic immune regulatory markers (PD-L1, CD14, CD11c), T-cell activation markers (CD40, CD80) and were physically located near distinct CD4+ and CD8+ lymphocyte populations. These findings help elucidate the complex immune microenvironment of RE.

Automated summary

Rasmussen encephalitis (RE) is characterized by progressive loss of function and drug-resistant epilepsy, with affected brain tissue showing signs of inflammation. A recent study used single-nucleus RNA sequencing to examine gene expression in brain tissue from RE patients, revealing transcriptionally distinct microglial populations enriched in RE. These microglia showed increased expression of genes associated with cytokine signaling, interferon-mediated pathways, and T-cell activation. Furthermore, spatial proteomic analysis confirmed the presence of microglia with immune regulatory markers and their proximity to specific lymphocyte populations. This study provides insights into the complex immune microenvironment of RE.