REST transcription factor holds the balance between the invasion and cell differentiation in IDH-mutant and IDH-wild type gliomas

Glioblastoma is the deadliest brain tumor with median survival of 15 months due to diffusive growth and lack of efficient therapy. Recurrent mutations in genes coding for isocitrate dehydrogenases (IDH) 1 or 2 cause to a change in function of the enzymes, resulting in production of 2-hydroxyglutarate, which leads to DNA hypermethylation phenotype, affecting transcription activators/repressors binding to DNA and gene expression. REST is a canonical repressor of neuronal genes in non-neuronal cells, whose levels differ between IDH wt and mut gliomas and whose expression levels correlate with the patients’ survival probability. IDH1 mutant-U-87 Isogenic Cell Line and malignant glioma U-87 MG were used in the experiments. For each cell line, REST-chromatin immunoprecipitation was performed twice, followed by sequencing and position weight matrix (PWM) analysis. Total RNA was extracted from U-87 MG and IDH1 mutant-U-87 isogenic cell lines that had not been treated, had control siRNA transfected, and had REST-targeting siRNA transfected. Following the construction and sequencing of mRNA sequencing libraries, differential analysis between REST knockdown and control samples was performed.