Identification of Injury-Response State Determinants in Glioblastoma Stem Cells

• DOI: 10.1101/2025.04.07.647626
• Publication Date: 2025-04-13T06:15:11Z
• Authors: Fatemeh Molaei; Graham MacLeod; Shahan Haider; Aleczandria Tiffany; Frank M Oteng; Jacob M. Berman; Arianna Skirzynska; Molly S. Shoichet; Daniel Schramek; Peter B. Dirks; Stephane Angers

AbstractGlioblastoma (GBM) is a common and highly lethal type of primary brain tumor in adults. Therapeutic failure is partly attributed to a fraction of Glioblastoma Stem Cells (GSCs) that show high levels of heterogeneity and plasticity. GSCs exist in a transcriptional gradient between two states: Developmental (Dev) and Injury Response (IR) in which IR-GSCs exhibit more invasive behaviors. While previous studies have identified fitness genes in GSCs, the genes required to establish and maintain the Dev and IR states remain poorly defined. To identify the regulators of the IR GSC state, we performed a phenotypic genome-wide CRISPR-Cas9 knockout (KO) screen in patient-derived GSCs based on cell surface expression of the IR marker CD44. Notably, we found that perturbations of the histone acetyltransferaseEP300in IRGSCsled to decreased CD44 cell surface expression, significant downregulation of gene expression signatures associated with the IR state, and to decreased self-renewal and invasion. Furthermore, genetic targeting ofEp300in a mouse GBM model delayed tumor initiation and/or progression. Collectively, our results establish EP300 as a regulator of the IR state in GSCs and provide a mechanistic basis for its therapeutic targeting in GBM.SignificanceA genome-wide phenotypic CRISPR-Cas9 screen in a patient-derived Glioblastoma Stem Cell line identified the genes required to maintain the Injury-Response cellular state, with a focus on the histone acetyl transferase geneEP300. This study suggests how therapeutic targeting of cellular state could reduce the aggressiveness of GBM tumors.