ARHGEF26 Maintains SOX2 Stability by Inhibiting Ubiquitination to Enhance Glioblastoma Stemness

Cancer stem cells represent a critical cell population that drives the malignant proliferation and invasiveness of glioblastoma (GBM), contributing to its poor prognosis. However, the mechanisms underlying the maintenance of stemness in GBM are poorly understood. This study aimed to investigate the role of Rho guanine nucleotide exchange factor 26 (ARHGEF26) in regulating GBM stemness and its underlying mechanism.

Human GBM specimens and non-tumor brain tissues were collected with ethical approval. GBM cell lines, and GBM stem cells (enriched by CD133+/CD15+ sorting) were cultured. We identified Rho guanine nucleotide exchange factor 26 (ARHGEF26) as a protein highly enriched in GBM stem cells and GBM tissues. Clinical correlation analysis was performed to assess the association between ARHGEF26 expression and patient survival. Gain-of-function and loss-of-function experiments were used to evaluate the effects of ARHGEF26 on GBM cell self-renewal, invasion, and tumorigenesis both in vitro and in vivo. Co-immunoprecipitation and ubiquitination assays were conducted to explore the molecular mechanism of ARHGEF26 in regulating GBM stemness. Bioinformatics analyses used data from TCGA, CGGA, Ivy Glioblastoma Atlas Project, and GlioVis. Experiments were repeated three times with GraphPad Prism-based statistical analyses.

ARHGEF26 was significantly enriched in GBM stem cells and GBM tissues, and its high expression correlated with shorter overall survival in GBM patients. ARHGEF26 overexpression enhanced the self-renewal, invasion, and tumorigenesis of GBM cells both in vitro and in vivo. Mechanistically, ARHGEF26 interacted with and stabilized the core stemness transcription factor SOX2 by reducing its K48-linked polyubiquitination and subsequent proteasomal degradation.

Our findings reveal a novel role and mechanism for ARHGEF26 in promoting GBM stemness and suggest its potential as a therapeutic target.