Estrogen-related receptor beta activation and isoform shifting by cdc2-like kinase inhibition restricts migration and intracranial tumor growth in glioblastoma

Glioblastoma (GBM; grade 4 glioma) is a highly aggressive and incurable tumor. GBM has recently been characterized as highly dependent on alternative splicing, a critical driver of tumor heterogeneity and plasticity. Estrogen-related receptor beta (ERRβ, ESRRB, NR3B2 ) is an orphan nuclear receptor expressed in the brain, where alternative splicing of the 3’ end of the pre-mRNA leads to the production of three validated ERRβ protein products – ERRβ short form (ERRβsf), ERRβ2, and ERRβ exon 10-deleted (ERRβ-Δ10). Our prior studies have shown the ERRβ2 isoform to play a role in G2/M cell cycle arrest and induction of apoptosis, in contrast to the function of the shorter ERRβsf isoform in senescence and G1 cell cycle arrest. In this study, we sought to better define the role of the pro-apoptotic ERRβ2 isoform in GBM. We show that the ERRβ2 isoform is located in the nucleus, but also the cytoplasm. ERRβ2 suppresses GBM cell migration, interacts with the actin nucleation-promoting factor cortactin, and an ERRβ agonist is able to remodel the actin cytoskeleton and similarly suppress GBM cell migration. We further show that inhibition of the splicing regulatory cdc2-like kinases (CLKs) in combination with an ERRβ agonist shifts isoform expression in favor of ERRβ2 and potentiates inhibition of growth and migration in GBM cells and intracranial tumors.