KDM6A Loss Confers an Invasive Phenotype in Osteosarcoma by Activating Cytoplasmic YAP-Dependent β-catenin Stabilisation

Osteosarcoma (OS) is the most common primary malignant bone tumour and is characterised by aggressive growth, early metastasis, and a very stagnant clinical outcome. Although epigenetic dysregulation has been implicated in OS progression, the mechanisms linking epigenetic alterations to metastatic signalling remain unclear. Here, we identified the lysine-protein demethylase 6A (KDM6A/UTX) as a critical suppressor of OS metastasis and uncovered a novel regulatory axis involving the Hippo/YAP and Wnt/{beta}-catenin signalling. Initial bioinformatics analyses revealed frequent KDM6A alterations and significantly reduced expression in OS patient datasets, which correlated with metastatic disease and poor prognosis. Functional inhibition of KDM6A by pharmacological inhibitors and siRNA induced a hyper-invasive phenotype, marked by elevated mesenchymal markers, enhanced cytoskeletal remodelling, increased transendothelial adhesion and decreased chemotherapeutic drug sensitivity. Importantly, restoration of KDM6A expression effectively counteracted these effects. Mechanistically, KDM6A loss activated Wnt/{beta}-catenin signalling, resulting in nuclear translocation of {beta}-catenin and transcriptional activation of genes associated with stemness and invasion. Therefore, inhibition of {beta}-catenin reversed the invasive phenotype. Further analysis revealed that KDM6A regulated Hippo signalling through epigenetic control of the negative regulator of Yes-Associated Protein (YAP)-LATS1. KDM6A inhibition led to enrichment of H3K27me3, a repressive mark, at the LATS1 promoter. Accumulated YAP was predominantly localised in the cytoplasm, where it interacted with GSK3{beta} and contributed to the stabilisation of {beta}-catenin by preventing its proteasomal degradation. Collectively, our findings identify a novel KDM6A-LATS1-YAP-{beta}-catenin signalling axis that drives metastatic progression in OS.