USP25 aggravates liver cancer development and impairs chemosensitivity by limiting LATS1 activation

Abstract Hepatocellular carcinoma (HCC) is a primary liver malignancy with high rates of morbidity and mortality, yet effective treatment options remain limited. Accumulating evidence indicates that large tumour suppressor kinase 1 (LATS1), a core kinase in the Hippo pathway, mediates the phosphorylation of downstream YAP/TAZ, thereby suppressing HCC progression. Although LATS1 protein stability has been reported to be modulated by ubiquitination, the specific mechanism controlling LATS1 kinase activity remains unknown. Here, we identify USP25 as a deubiquitinase that regulates LATS1 activity, independent of LATS1 protein stability. We demonstrated that USP25 is overexpressed in HCC and that USP25 depletion significantly suppresses HCC cell and tumour growth. Mechanistically, USP25 catalyses the removal of K63-linked ubiquitin at K688 of LATS1, which disrupts LATS1-MOB1 complex formation and promotes YAP-mediated transcriptional activation. Furthermore, we successfully developed a cell-penetrating peptide that disrupts the USP25-LATS1 interaction, which increases p-YAP expression and synergizes with chemotherapy in xenograft and patient-derived organoid and xenograft models. Collectively, our findings reveal a mechanism through which USP25 regulates LATS1 activity and provide a potential therapeutic strategy for HCC treatment.