Abstract

Background: Glioblastoma multiforme (GBM) is a highly aggressive primary brain tumor characterized by rapid proliferation, profound invasiveness, and resistance to conventional therapies. Deubiquitinating enzymes (DUBs), crucial regulators of protein homeostasis, have recently been implicated in GBM pathogenesis. However, the specific DUBs that play central roles in GBM pathogenesis and their exact molecular mechanisms remain to be further elucidated. Methods: We systematically analyzed GBM datasets and clinical samples to identify differentially expressed DUBs. Functional experiments, including genetic manipulation, immunoprecipitation coupled mass spectrometry (IP-MS), comprehensive metabolic assays, mitochondrial function assessments, and orthotopic mouse models, were conducted. Results: Here, we identified PSMD14 as a protein significantly upregulated in GBM, with a close correlation to poor prognosis of patients. Mechanistic exploration revealed that PSMD14 stabilized IMPDH2, the rate-limiting enzyme of purine nucleotide biosynthesis, by selectively removing K48-linked polyubiquitin chains. When PSMD14 is inhibited genetically or pharmacologically, IMPDH2 stability diminishes, causing impaired nucleotide metabolism, mitochondrial dysfunction, increased DNA damage signaling, and reduced tumor malignancy. Importantly, these metabolic issues can be reversed by exogenous guanosine, highlighting the key role PSMD14 in metabolic regulation. In translational medicine, the PSMD14 inhibitor, Thiolutin, curbed GBM progression in vitro and in vivo by disrupting the de novo purine biosynthesis and resulting in mitochondrial fragmentation. Moreover, Thiolutin synergized with TMZ to overcome resistance and boost efficacy. This study reveals a new GBM metabolic axis and presents a promising PSMD14-targeting therapy. Conclusions: PSMD14-IMPDH2 axis serves as a crucial