Abstract
Glioblastoma (GBM) frequently activates hypoxia signaling even under normoxic conditions, yet the mechanism sustaining hypoxia-inducible factor-1α (HIF-1α) stability remains unclear. Here, we identify the E3 ubiquitin ligase TRIM25 as a key driver of this phenomenon. TRIM25, aberrantly upregulated in GBM, directly binds HIF-1α and catalyzes K11/K29-linked polyubiquitination at lysine 532 of hydroxylated HIF-1α, preventing its canonical proteasomal degradation. This non-canonical ubiquitin modification stabilizes HIF-1α despite normal oxygen availability and sustains a pseudohypoxic transcriptional program in GBM cells. Functional studies in GBM cell lines, patient-derived cultures, and tumor models demonstrate that TRIM25-mediated HIF-1α stabilization promotes tumor proliferation, invasion, and angiogenic potential. Importantly, small-molecule screening identified T7117 as an inhibitor that disrupts the TRIM25–HIF-1α interaction, suppresses tumor growth, and enhances temozolomide efficacy. Together, our findings uncover a previously unrecognized ubiquitin mechanism that stabilizes hydroxylated HIF-1α under normoxia, revealing the TRIM25–HIF-1α axis as a driver of GBM pseudohypoxia and a potential therapeutic target.
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Data availability
The RNA-seq dataset was deposited in the Sequence Read Archive (SRA) repository at NCBI under the accession number PRJNA1347646. The datasets used and analyzed during the current study are available from the corresponding author upon reasonable request.
Code availability
Public R packages used in this study include DESeq2 and edgeR. In-house Perl scripts for raw FASTQ sequencing data trimming are available from the corresponding author upon reasonable request, with no restrictions on access.
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The authors thank the MogoEdit group (https://www.mogoedit.com) for the assistance in language editing.
Funding
J.J. discloses support for the research of this work from the Natural Science Foundation of China (NSFC) (Grant numbers: 82472723 and 82172955). J. Shao discloses support for the research of this work from the Key Project of the Jiangsu Provincial Health Commission (Grant number: ZD2022038). H. H. discloses support for the research of this work from the 2024 Jiangsu Province Graduate Student Practice Program (Grant number: SJCX24_0728). Other authors declare no relevant funding.
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HH, KN, CL and MC contributed equally to this work. HH, KN, CL, and MC contributed to the design of the study, analyzed data, performed all experiments, prepared figures, and edited the manuscript. YL, JZ, YS and YC performed computational analyses. JS, JShao, Y L, JW and JJ conceptualized and coordinated the study, analyzed the data, and wrote the paper. All authors read and edited the manuscript.
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All methods and experimental protocols in this study were performed in accordance with the relevant guidelines and regulations (including the Declaration of Helsinki for human studies and the ARRIVE guidelines for animal research). For human-related research, approval was obtained from the Institutional Review Board and Ethics Committee of the Affiliated Wuxi People’s Hospital of Nanjing Medical University (Reference number: (2023)109). Written informed consent was obtained from all participants prior to their inclusion in the study. Furthermore, for any identifiable images originating from human research participants, the authors have obtained separate written informed consent for publication of these images. For animal-related research, all procedures involving live vertebrates were approved by the Ethics Committee of the Affiliated Wuxi People’s Hospital of Nanjing Medical University (Reference number: (2023)114) and were performed in strict compliance with the institutional animal care regulations and the ARRIVE guidelines.
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Huang, H., Ni, K., Li, C. et al. TRIM25 promotes glioblastoma progression by stabilizing HIF-1α expression in normoxia through K11/K29 polyubiquitination. Cell Death Dis (2026). https://doi.org/10.1038/s41419-026-08757-3
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DOI: https://doi.org/10.1038/s41419-026-08757-3
