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GPX4-AUTAC induces ferroptosis in breast cancer by promoting the selective autophagic degradation of GPX4 mediated by TRAF6-p62

Cell Death & Differentiation (2025)Cite this article

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Abstract

Emerging evidence indicates that activation of ferroptosis by inhibition of glutathione peroxidase 4 (GPX4) may be exploited as a therapeutic strategy to suppress tumor growth and progression. However, application of GPX4 inhibitors in cancer treatment is hampered by their poor selectivity, which results in unfavorable toxicity. Herein, we identified GPX4 as a candidate for the autophagy pathway. We showed that GPX4 is ubiquitinated by TNF receptor-associated factor 6 (TRAF6), which promotes its recognition by p62 and leads to its selective autophagic degradation. Utilizing targeted protein degradation (TPD) approach, we developed a GPX4-targeted AUTAC and demonstrated that GPX4-AUTAC promoted the ubiquitination of GPX4, and enhanced the binding with GPX4 and p62, leading to the selective autophagy-dependent degradation of GPX4. Furthermore, GPX4-AUTAC treatment strongly induced ferroptosis and exhibited potent anti-cancer activity against breast cancer in vitro, in vivo, and patient-derived organoids (PDOs). Combination treatment of GPX4-AUTAC with sulfasalazine, a ferroptotic inducer, or chemotherapy drugs showed a synergistic anti-cancer effect against breast cancer. These results uncover a new targeted degradation strategy for GPX4 by inducing selective autophagy and provide a rationale for the use of GPX4-AUTAC as a novel therapeutic approach to treatment of breast cancer.

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Fig. 1: High expression of GPX4 is associated with clinical progression and poor prognosis in breast cancer.
Fig. 2: Knockdown of GPX4 expression enhances the anti-tumor activity of ferroptosis inducer.
Fig. 3: p62 mediates the selective autophagic degradation of GPX4 in breast cancer.
Fig. 4: E3 ligase TRAF6 interacts with and ubiquitinates GPX4 to promote its autophagic degradation.
Fig. 5: GPX4-AUTAC accelerates GPX4 degradation in an autophagy-dependent manner.
Fig. 6: GPX4-AUTAC induces ferroptosis in breast cancer cells.
Fig. 7: GPX4-AUTAC exerts anti-cancer activity in vitro, in vivo, and in patient-derived organoids.
Fig. 8: Combination treatment of GPX4-AUTAC and sulfasalazine or chemotherapy exerts a synergistic anti-cancer effect against breast cancer.

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All data needed to evaluate the conclusions in the paper are present in the paper and/or the Supplementary Materials. Further inquiries can be directed to the corresponding author.

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Funding

This study was funded by the National Natural Science Foundation of China (Project No. 82373064 and 81972480 for YC), Hunan Provincial Natural Science Foundation (Project No. 2022JJ80106 for YC), and the Project of Innovation-driven Plan in Central South University (Project No. 1053320221775 for XW).

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Author notes

  1. These authors contributed equally: Rong Gong, Xiaoya Wan.

Authors and Affiliations

  1. Department of Pharmacy, The Second Xiangya Hospital, Central South University, Changsha, China

    Rong Gong, Xiaoya Wan, Yizhi Li, Ting Jiang, Changxin Zhong, Linhao He & Yan Cheng

  2. Hunan Provincial Engineering Research Centre of Translational Medicine and Innovative Drug, Changsha, China

    Rong Gong, Xiaoya Wan, Yizhi Li, Ting Jiang, Changxin Zhong, Linhao He & Yan Cheng

  3. Department of Pharmacy, Xiangya Hospital, Central South University, Changsha, China

    Shilong Jiang

  4. Department of Pathology, Xiangya Hospital, Central South University, Changsha, China

    Yidi Guan

  5. Department of General Surgery, The Second Xiangya Hospital, Central South University, Changsha, China

    Zonglin Chen

  6. Department of Laboratory Medicine, The Second Xiangya Hospital, Central South University, Changsha, China

    Zhongyuan Xiang

  7. Department of Medicinal Chemistry, School of Pharmacy, Chongqing Medical University, Chongqing, China

    Junya Yang & Biao Xu

  8. Department of Cancer Biology and Toxicology, Department of Pharmacology, College of Medicine and Markey Cancer Center, University of Kentucky, Lexington, KY, USA

    Jinming Yang

  9. FuRong Laboratory, Changsha, China

    Yan Cheng

  10. NHC Key Laboratory of Cancer Proteomics & State Local Joint Engineering Laboratory for Anticancer Drugs, Xiangya Hospital, Central South University, Changsha, China

    Yan Cheng

  11. Clinical Research Center for Breast Disease in Hunan Province, Changsha, China

    Yan Cheng

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Contributions

RG and XW contributed equally to this work. YC conceived and directed the project. RG and XW contributed to experimental design and program strategy. BX and JY contributed to design and chemical synthesis of compounds. RG and XW contributed to the in vitro biology experiments and data analysis. RG, XW, SJ, YG, YL, TJ, CZ, and LH contributed to the in vivo biology experiments and data analysis. ZY and ZC provided technological support in the experiments. RG and YC wrote the manuscript. JM and YC reviewed and edited the paper. All authors approved the final manuscript.

Corresponding author

Correspondence to Yan Cheng.

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The authors declare that they have no competing interests.

Ethics approval

All the animal experiments were approved by the Ethics Committee of the Department of Laboratory Animals of Central South University (code/CSU-2022-0001-0029), and the animal protocol was in accordance with the institutional guidelines of the Animal Care and Use Committee of Central South University. The maximal tumor burden allowed by ethics did not exceed 2000 mm3. The ethical endpoint was determined once the maximal tumor burden was reached. All the experiments performed with human specimens were reviewed and approved by the Research Ethics Board at the Second Xiangya Hospital for Cancer Research (code/2023-0233). All patients whose tumor samples were used in the study gave informed consent.

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Gong, R., Wan, X., Jiang, S. et al. GPX4-AUTAC induces ferroptosis in breast cancer by promoting the selective autophagic degradation of GPX4 mediated by TRAF6-p62. Cell Death Differ (2025). https://doi.org/10.1038/s41418-025-01528-1

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