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APE1 inhibition enhances ferroptotic cell death and contributes to hepatocellular carcinoma therapy

Cell Death & Differentiation volume 31, pages 431–446 (2024)Cite this article

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Abstract

Ferroptosis, a regulated form of cell death triggered by iron-dependent lipid peroxidation, has emerged as a promising therapeutic strategy for cancer treatment, particularly in hepatocellular carcinoma (HCC). However, the mechanisms underlying the regulation of ferroptosis in HCC remain to be unclear. In this study, we have identified a novel regulatory pathway of ferroptosis involving the inhibition of Apurinic/apyrimidinic endonuclease 1 (APE1), a key enzyme with dual functions in DNA repair and redox regulation. Our findings demonstrate that inhibition of APE1 leads to the accumulation of lipid peroxidation and enhances ferroptosis in HCC. At the molecular level, the inhibition of APE1 enhances ferroptosis which relies on the redox activity of APE1 through the regulation of the NRF2/SLC7A11/GPX4 axis. We have identified that both genetic and chemical inhibition of APE1 increases AKT oxidation, resulting in an impairment of AKT phosphorylation and activation, which leads to the dephosphorylation and activation of GSK3β, facilitating the subsequent ubiquitin-proteasome-dependent degradation of NRF2. Consequently, the downregulation of NRF2 suppresses SLC7A11 and GPX4 expression, triggering ferroptosis in HCC cells and providing a potential therapeutic approach for ferroptosis-based therapy in HCC. Overall, our study uncovers a novel role and mechanism of APE1 in the regulation of ferroptosis and highlights the potential of targeting APE1 as a promising therapeutic strategy for HCC and other cancers.

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Fig. 1: Inhibition of APE1 promotes ferroptosis in HCC cells.
Fig. 2: APE1 regulates ferroptosis through NRF2/SLC7A11/GPX4 axis.
Fig. 3: APE1 regulates the stability of NRF2 through GSK3β-involved ubiquitination/proteasome pathway.
Fig. 4: APE1-modulated ferroptosis is NRF2-depentent.
Fig. 5: The DNA damage repair function of APE1 is not involved in ferroptosis.
Fig. 6: APE1 relies on its redox function to participate in ferroptosis.
Fig. 7: APE1 inhibition enhances the sensitivity of HCC to ferroptosis in vivo.
Fig. 8: APE1-mediated expression of SLC7A11 and GPX4 via NRF2 is clinically associated with the progression and prognosis of HCC.

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All data needed to evaluate the conclusions in the paper are present in the paper. Additional data related to this paper may be requested from the corresponding author.

References

  1. Faivre S, Rimassa L, Finn RS. Molecular therapies for HCC: Looking outside the box. J Hepatol. 2020;72:342–52.

    Article  CAS  PubMed  Google Scholar 

  2. Dixon SJ, Lemberg KM, Lamprecht MR, Skouta R, Zaitsev EM, Gleason CE, et al. Ferroptosis: an iron-dependent form of nonapoptotic cell death. Cell. 2012;149:1060–72.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. Jiang L, Kon N, Li T, Wang SJ, Su T, Hibshoosh H, et al. Ferroptosis as a p53-mediated activity during tumour suppression. Nature. 2015;520:57–62.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. Wang W, Green M, Choi JE, Gijon M, Kennedy PD, Johnson JK, et al. CD8(+) T cells regulate tumour ferroptosis during cancer immunotherapy. Nature. 2019;569:270–4.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. Lang X, Green MD, Wang W, Yu J, Choi JE, Jiang L, et al. Radiotherapy and Immunotherapy Promote Tumoral Lipid Oxidation and Ferroptosis via Synergistic Repression of SLC7A11. Cancer Discov. 2019;9:1673–85.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Kagan VE, Mao G, Qu F, Angeli JP, Doll S, Croix CS, et al. Oxidized arachidonic and adrenic PEs navigate cells to ferroptosis. Nat Chem Biol. 2017;13:81–90.

    Article  CAS  PubMed  Google Scholar 

  7. Koppula P, Zhuang L, Gan B. Cystine transporter SLC7A11/xCT in cancer: ferroptosis, nutrient dependency, and cancer therapy. Protein Cell. 2021;12:599–620.

    Article  CAS  PubMed  Google Scholar 

  8. Yang WS, SriRamaratnam R, Welsch ME, Shimada K, Skouta R, Viswanathan VS, et al. Regulation of ferroptotic cancer cell death by GPX4. Cell. 2014;156:317–31.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Chen J, Li X, Ge C, Min J, Wang F. The multifaceted role of ferroptosis in liver disease. Cell Death Differ. 2022;29:467–80.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Liao H, Shi J, Wen K, Lin J, Liu Q, Shi B, et al. Molecular Targets of Ferroptosis in Hepatocellular Carcinoma. J Hepatocell Carcinoma. 2021;8:985–96.

    Article  PubMed  PubMed Central  Google Scholar 

  11. Chen Q, Zheng W, Guan J, Liu H, Dan Y, Zhu L, et al. SOCS2-enhanced ubiquitination of SLC7A11 promotes ferroptosis and radiosensitization in hepatocellular carcinoma. Cell Death Differ. 2023;30:137–51.

    Article  CAS  PubMed  Google Scholar 

  12. Wang Q, Bin C, Xue Q, Gao Q, Huang A, Wang K, et al. GSTZ1 sensitizes hepatocellular carcinoma cells to sorafenib-induced ferroptosis via inhibition of NRF2/GPX4 axis. Cell Death Dis. 2021;12:426.

    Article  PubMed  PubMed Central  Google Scholar 

  13. Bhakat KK, Mantha AK, Mitra S. Transcriptional regulatory functions of mammalian AP-endonuclease (APE1/Ref-1), an essential multifunctional protein. Antioxid Redox Signal. 2009;11:621–38.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Demple B, Herman T, Chen DS. Cloning and expression of APE, the cDNA encoding the major human apurinic endonuclease: definition of a family of DNA repair enzymes. Proc Natl Acad Sci USA. 1991;88:11450–4.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Li M, Wilson DM 3rd. Human apurinic/apyrimidinic endonuclease 1. Antioxid Redox Signal. 2014;20:678–707.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. Kelley MR, Georgiadis MM, Fishel ML. APE1/Ref-1 role in redox signaling: translational applications of targeting the redox function of the DNA repair/redox protein APE1/Ref-1. Curr Mol Pharm. 2012;5:36–53.

    Article  CAS  Google Scholar 

  17. Nishi T, Shimizu N, Hiramoto M, Sato I, Yamaguchi Y, Hasegawa M, et al. Spatial redox regulation of a critical cysteine residue of NF-kappa B in vivo. J Biol Chem. 2002;277:44548–56.

    Article  CAS  PubMed  Google Scholar 

  18. Logsdon DP, Grimard M, Luo M, Shahda S, Jiang Y, Tong Y, et al. Regulation of HIF1alpha under Hypoxia by APE1/Ref-1 Impacts CA9 Expression: Dual Targeting in Patient-Derived 3D Pancreatic Cancer Models. Mol Cancer Ther. 2016;15:2722–32.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. Ando K, Hirao S, Kabe Y, Ogura Y, Sato I, Yamaguchi Y, et al. A new APE1/Ref-1-dependent pathway leading to reduction of NF-kappaB and AP-1, and activation of their DNA-binding activity. Nucleic Acids Res. 2008;36:4327–36.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  20. Seo YR, Kelley MR, Smith ML. Selenomethionine regulation of p53 by a ref1-dependent redox mechanism. Proc Natl Acad Sci USA. 2002;99:14548–53.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Caston RA, Gampala S, Armstrong L, Messmann RA, Fishel ML, Kelley MR. The multifunctional APE1 DNA repair-redox signaling protein as a drug target in human disease. Drug Discov Today. 2021;26:218–28.

    Article  CAS  PubMed  Google Scholar 

  22. Lu X, Zhao H, Yuan H, Chu Y, Zhu X. High nuclear expression of APE1 correlates with unfavorable prognosis and promotes tumor growth in hepatocellular carcinoma. J Mol Histol. 2021;52:219–31.

    Article  CAS  PubMed  Google Scholar 

  23. Li E, Xia M, Du Y, Long K, Ji F, Pan F, et al. METTL3 promotes homologous recombination repair and modulates chemotherapeutic response in breast cancer by regulating the EGF/RAD51 axis. Elife. 2022;11:e75231.

  24. Yang F, Hu Z, Guo Z. Small-Molecule Inhibitors Targeting FEN1 for Cancer Therapy. Biomolecules. 2022;12:1007.

  25. Ji F, Zhang F, Zhang M, Long K, Xia M, Lu F, et al. Targeting the DNA damage response enhances CD70 CAR-T cell therapy for renal carcinoma by activating the cGAS-STING pathway. J Hematol Oncol. 2021;14:152.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  26. Long K, Gu L, Li L, Zhang Z, Li E, Zhang Y, et al. Small-molecule inhibition of APE1 induces apoptosis, pyroptosis, and necroptosis in non-small cell lung cancer. Cell Death Dis. 2021;12:503.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  27. Zhao H, Ding Y, Zhang L. SIRT1/APE1 promotes the viability of gastric cancer cells by inhibiting p53 to suppress ferroptosis. Open Med (Wars). 2023;18:20220620.

    Article  CAS  PubMed  Google Scholar 

  28. Yue R, Zhou M, Li X, Xu L, Lu C, Dong Z, et al. GSH/APE1 Cascade-Activated Nanoplatform for Imaging Therapy Resistance Dynamics and Enzyme-Mediated Adaptive Ferroptosis. ACS Nano. 2023;17:13792–810.

    Article  CAS  PubMed  Google Scholar 

  29. Kobayashi T, Kishigami S, Sone M, Inokuchi H, Mogi T, Ito K. Respiratory chain is required to maintain oxidized states of the DsbA-DsbB disulfide bond formation system in aerobically growing Escherichia coli cells. Proc Natl Acad Sci USA. 1997;94:11857–62.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  30. Sriramajayam K, Peng D, Lu H, Zhou S, Bhat N, McDonald OG, et al. Activation of NRF2 by APE1/REF1 is redox-dependent in Barrett’s related esophageal adenocarcinoma cells. Redox Biol. 2021;43:101970.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  31. Tang D, Chen X, Kang R, Kroemer G. Ferroptosis: molecular mechanisms and health implications. Cell Res. 2021;31:107–25.

    Article  CAS  PubMed  Google Scholar 

  32. Habib E, Linher-Melville K, Lin HX, Singh G. Expression of xCT and activity of system xc(-) are regulated by NRF2 in human breast cancer cells in response to oxidative stress. Redox Biol. 2015;5:33–42.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  33. Yuan S, Wei C, Liu G, Zhang L, Li J, Li L, et al. Sorafenib attenuates liver fibrosis by triggering hepatic stellate cell ferroptosis via HIF-1alpha/SLC7A11 pathway. Cell Prolif. 2022;55:e13158.

    Article  CAS  PubMed  Google Scholar 

  34. Feng H, Liu Y, Gan Y, Li M, Liu R, Liang Z, et al. AdipoR1 Regulates Ionizing Radiation-Induced Ferroptosis in HCC cells through Nrf2/xCT Pathway. Oxid Med Cell Longev. 2022;2022:8091464.

    Article  PubMed  PubMed Central  Google Scholar 

  35. Hayes JD, Dinkova-Kostova AT. The Nrf2 regulatory network provides an interface between redox and intermediary metabolism. Trends Biochem Sci. 2014;39:199–218.

    Article  CAS  PubMed  Google Scholar 

  36. Rada P, Rojo AI, Evrard-Todeschi N, Innamorato NG, Cotte A, Jaworski T, et al. Structural and functional characterization of Nrf2 degradation by the glycogen synthase kinase 3/beta-TrCP axis. Mol Cell Biol. 2012;32:3486–99.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  37. Sharma M, Chuang WW, Sun Z. Phosphatidylinositol 3-kinase/Akt stimulates androgen pathway through GSK3beta inhibition and nuclear beta-catenin accumulation. J Biol Chem. 2002;277:30935–41.

    Article  CAS  PubMed  Google Scholar 

  38. Murata H, Ihara Y, Nakamura H, Yodoi J, Sumikawa K, Kondo T. Glutaredoxin exerts an antiapoptotic effect by regulating the redox state of Akt. J Biol Chem. 2003;278:50226–33.

    Article  CAS  PubMed  Google Scholar 

  39. Yao F, Deng Y, Zhao Y, Mei Y, Zhang Y, Liu X, et al. A targetable LIFR-NF-kappaB-LCN2 axis controls liver tumorigenesis and vulnerability to ferroptosis. Nat Commun. 2021;12:7333.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  40. Gao R, Kalathur RKR, Coto-Llerena M, Ercan C, Buechel D, Shuang S, et al. YAP/TAZ and ATF4 drive resistance to Sorafenib in hepatocellular carcinoma by preventing ferroptosis. EMBO Mol Med. 2021;13:e14351.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  41. Lei G, Zhang Y, Koppula P, Liu X, Zhang J, Lin SH, et al. The role of ferroptosis in ionizing radiation-induced cell death and tumor suppression. Cell Res. 2020;30:146–62.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  42. Viswanathan VS, Ryan MJ, Dhruv HD, Gill S, Eichhoff OM, Seashore-Ludlow B, et al. Dependency of a therapy-resistant state of cancer cells on a lipid peroxidase pathway. Nature. 2017;547:453–7.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  43. Wu J, Minikes AM, Gao M, Bian H, Li Y, Stockwell BR, et al. Intercellular interaction dictates cancer cell ferroptosis via NF2-YAP signalling. Nature. 2019;572:402–6.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  44. Gao M, Yi J, Zhu J, Minikes AM, Monian P, Thompson CB, et al. Role of Mitochondria in Ferroptosis. Mol Cell. 2019;73:354–63.e353.

    Article  CAS  PubMed  Google Scholar 

  45. Ma X, Dang C, Min W, Diao Y, Hui W, Wang X, et al. Downregulation of APE1 potentiates breast cancer cells to olaparib by inhibiting PARP-1 expression. Breast Cancer Res Treat. 2019;176:109–17.

    Article  CAS  PubMed  Google Scholar 

  46. Bersuker K, Hendricks JM, Li Z, Magtanong L, Ford B, Tang PH, et al. The CoQ oxidoreductase FSP1 acts parallel to GPX4 to inhibit ferroptosis. Nature. 2019;575:688–92.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  47. Doll S, Proneth B, Tyurina YY, Panzilius E, Kobayashi S, Ingold I, et al. ACSL4 dictates ferroptosis sensitivity by shaping cellular lipid composition. Nat Chem Biol. 2017;13:91–8.

    Article  CAS  PubMed  Google Scholar 

  48. Liu J, Ren Z, Yang L, Zhu L, Li Y, Bie C, et al. The NSUN5-FTH1/FTL pathway mediates ferroptosis in bone marrow-derived mesenchymal stem cells. Cell Death Discov. 2022;8:99.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  49. Dodson M, Castro-Portuguez R, Zhang DD. NRF2 plays a critical role in mitigating lipid peroxidation and ferroptosis. Redox Biol. 2019;23:101107.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  50. Cuadrado A. Structural and functional characterization of Nrf2 degradation by glycogen synthase kinase 3/beta-TrCP. Free Radic Biol Med. 2015;88:147–57.

    Article  CAS  PubMed  Google Scholar 

  51. Park MS, Kim CS, Joo HK, Lee YR, Kang G, Kim SJ, et al. Cytoplasmic localization and redox cysteine residue of APE1/Ref-1 are associated with its anti-inflammatory activity in cultured endothelial cells. Mol Cells. 2013;36:439–45.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  52. Di Maso V, Avellini C, Croce LS, Rosso N, Quadrifoglio F, Cesaratto L, et al. Subcellular localization of APE1/Ref-1 in human hepatocellular carcinoma: possible prognostic significance. Mol Med. 2007;13:89–96.

    Article  PubMed  PubMed Central  Google Scholar 

  53. Lu GS, Li M, Xu CX, Wang D. APE1 stimulates EGFR-TKI resistance by activating Akt signaling through a redox-dependent mechanism in lung adenocarcinoma. Cell Death Dis. 2018;9:1111.

    Article  PubMed  PubMed Central  Google Scholar 

  54. Whitaker AM, Freudenthal BD. APE1: A skilled nucleic acid surgeon. DNA Repair. 2018;71:93–100.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  55. Li M, Yang X, Lu X, Dai N, Zhang S, Cheng Y, et al. APE1 deficiency promotes cellular senescence and premature aging features. Nucleic Acids Res. 2018;46:5664–77.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  56. Liu J, Kang R, Tang D. Signaling pathways and defense mechanisms of ferroptosis. FEBS J. 2022;289:7038–50.

    Article  CAS  PubMed  Google Scholar 

  57. Chen PH, Tseng WH, Chi JT. The Intersection of DNA Damage Response and Ferroptosis-A Rationale for Combination Therapeutics. Biology. 2020;9:187.

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Funding

This work was supported by the National Natural Science Foundation of China (32171407 and 82373183) and the Priority Academic Program Development of Jiangsu Higher Education Institutions.

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  1. These authors contributed equally: Yu Du, Yu Zhou.

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  1. Jiangsu Key Laboratory for Molecular and Medical Biotechnology, College of Life Sciences, Nanjing Normal University, 1 WenYuan Road, Nanjing, 210023, China

    Yu Du, Yu Zhou, Xinyu Yan, Feiyan Pan, Lingfeng He, Zhigang Guo & Zhigang Hu

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Contributions

Y.D., Y.Z., and X.Y. performed study concept and design; Z.H., Y.D., Z.G., L.H. and F.P. performed development of methodology and writing, review and revision of the paper; Y.D., Y.Z., X.Y., Z.H. and Z.G. provided acquisition, analysis and interpretation of data, and statistical analysis; F.P., Z.H., L.H. and Z.G. provided technical and material support. All authors read and approved the final paper.

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Correspondence to Zhigang Guo or Zhigang Hu.

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This research was approved by the Ethics Committee of Nanjing Normal University. All animal experiments were performed according to procedures approved by the Laboratory Animal Care Committee at Nanjing Normal University, in accordance with the National Institutes of Health Guide for the Care and Use of Laboratory Animals.

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Du, Y., Zhou, Y., Yan, X. et al. APE1 inhibition enhances ferroptotic cell death and contributes to hepatocellular carcinoma therapy. Cell Death Differ 31, 431–446 (2024). https://doi.org/10.1038/s41418-024-01270-0

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