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Dysregulation of iron homeostasis by TfR-1 renders EZH2 wild type diffuse large B-cell lymphoma resistance to EZH2 inhibition

Acta Pharmacologica Sinica volume 44, pages 2113–2124 (2023)Cite this article

Abstract

EZH2 has been regarded as an efficient target for diffuse large B-cell lymphoma (DLBCL), but the clinical benefits of EZH2 inhibitors (EZH2i) are limited. To date, only EPZ-6438 has been approved by FDA for the treatment of follicular lymphoma and epithelioid sarcoma. We have discovered a novel EZH1/2 inhibitor HH2853 with a better antitumor effect than EPZ-6438 in preclinical studies. In this study we explored the molecular mechanism underlying the primary resistance to EZH2 inhibitors and sought for combination therapy strategy to overcome it. By analyzing EPZ-6438 and HH2853 response profiling, we found that EZH2 inhibition increased intracellular iron through upregulation of transferrin receptor 1 (TfR-1), ultimately triggered resistance to EZH2i in DLBCL cells. We demonstrated that H3K27ac gain by EZH2i enhanced c-Myc transcription, which contributed to TfR-1 overexpression in insensitive U-2932 and WILL-2 cells. On the other hand, EZH2i impaired the occurrence of ferroptosis by upregulating the heat shock protein family A (Hsp70) member 5 (HSPA5) and stabilizing glutathione peroxidase 4 (GPX4), a ferroptosis suppressor; co-treatment with ferroptosis inducer erastin effectively overrode the resistance of DLBCL to EZH2i in vitro and in vivo. Altogether, this study reveals iron-dependent resistance evoked by EZH2i in DLBCL cells, and suggests that combination with ferroptosis inducer may be a promising therapeutic strategy.

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Fig. 1: HH2853 displays different responses in DLBCL in vitro and in vivo.
Fig. 2: EZH2 inhibition affects intracellular LIP content through TfR-1.
Fig. 3: EZH2i affects the expression of TfR-1 by regulating c-Myc.
Fig. 4: EZH2i stabilizes GPX4 through HSPA5 and inhibits cell ferroptosis.
Fig. 5: Combination of EZH2i and Erastin induced ferroptosis of drug-resistant cells.
Fig. 6: EZH2i and Erastin synergistically inhibit DLBCL xenografts growth.

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References

  1. Di Croce L, Helin K. Transcriptional regulation by Polycomb group proteins. Nat Struct Mol Biol. 2013;20:1147–55.

    Article  PubMed  Google Scholar 

  2. Duan R, Du W, Guo W. EZH2: a novel target for cancer treatment. J Hematol Oncol. 2020;13:104.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. Eich ML, Athar M, Ferguson JE 3rd, Varambally S. EZH2-targeted therapies in cancer: hype or a reality. Cancer Res. 2020;80:5449–58.

    Article  PubMed  PubMed Central  Google Scholar 

  4. Knutson SK, Kawano S, Minoshima Y, Warholic NM, Huang KC, Xiao Y, et al. Selective Inhibition of EZH2 by EPZ-6438 leads to potent antitumor activity in EZH2-mutant non-Hodgkin lymphoma. Mol Cancer Ther. 2014;13:842.

    Article  CAS  PubMed  Google Scholar 

  5. Morschhauser F, Tilly H, Chaidos A, McKay P, Phillips T, Assouline S, et al. Tazemetostat for patients with relapsed or refractory follicular lymphoma: an open-label, single-arm, multicentre, phase 2 trial. Lancet Oncol. 2020;21:1433–42.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Italiano A, Soria J-C, Toulmonde M, Michot JM, Lucchesi C, Varga A, et al. Tazemetostat, an EZH2 inhibitor, in relapsed or refractory B-cell non-Hodgkin lymphoma and advanced solid tumours: a first-in-human, open-label, phase 1 study. The Lancet Oncology. 2018;19:649–59.

  7. McCabe MT, Ott HM, Ganji G, Korenchuk S, Thompson C, Van Aller GS, et al. EZH2 inhibition as a therapeutic strategy for lymphoma with EZH2-activating mutations. Nature. 2012;492:108–12.

    Article  CAS  PubMed  Google Scholar 

  8. Torti SV, Torti FM. Iron and Cancer: 2020 Vision. Cancer Res. 2020;80:5435–48.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Wang Y, Yu L, Ding J, Chen Y. Iron metabolism in cancer. Int J Mol Sci. 2018;20:95.

    Article  PubMed  PubMed Central  Google Scholar 

  10. Wang YF, Zhang J, Su Y, Shen YY, Jiang DX, Hou YY, et al. G9a regulates breast cancer growth by modulating iron homeostasis through the repression of ferroxidase hephaestin. Nat Commun. 2017;8:274.

    Article  PubMed  PubMed Central  Google Scholar 

  11. Kazan HH, Urfali-Mamatoglu C, Gunduz U. Iron metabolism and drug resistance in cancer. Biometals. 2017;30:629–41.

    Article  CAS  PubMed  Google Scholar 

  12. Shen Z, Song J, Yung BC, Zhou Z, Wu A, Chen X. Emerging strategies of cancer therapy based on ferroptosis. Adv Mater. 2018;30:e1704007–e.

    Article  PubMed  PubMed Central  Google Scholar 

  13. 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 

  14. Hangauer MJ, Viswanathan VS, Ryan MJ, Bole D, Eaton JK, Matov A, et al. Drug-tolerant persister cancer cells are vulnerable to GPX4 inhibition. Nature. 2017;551:247–50.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Hassannia B, Vandenabeele P, Vanden Berghe T. Targeting ferroptosis to iron out cancer. Cancer Cell. 2019;35:830–49.

    Article  CAS  PubMed  Google Scholar 

  16. Chen XX, Shen QQ, Zhao Z, Fang YF, Yang JY, Gao YL, et al. Abstract 5436: HH2853 is a selective small molecular dual inhibitor of EZH1/2 with potent anti-tumor activities. Cancer Res. 2022;82:5436.

    Article  Google Scholar 

  17. Huang X, Yan J, Zhang M, Wang Y, Chen Y, Fu X, et al. Targeting epigenetic crosstalk as a therapeutic strategy for EZH2-aberrant solid tumors. Cell. 2018;175:186–99.e19.

    Article  CAS  PubMed  Google Scholar 

  18. Liu Y, Gu Y, Han Y, Zhang Q, Jiang Z, Zhang X, et al. Tumor Exosomal RNAs Promote Lung Pre-metastatic Niche Formation by Activating Alveolar Epithelial TLR3 to Recruit Neutrophils. Cancer Cell. 2016;30:243–56.

  19. Morales M, Xue X. Targeting iron metabolism in cancer therapy. Theranostics. 2021;11:8412–29.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  20. Nemeth E, Ganz T. Hepcidin and iron in health and disease. Annu Rev Med. 2023;74:261–77.

    Article  PubMed  Google Scholar 

  21. Ganz T, Nemeth E. Hepcidin and iron homeostasis. Biochim Biophys Acta. 2012;1823:1434–43.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  22. Lu B, Chen XB, Ying MD, He QJ, Cao J, Yang B. The role of ferroptosis in cancer development and treatment response. Front Pharmacol. 2018;8:992.

    Article  PubMed  PubMed Central  Google Scholar 

  23. Yang H, Said AM, Huang H, Papa APD, Jin G, Wu S, et al. Chlorogenic acid depresses cellular bioenergetics to suppress pancreatic carcinoma through modulating c-Myc-TFR1 axis. Phytother Res. 2021;35:2200–10.

    Article  CAS  PubMed  Google Scholar 

  24. O’Donnell KA, Yu D, Zeller KI, Kim JW, Racke F, Thomas-Tikhonenko A, et al. Activation of transferrin receptor 1 by c-Myc enhances cellular proliferation and tumorigenesis. Mol Cell Biol. 2006;26:2373–86.

    Article  PubMed  PubMed Central  Google Scholar 

  25. Lu Y, Yang Q, Su Y, Ji Y, Li G, Yang X, et al. MYCN mediates TFRC-dependent ferroptosis and reveals vulnerabilities in neuroblastoma. Cell Death Dis. 2021;12:511.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  26. 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 

  27. Ooko E, Saeed ME, Kadioglu O, Sarvi S, Colak M, Elmasaoudi K, et al. Artemisinin derivatives induce iron-dependent cell death (ferroptosis) in tumor cells. Phytomedicine. 2015;22:1045–54.

    Article  CAS  PubMed  Google Scholar 

  28. Tang LJ, Zhou YJ, Xiong XM, Li NS, Zhang JJ, Luo XJ, et al. Ubiquitin-specific protease 7 promotes ferroptosis via activation of the p53/TfR1 pathway in the rat hearts after ischemia/reperfusion. Free Radic Biol Med. 2021;162:339–52.

    Article  CAS  PubMed  Google Scholar 

  29. 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 

  30. Bebber CM, Thomas ES, Stroh J, Chen Z, Androulidaki A, Schmitt A, et al. Ferroptosis response segregates small cell lung cancer (SCLC) neuroendocrine subtypes. Nat Commun. 2021;12:2048.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  31. 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 

  32. Mao C, Liu X, Zhang Y, Lei G, Yan Y, Lee H, et al. DHODH-mediated ferroptosis defence is a targetable vulnerability in cancer. Nature. 2021;593:586–90.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  33. Wang F, Min J. DHODH tangoing with GPX4 on the ferroptotic stage. Signal Transduct Target Ther. 2021;6:244.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  34. Zhu S, Zhang Q, Sun X, Zeh HJ 3rd, Lotze MT, Kang R, et al. HSPA5 regulates ferroptotic cell death in cancer cells. Cancer Res. 2017;77:2064–77.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  35. Zhao Y, Li Y, Zhang R, Wang F, Wang T, Jiao Y. The role of erastin in ferroptosis and its prospects in cancer therapy. Onco Targets Ther. 2020;13:5429–41.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  36. Lu Y, Zhao YL, Xiong M, Sun RJ, Cao XY, Wei ZJ, et al. Unmanipulated haploidentical donor and matched unrelated donor hematopoietic stem cell transplantation in patients with paroxysmal nocturnal hemoglobinuria: a single-center study. Leuk Lymphoma. 2022;63:1211–9.

    Article  CAS  PubMed  Google Scholar 

  37. Devin J, Cañeque T, Lin YL, Mondoulet L, Veyrune JL, Abouladze M, et al. Targeting cellular iron homeostasis with ironomycin in diffuse large B-cell lymphoma. Cancer Res. 2022;82:998–1012.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  38. Genta S, Pirosa MC, Stathis A. BET and EZH2 inhibitors: novel approaches for targeting cancer. Curr Oncol Rep. 2019;21:13.

    Article  PubMed  Google Scholar 

  39. Tremblay-LeMay R, Rastgoo N, Pourabdollah M, Chang H. EZH2 as a therapeutic target for multiple myeloma and other haematological malignancies. Biomark Res. 2018;6:34.

    Article  PubMed  PubMed Central  Google Scholar 

  40. Deb G, Singh AK, Gupta S. EZH2: not EZHY (easy) to deal. Mol Cancer Res. 2014;12:639–53.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  41. Sermer D, Pasqualucci L, Wendel HG, Melnick A, Younes A. Emerging epigenetic-modulating therapies in lymphoma. Nat Rev Clin Oncol. 2019;16:494–507.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  42. Honma D, Kanno O, Watanabe J, Kinoshita J, Hirasawa M, Nosaka E, et al. Novel orally bioavailable EZH1/2 dual inhibitors with greater antitumor efficacy than an EZH2 selective inhibitor. Cancer Sci. 2017;108:2069–78.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  43. Campisi A, Bonfanti R, Raciti G, Bonaventura G, Legnani L, Magro G, et al. Gene silencing of transferrin-1 receptor as a potential therapeutic target for human follicular and anaplastic thyroid cancer. Mol Ther Oncolytics. 2020;16:197–206.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  44. Hagag AA, Badraia IM, Abdelmageed MM, Hablas NM, Hazzaa SME, Nosair NA. Prognostic value of transferrin receptor-1 (CD71) expression in acute lymphoblastic leukemia. Endocr Metab Immune Disord Drug Targets. 2018;18:610–7.

    Article  CAS  PubMed  Google Scholar 

  45. Callens C, Coulon S, Naudin J, Radford-Weiss I, Boissel N, Raffoux E, et al. Targeting iron homeostasis induces cellular differentiation and synergizes with differentiating agents in acute myeloid leukemia. J Exp Med. 2010;207:731–50.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  46. Barabas K, Faulk WP. Transferrin receptors associate with drug resistance in cancer cells. Biochem Biophys Res Commun. 1993;197:702–8.

    Article  CAS  PubMed  Google Scholar 

  47. Chitambar CR, Wereley JP. Resistance to the antitumor agent gallium nitrate in human leukemic cells is associated with decreased gallium/iron uptake, increased activity of iron regulatory protein-1, and decreased ferritin production. J Biol Chem. 1997;272:12151–7.

    Article  CAS  PubMed  Google Scholar 

  48. Bonnah RA, Hoelter J, Steeghs L, Enns CA, So M, Muckenthaler MU. Lipooligosaccharide-independent alteration of cellular homeostasis in Neisseria meningitidis-infected epithelial cells. Cell Microbiol. 2005;7:869–85.

    Article  CAS  PubMed  Google Scholar 

  49. Kenneth NS, Mudie S, Naron S, Rocha S. TfR1 interacts with the IKK complex and is involved in IKK-NF-κB signalling. Biochem J. 2013;449:275–84.

    Article  CAS  PubMed  Google Scholar 

  50. Weng J, Chen L, Liu H, Yang XP, Huang L. Ferroptosis markers predict the survival, immune infiltration, and ibrutinib resistance of diffuse large B cell lymphoma. Inflammation. 2022;45:1146–61.

    Article  CAS  PubMed  Google Scholar 

  51. Chen D, Chu B, Yang X, Liu Z, Jin Y, Kon N, et al. iPLA2β-mediated lipid detoxification controls p53-driven ferroptosis independent of GPX4. Nat Commun. 2021;12:3644.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  52. Yang WS, Stockwell BR. Ferroptosis: death by lipid peroxidation. Trends Cell Biol. 2016;26:165–76.

    Article  CAS  PubMed  Google Scholar 

  53. Mizuno H, Kitada K, Nakai K, Sarai A. PrognoScan: a new database for meta-analysis of the prognostic value of genes. BMC Med Genom. 2009;2:18.

    Article  Google Scholar 

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Acknowledgements

This work was supported by grants from Program of Shanghai Academic Research Leader (22XD1404400), the Strategic Priority Research Program of the Chinese Academy of Sciences (XDA12020233), Youth Innovation Promotion Association of Chinese Academy of Sciences (2020281) and Lingang Laboratory (LG202103-02-05). We thank Haihe Biopharma Co., Ltd. for providing HH2853.

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Authors and Affiliations

  1. Division of Anti-Tumor Pharmacology, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai, 201203, China

    Lei Yu, Ya-fang Wang, Jian Xiao, Qian-qian Shen, Shuai-shuai Chi, Ying-lei Gao, Dong-ze Lin, Jian Ding, Yan-fen Fang & Yi Chen

  2. University of Chinese Academy of Sciences, No.19A Yuquan Road, Beijing, 100049, China

    Lei Yu, Jian Xiao, Shuai-shuai Chi, Jian Ding, Yan-fen Fang & Yi Chen

  3. State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai, 201203, China

    Jian Ding & Yan-fen Fang

  4. Shandong Laboratory of Yantai Drug Discovery, Bohai Rim Advanced Research Institute for Drug Discovery, Yantai, 264117, China

    Jian Ding & Yi Chen

  5. State Key Laboratory of Chemical Biology, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai, 201203, China

    Yi Chen

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Contributions

YC, YFF, JD, and LY designed the experiments. LY, YFW, JX, QQS, SSC, and DZL performed the research. LY and YFW analyzed the data, LY and JX wrote the paper which was revised by JD, YFF, and YC. All authors approved the final version of the paper.

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Correspondence to Jian Ding, Yan-fen Fang or Yi Chen.

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JD is the Chairman of Haihe Biopharma Co., Ltd. No competing interests is disclosed for rest of the authors.

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Yu, L., Wang, Yf., Xiao, J. et al. Dysregulation of iron homeostasis by TfR-1 renders EZH2 wild type diffuse large B-cell lymphoma resistance to EZH2 inhibition. Acta Pharmacol Sin 44, 2113–2124 (2023). https://doi.org/10.1038/s41401-023-01097-4

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