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Phenazine derivatives attenuate the stemness of breast cancer cells through triggering ferroptosis

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Abstract

Breast cancer stem cells (BCSCs) are positively correlated with the metastasis, chemoresistance, and recurrence of breast cancer. However, there are still no drugs targeting BCSCs in clinical using for breast cancer treatment. Here, we tried to screen out small-molecule compounds targeting BCSCs from the phenazine library established by us before. We focused on the compounds without affecting cell viability and screened out three potential compounds (CPUL119, CPUL129, CPUL149) that can significantly attenuate the stemness of breast cancer cells, as evident by the decrease of stemness marker expression, CD44+/CD24– subpopulation, mammary spheroid-formation ability, and tumor-initiating capacity. Additionally, these compounds suppressed the metastatic ability of breast cancer cells in vitro and in vivo. Combined with the transcriptome sequencing analysis, ferroptosis was shown on the top of the most upregulated pathways by CPUL119, CPUL129, and CPUL149, respectively. Mechanistically, we found that these three compounds could trigger ferroptosis by accumulating and sequestering iron in lysosomes through interacting with iron, and by regulating the expression of proteins (IRP2, TfR1, ferritin) engaged in iron transport and storage. Furthermore, inhibition of ferroptosis rescued the suppression of these three compounds on breast cancer cell stemness. This study suggests that CPUL119, CPUL129, and CPUL149 can specifically inhibit the stemness of breast cancer cells through triggering ferroptosis and may be the potential compounds for breast cancer treatment.

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Abbreviations

BCSCs:

Breast cancer stem cells

CSCs:

Cancer stem cells

LSCs:

Leukemia stem cells

FDA:

Food and Drug Administration

ROS:

Reactive oxygen species

IRP2:

Iron-responsive element-binding protein 2

TfR1:

Transferrin receptor 1

IRE:

Iron regulatory element

FPN:

Ferroportin

GSEA:

Gene Set Enrichment Analysis

EMT:

Epithelial–mesenchymal transition

Taxol:

Paclitaxel

Adr:

Adriamycin

GSH:

Glutathione

FAC:

Ferric ammonium citrate

NMR:

Nuclear Magnetic Resonance

References

  1. Chen W, Qin Y, Liu S (2018) Cytokines, breast cancer stem cells (BCSCs) and chemoresistance. Clin Transl Med 7(1):27

    Article  PubMed  PubMed Central  Google Scholar 

  2. Fiorentino S, Urueña C, Lasso P, Prieto K, Barreto A (2020) Phyto-immunotherapy, a complementary therapeutic option to decrease metastasis and attack breast cancer stem cells. Front Oncol 10:1334

    Article  PubMed  PubMed Central  Google Scholar 

  3. Peitzsch C, Tyutyunnykova A, Pantel K, Dubrovska A (2017) Cancer stem cells: the root of tumor recurrence and metastases. Semin Cancer Biol 44:10–24

    Article  CAS  PubMed  Google Scholar 

  4. Yang F, Xu J, Tang L, Guan X (2017) Breast cancer stem cell: the roles and therapeutic implications. Cell Mol Life Sci 74(6):951–966

    Article  CAS  PubMed  Google Scholar 

  5. Yang Y, Li X, Wang T, Guo Q, Xi T, Zheng L (2020) Emerging agents that target signaling pathways in cancer stem cells. J Hematol Oncol 13(1):60

    Article  PubMed  PubMed Central  Google Scholar 

  6. Norsworthy KJ, By K, Subramaniam S, Zhuang L, Del Valle PL, Przepiorka D, Shen YL, Sheth CM, Liu C, Leong R et al (2019) FDA approval summary: Glasdegib for newly diagnosed acute myeloid Leukemia. Clin Cancer Res 25(20):6021–6025

    Article  CAS  PubMed  Google Scholar 

  7. Beziat G, Ysebaert L (2020) Tagraxofusp for the treatment of Blastic Plasmacytoid Dendritic Cell Neoplasm (BPDCN): a brief report on emerging data. Onco Targets Ther 13:5199–5205

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Raggi C, Gammella E, Correnti M, Buratti P, Forti E, Andersen JB, Alpini G, Glaser S, Alvaro D, Invernizzi P et al (2017) Dysregulation of iron metabolism in Cholangiocarcinoma stem-like cells. Sci Rep 7(1):17667

    Article  PubMed  PubMed Central  Google Scholar 

  9. Ozer U (2016) The role of Iron on breast cancer stem-like cells. Cell Mol Biol (Noisy-le-grand) 62(4):25–30

    CAS  Google Scholar 

  10. Basuli D, Tesfay L, Deng Z, Paul B, Yamamoto Y, Ning G, Xian W, McKeon F, Lynch M, Crum CP et al (2017) Iron addiction: a novel therapeutic target in ovarian cancer. Oncogene 36(29):4089–4099

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. Bajbouj K, Shafarin J, Hamad M (2019) Estrogen-dependent disruption of intracellular iron metabolism augments the cytotoxic effects of doxorubicin in select breast and ovarian cancer cells. Cancer Manage Res 11:4655–4668

    Article  CAS  Google Scholar 

  12. Ma S, Henson ES, Chen Y, Gibson SB (2016) Ferroptosis is induced following siramesine and lapatinib treatment of breast cancer cells. Cell Death Dis 7(7):e2307

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Chang VC, Cotterchio M, Khoo E (2019) Iron intake, body iron status, and risk of breast cancer: a systematic review and meta-analysis. BMC Cancer 19(1):543

    Article  PubMed  PubMed Central  Google Scholar 

  14. Ma S, Dielschneider RF, Henson ES, Xiao W, Gibson SB (2017) Ferroptosis and autophagy induced cell death occur independently after siramesine and lapatinib treatment in breast cancer cells. PLoS One 12(8):e0182921

    Article  PubMed  PubMed Central  Google Scholar 

  15. Plays M, Müller S, Rodriguez R (2021) Chemistry and biology of ferritin. Metallomics 13(5):mfab021

    Article  PubMed  Google Scholar 

  16. Torii S, Shintoku R, Kubota C, Yaegashi M, Torii R, Sasaki M, Suzuki T, Mori M, Yoshimoto Y, Takeuchi T et al (2016) An essential role for functional lysosomes in ferroptosis of cancer cells. Biochem J 473(6):769–777

    Article  CAS  PubMed  Google Scholar 

  17. Mai TT, Hamai A, Hienzsch A, Caneque T, Muller S, Wicinski J, Cabaud O, Leroy C, David A, Acevedo V et al (2017) Salinomycin kills cancer stem cells by sequestering iron in lysosomes. Nat Chem 9(10):1025–1033

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. Corte-Rodríguez M, Blanco-González E, Bettmer J, Montes-Bayón M (2019) Quantitative analysis of transferrin receptor 1 (TfR1) in individual breast cancer cells by means of labeled antibodies and elemental (ICP-MS) detection. Anal Chem 91(24):15532–15538

    Article  PubMed  Google Scholar 

  19. Xu Y, Wang Q, Li X, Chen Y, Xu G (2021) Itraconazole attenuates the stemness of nasopharyngeal carcinoma cells via triggering ferroptosis. Environ Toxicol 36(2):257–266

    Article  CAS  PubMed  Google Scholar 

  20. Sun J, Cheng X, Pan S, Wang L, Dou W, Liu J, Shi X (2020) Dichloroacetate attenuates the stemness of colorectal cancer cells via trigerring ferroptosis through sequestering iron in lysosomes. Environ Toxicol 36(4):520–529

    Article  PubMed  Google Scholar 

  21. Miller LD, Coffman LG, Chou JW, Black MA, Bergh J, D’Agostino R Jr, Torti SV, Torti FM (2011) An iron regulatory gene signature predicts outcome in breast cancer. Cancer Res 71(21):6728–6737

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  22. Müller S, Sindikubwabo F, Cañeque T, Lafon A, Versini A, Lombard B, Loew D, Wu TD, Ginestier C, Charafe-Jauffret E et al (2020) CD44 regulates epigenetic plasticity by mediating iron endocytosis. Nat Chem 12(10):929–938

    Article  PubMed  PubMed Central  Google Scholar 

  23. Wang K, Chen X, Zuyi W, Chen L, Fu W (2021) Lysosome Fe(2+) release is responsible for etoposide- and cisplatin-induced stemness of small cell lung cancer cells. Environ Toxicol 36(8):1654–1663

    Article  CAS  PubMed  Google Scholar 

  24. Zhao B, Li X, Wang Y, Shang P (2018) Iron-dependent cell death as executioner of cancer stem cells. J Exp Clin Cancer Res 37(1):79

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  25. Ma S, Dielschneider RF, Henson ES, Xiao W, Choquette TR, Blankstein AR, Chen Y, Gibson SB (2017) Ferroptosis and autophagy induced cell death occur independently after siramesine and lapatinib treatment in breast cancer cells. PLoS One 12(8):e0182921

    Article  PubMed  PubMed Central  Google Scholar 

  26. Mulkearns-Hubert EE, Torre-Healy LA, Silver DJ, Eurich JT, Bayik D, Serbinowski E, Hitomi M, Zhou J, Przychodzen B, Zhang R et al (2019) Development of a Cx46 targeting strategy for cancer stem cells. Cell Rep 27(4):1062–1072 (e1065)

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  27. Kumar H, Chattopadhyay S, Das N, Shree S, Patel D, Mohapatra J, Gurjar A, Kushwaha S, Singh AK, Dubey S et al (2020) Leprosy drug clofazimine activates peroxisome proliferator-activated receptor-γ and synergizes with imatinib to inhibit chronic myeloid leukemia cells. Haematologica 105(4):971–986

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  28. Durusu İZ, Hüsnügil HH, Ataş H, Biber A, Gerekçi S, Güleç EA, Özen C (2017) Anti-cancer effect of clofazimine as a single agent and in combination with cisplatin on U266 multiple myeloma cell line. Leuk Res 55:33–40

    Article  CAS  PubMed  Google Scholar 

  29. Ahmed K, Koval A, Xu J, Bodmer A, Katanaev VL (2019) Towards the first targeted therapy for triple-negative breast cancer: repositioning of clofazimine as a chemotherapy-compatible selective Wnt pathway inhibitor. Cancer Lett 449:45–55

    Article  CAS  PubMed  Google Scholar 

  30. Lu Y, Wang L, Xiaobing X, Tao L, Jianmin Z (2017) Design, combinatorial synthesis and biological evaluations of novel 3-amino-1′-((1-aryl-1H-1,2,3-triazol-5-yl)methyl)-2′-oxospiro[benzo[a] pyrano[2,3-c]phenazine-1,3′-indoline]-2-carbonitrile antitumor hybrid molecules. Eur J Med Chem 135:125–141

    Article  CAS  PubMed  Google Scholar 

  31. Mei-Chen Z, Shu-Hui G, Guang-Pan L, Chen-Cheng L, Han-Mei X (2019) Facile synthesis and cytotoxicity of phenazine-chromene hybrid molecules derived from phenazine natural product. Combin Chem High Throughput Screen 22(1):35–40

    Article  Google Scholar 

  32. Lu Y, Yan Y, Wang L, Wang X, Gao J, Xi T, Wang Z, Jiang F (2016) Design, facile synthesis and biological evaluations of novel pyrano[3,2-a]phenazine hybrid molecules as antitumor agents. Eur J Med Chem 127:928–943

    Article  PubMed  Google Scholar 

  33. Gao J, Chen M, Tong X, Zhu H, Yan H, Liu D, Li W, Qi S, Xiao D, Wang Y (2015) Synthesis, antitumor activity, and structure-activity relationship of some benzo[a]pyrano[2,3-c]phenazine derivatives. Combin Chem High Throughput Screen 18(10):960–974

    Article  CAS  Google Scholar 

  34. Ghafouri-Fard S, Taheri M (2019) UCA1 long non-coding RNA: an update on its roles in malignant behavior of cancers. Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie 120:109459

    Article  CAS  Google Scholar 

  35. Klinge CM (2018) Non-coding RNAs in breast cancer: intracellular and intercellular communication. Non-coding RNA 4(4):40

    Article  CAS  PubMed Central  Google Scholar 

  36. Gao M, Yi J, Zhu J, Minikes AM, Monian P, Thompson CB, Jiang X (2019) Role of Mitochondria in Ferroptosis. Mol Cell 73(2):354-363.e353

    Article  CAS  PubMed  Google Scholar 

  37. Eun K, Ham SW, Kim H (2017) Cancer stem cell heterogeneity: origin and new perspectives on CSC targeting. BMB Rep 50(3):117–125

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  38. Wang T, Shigdar S, Gantier MP, Hou Y, Wang L, Li Y, Shamaileh HA, Yin W, Zhou SF, Zhao X et al (2015) Cancer stem cell targeted therapy: progress amid controversies. Oncotarget 6(42):44191–44206

    Article  PubMed  PubMed Central  Google Scholar 

  39. O’Conor CJ, Chen T, González I, Cao D, Peng Y (2018) Cancer stem cells in triple-negative breast cancer: a potential target and prognostic marker. Biomark Med 12(7):813–820

    Article  CAS  PubMed  Google Scholar 

  40. Kim SE, Zhang L, Ma K, Riegman M, Chen F, Ingold I, Conrad M, Turker MZ, Gao M, Jiang X et al (2016) Ultrasmall nanoparticles induce ferroptosis in nutrient-deprived cancer cells and suppress tumour growth. Nat Nanotechnol 11(11):977–985

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  41. Versini A, Colombeau L, Hienzsch A, Gaillet C, Retailleau P, Debieu S, Muller S, Caneque T, Rodriguez R (2020) Salinomycin derivatives kill breast cancer stem cells via Lysosomal iron targeting. Chemistry 26(33):7416–7424

    Article  CAS  PubMed  Google Scholar 

  42. Laraia L, Garivet G, Foley DJ, Kaiser N, Müller S, Zinken S, Pinkert T, Wilke J, Corkery D, Pahl A et al (2020) Image-based morphological profiling identifies a Lysosomotropic, iron-sequestering autophagy inhibitor. Angew Chem Int Ed Engl 59(14):5721–5729

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  43. Kurz T, Terman A, Gustafsson B, Brunk UT (2008) Lysosomes in iron metabolism, ageing and apoptosis. Histochem Cell Biol 129(4):389–406

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  44. Goncalves J, Moog S, Morin A, Gentric G, Müller S, Morrell AP, Kluckova K, Stewart TJ, Andoniadou CL, Lussey-Lepoutre C et al (2021) Loss of SDHB promotes dysregulated iron homeostasis, oxidative stress, and sensitivity to ascorbate. Cancer Res 81(13):3480–3494

    Article  CAS  PubMed  Google Scholar 

  45. Rycaj K, Tang DG (2015) Cell-of-origin of cancer versus cancer stem cells: assays and interpretations. Cancer Res 75(19):4003–4011

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  46. Su S, Chen J, Yao H, Liu J, Yu S, Lao L, Wang M, Luo M, Xing Y, Chen F et al (2018) CD10(+)GPR77(+) cancer-associated fibroblasts promote cancer formation and chemoresistance by sustaining cancer stemness. Cell 172(4):841-856.e816

    Article  CAS  PubMed  Google Scholar 

  47. Schonberg DL, Miller TE, Wu Q, Flavahan WA, Das NK, Hale JS, Hubert CG, Mack SC, Jarrar AM, Karl RT et al (2015) Preferential iron trafficking characterizes glioblastoma stem-like cells. Cancer Cell 28(4):441–455

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  48. Raghupathy R, Manwani D, Little JA (2010) Iron overload in sickle cell disease. Adv Hematol 2010:272940

    Article  PubMed  PubMed Central  Google Scholar 

  49. Pietrangelo A (2016) Iron and the liver. Liver Int 36(Suppl 1):116–123

    Article  CAS  PubMed  Google Scholar 

  50. Kremastinos DT, Farmakis D (2011) Iron overload cardiomyopathy in clinical practice. Circulation 124(20):2253–2263

    Article  PubMed  Google Scholar 

  51. Zhao W, Liu J, Li Y, Chen Z, Qi D, Zhang Z (2021) Immune effect of active components of traditional Chinese medicine on triple-negative breast cancer. Front Pharmacol 12:731741

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  52. Tharmapalan P, Mahendralingam M, Berman HK, Khokha R (2019) Mammary stem cells and progenitors: targeting the roots of breast cancer for prevention. EMBO J 38(14):e100852

    Article  PubMed  PubMed Central  Google Scholar 

  53. Guo Q, Wang T, Yang Y, Gao L, Zhao Q, Zhang W, Xi T, Zheng L (2020) Transcriptional Factor Yin Yang 1 promotes the stemness of breast cancer cells by suppressing miR-873-5p transcriptional activity. Mol Ther Nucleic acids 21:527–541

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  54. Garrison AT, Abouelhassan Y, Kallifidas D, Tan H, Kim YS, Jin S, Luesch H, Huigens RW 3rd (2018) An efficient buchwald-hartwig/reductive cyclization for the scaffold diversification of halogenated phenazines: potent antibacterial targeting, biofilm eradication, and prodrug exploration. J Med Chem 61(9):3962–3983

    Article  CAS  PubMed  Google Scholar 

  55. Yang H, Kundra S, Chojnacki M, Liu K, Fuse MA, Abouelhassan Y, Kallifidas D, Zhang P, Huang G, Jin S et al (2021) A modular synthetic route involving N-aryl-2-nitrosoaniline intermediates leads to a new series of 3-substituted halogenated phenazine antibacterial agents. J Med Chem 64(11):7275–7295

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  56. Yang J, Li T, Gao C, Lv X, Liu K, Song H, Xing Y, Xi T (2014) FOXO1 3’UTR functions as a ceRNA in repressing the metastases of breast cancer cells via regulating miRNA activity. FEBS Lett 588(17):3218–3224

    Article  CAS  PubMed  Google Scholar 

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Funding

This work was supported by the Project Program of National Nature Science Foundation of China (Grant No. 82173842, 81872757), Nature Science Foundation of Jiangsu Province of China (Grant No. BK20201329), the Fundamental Research Funds for the Central Universities (grant No. 2632018ZD01, No. 2632020ZD10), Innovation and Entrepreneurship Training Program for Undergraduate (No. 201910316220, No. 202010316245), the Medical Science and Technology Research Project of Henan Province (no. SBGJ202003010), the Science and Technology Research Project of Henan Province (no. 202102310158) and the Priority Academic Program Development (PAPD) of Jiangsu Higher Education Institutions.

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Author notes

  1. Yue Yang, Yuanyuan Lu, and Chunhua Zhang contributed equally to this work.

Authors and Affiliations

  1. School of Life Science and Technology, School of Engineering, Jiangsu Key Laboratory of Carcinogenesis and Intervention, China Pharmaceutical University, 639 Longmian Road, Nanjing, 211198, People’s Republic of China

    Yue Yang, Yuanyuan Lu, Chunhua Zhang, Ting Wang, Zhuolu Xia, Jing Liu, Xiangyu Cheng, Tao Xi, Feng Jiang & Lufeng Zheng

  2. Department of Pharmacy, Affiliated Cancer Hospital of Zhengzhou University, Henan Cancer Hospital, 127 Dongming Road, Zhengzhou, 450003, People’s Republic of China

    Qianqian Guo & Wenzhou Zhang

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  1. Yue Yang
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  2. Yuanyuan Lu
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  3. Chunhua Zhang
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  4. Qianqian Guo
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  10. Tao Xi
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  11. Feng Jiang
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Contributions

LZ, FJ and TX designed the research. YL and FJ approved the compounds. YY, CZ, ZX, JL, XC, TW, QG analyzed the data. YY, CZ performed the research. YY and LZwrote the paper. All authors read and approved the final manuscript.

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Correspondence to Tao Xi, Feng Jiang or Lufeng Zheng.

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All the experiments were obtained the approval of the Ethics Committee for Animal Experimentation of China Pharmaceutical University.

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Yang, Y., Lu, Y., Zhang, C. et al. Phenazine derivatives attenuate the stemness of breast cancer cells through triggering ferroptosis. Cell. Mol. Life Sci. 79, 360 (2022). https://doi.org/10.1007/s00018-022-04384-1

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