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Breast Cancer Research

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Open Access 01-12-2024 | Breast Cancer | Research

siRNA treatment targeting integrin α11 overexpressed via EZH2-driven axis inhibits drug-resistant breast cancer progression

Authors: Prakash Chaudhary, Kiran Yadav, Ho Jin Lee, Keon Wook Kang, Jongseo Mo, Jung-Ae Kim

Published in: Breast Cancer Research | Issue 1/2024

Abstract

Background

Breast cancer, the most prevalent cancer in women worldwide, faces treatment challenges due to drug resistance, posing a serious threat to patient survival. The present study aimed to identify the key molecules that drive drug resistance and aggressiveness in breast cancer cells and validate them as therapeutic targets.

Methods

Transcriptome microarray and analysis using PANTHER pathway and StemChecker were performed to identify the most significantly expressed genes in tamoxifen-resistant and adriamycin-resistant MCF-7 breast cancer cells. Clinical relevance of the key genes was determined using Kaplan-Meier survival analyses on The Cancer Genome Atlas dataset of breast cancer patients. Gene overexpression/knockdown, spheroid formation, flow cytometric analysis, chromatin immunoprecipitation, immunocytochemistry, wound healing/transwell migration assays, and cancer stem cell transcription factor activation profiling array were used to elucidate the regulatory mechanism of integrin α11 expression. Tumour-bearing xenograft models were used to demonstrate integrin α11 is a potential therapeutic target.

Results

Integrin α11 was consistently upregulated in drug-resistant breast cancer cells, and its silencing inhibited cancer stem cells (CSCs) and epithelial-mesenchymal transition (EMT) while restoring sensitivity to anticancer drugs. HIF1α, GLI-1, and EZH2 contributed the most to the regulation of integrin α11 and EZH2 expression, with EZH2 being more necessary for EZH2 autoinduction than HIF1α and GLI-1. Additionally, unlike HIF1α or EZH2, GLI-1 was the sole transcription factor activated by integrin-linked focal adhesion kinase, indicating GLI-1 as a key driver of the EZH2-integrin α11 axis operating for cancer stem cell survival and EMT. Kaplan-Meier survival analysis using The Cancer Genome Atlas (TCGA) dataset also revealed both EZH2 and integrin α11 could be strong prognostic factors of relapse-free and overall survival in breast cancer patients. However, the superior efficacy of integrin α11 siRNA therapy over EZH2 siRNA treatment was demonstrated by enhanced inhibition of tumour growth and prolonged survival in murine models bearing tumours.

Conclusion

Our findings elucidate that integrin α11 is upregulated by EZH2, forming a positive feedback circuit involving FAK-GLI-1 and contributing to drug resistance, cancer stem cell survival and EMT. Taken together, the results suggest integrin α11 as a promising prognostic marker and a powerful therapeutic target for drug-resistant breast cancer.

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Arnold M, Morgan E, Rumgay H, Mafra A, Singh D, Laversanne M, et al. Current and future burden of breast cancer: global statistics for 2020 and 2040. Breast. 2022;66:15–23.PubMedPubMedCentralCrossRef

Wilkinson L, Gathani T. Understanding breast cancer as a global health concern. Brit J Radiol. 2022;95(1130):20211033.PubMedCrossRef

Gautam J, Banskota S, Regmi SC, Ahn S, Jeon YH, Jeong H, et al. Tryptophan hydroxylase 1 and 5-HT7 receptor preferentially expressed in triple-negative breast cancer promote cancer progression through autocrine serotonin signaling. Mol Cancer. 2016;15(1):1–14.CrossRef

Goldberg J, Pastorello RG, Vallius T, Davis J, Cui YX, Agudo J, et al. The immunology of hormone receptor positive breast cancer. Front Immunol. 2021;12:674192.PubMedPubMedCentralCrossRef

Hanker AB, Sudhan DR, Arteaga CL. Overcoming endocrine resistance in breast cancer. Cancer Cell. 2020;37(4):496–513.PubMedPubMedCentralCrossRef

Gote V, Nookala AR, Bolla PK, Pal D. Drug resistance in metastatic breast cancer: tumor targeted nanomedicine to the rescue. Int J Mol Sci. 2021;22(9):4673.PubMedPubMedCentralCrossRef

Hernando C, Ortega-Morillo B, Tapia M, Moragón S, Martínez MT, Eroles P, et al. Oral selective estrogen receptor degraders (SERDs) as a novel breast cancer therapy: present and future from a clinical perspective. Int J Mol Sci. 2021;22(15):7812.PubMedPubMedCentralCrossRef

Patel HK, Bihani T. Selective estrogen receptor modulators (SERMs) and selective estrogen receptor degraders (SERDs) in cancer treatment. Pharmacol Ther. 2018;186:1–24.PubMedCrossRef

Group EBCTC. Effects of chemotherapy and hormonal therapy for early breast cancer on recurrence and 15-year survival: an overview of the randomised trials. Lancet. 2005;365(9472):1687–717.CrossRef

10.

Najafi S, Sadeghi M, Shajari MR, Abasvandi F, Mohebi K. The comparison of anthracycline-based and non-anthracycline-based regimens in adjuvant chemotherapy of HER2-positive non-metastatic breast cancers. Contemp Oncol (Pozn). 2018;22(2):108–12.PubMed

11.

Hurvitz SA, McAndrew NP, Bardia A, Press MF, Pegram M, Crown JP, et al. A careful reassessment of anthracycline use in curable breast cancer. NPJ Breast Cancer. 2021;7(1):134.PubMedPubMedCentralCrossRef

12.

Rampurwala MM, Rocque GB, Burkard ME. Update on adjuvant chemotherapy for early breast cancer. Breast Cancer: Basic Clin Res. 2014;8:BCBCR.

13.

Hosford SR, Miller TW. Clinical potential of novel therapeutic targets in breast cancer: CDK4/6, Src, JAK/STAT, PARP, HDAC, and PI3K/AKT/mTOR pathways. Pharmgenomics Pers Med. 2014: 203–15.

14.

Kumar R, Mandal M, Lipton A, Harvey H, Thompson CB. Overexpression of HER2 modulates bcl-2, bcl-XL, and tamoxifen-induced apoptosis in human MCF-7 breast cancer cells. Clin Cancer Res. 1996;2(7):1215–9.PubMed

15.

Rimawi MF, De Angelis C, Schiff R. Resistance to anti-HER2 therapies in breast cancer. Am Soc Clin Oncol Educ Book. 2015;35(1):e157–64.CrossRef

16.

Wu X, Yang H, Yu X, Qin J-J. Drug-resistant HER2-positive breast cancer: molecular mechanisms and overcoming strategies. Front Pharmacol. 2022;13:1012552.PubMedPubMedCentralCrossRef

17.

Rexer BN, Arteaga CL. Intrinsic and acquired resistance to HER2-targeted therapies in HER2 gene-amplified breast cancer: mechanisms and clinical implications. Crit Rev Oncog. 2012, 17(1).

18.

Cazet AS, Hui MN, Elsworth BL, Wu SZ, Roden D, Chan C-L, et al. Targeting stromal remodeling and cancer stem cell plasticity overcomes chemoresistance in triple-negative breast cancer. Nat Commun. 2018;9(1):2897.PubMedPubMedCentralCrossRef

19.

Barbato L, Bocchetti M, Di Biase A, Regad T. Cancer stem cells and targeting strategies. Cells. 2019;8(8):926.PubMedPubMedCentralCrossRef

20.

Seguin L, Desgrosellier JS, Weis SM, Cheresh DA. Integrins and cancer: regulators of cancer stemness, metastasis, and drug resistance. Trends Cell Biol. 2015;25(4):234–40.PubMedPubMedCentralCrossRef

21.

Cooper J, Giancotti FG. Integrin signaling in cancer: mechanotransduction, stemness, epithelial plasticity, and therapeutic resistance. Cancer Cell. 2019;35(3):347–67.PubMedPubMedCentralCrossRef

22.

Maugeri-Saccà M, Vigneri P, De Maria R. Cancer stem cells and chemosensitivity. Clin Cancer Res. 2011;17(15):4942–7.PubMedCrossRef

23.

Kotiyal S, Bhattacharya S. Breast cancer stem cells, EMT and therapeutic targets. Biochem Biophys Res Commun. 2014;453(1):112–6.PubMedCrossRef

24.

Giancotti FG, Tarone G. Positional control of cell fate through joint integrin/receptor protein kinase signaling. Annu Rev Cell Dev Biol. 2003;19(1):173–206.PubMedCrossRef

25.

Yousefi H, Vatanmakanian M, Mahdiannasser M, Mashouri L, Alahari NV, Monjezi MR, et al. Understanding the role of integrins in breast cancer invasion, metastasis, angiogenesis, and drug resistance. Oncogene. 2021;40(6):1043–63.PubMedCrossRef

26.

Zutter MM. Integrin-mediated adhesion: tipping the balance between chemosensitivity and chemoresistance. Adv Exp Med Biol. 2007: 87–100.

27.

Xiong J, Yan L, Zou C, Wang K, Chen M, Xu B, et al. Integrins regulate stemness in solid tumor: an emerging therapeutic target. J Hematol Oncol. 2021;14(1):1–18.CrossRef

28.

Goel HL, Pursell B, Chang C, Shaw LM, Mao J, Simin K, et al. GLI1 regulates a novel neuropilin-2/α6β1 integrin based autocrine pathway that contributes to breast cancer initiation. EMBO Mol Med. 2013;5(4):488–508.PubMedPubMedCentralCrossRef

29.

Campbell PS, Mavingire N, Khan S, Rowland LK, Wooten JV, Opoku-Agyeman A, et al. AhR ligand aminoflavone suppresses α6‐integrin–src–akt signaling to attenuate tamoxifen resistance in breast cancer cells. J Cell Physiol. 2019;234(1):108–21.CrossRef

30.

Adorno-Cruz V, Hoffmann AD, Liu X, Wray B, Keri RA, Liu H. ITGA2 is a target of miR-206 promoting cancer stemness and lung metastasis through enhanced ACLY and CCND1 expression in triple negative breast cancer. bioRxiv. 2019: 583062.

31.

Wang Z, Li Y, Xiao Y, Lin HP, Yang P, Humphries B, et al. Integrin α9 depletion promotes β-catenin degradation to suppress triple‐negative breast cancer tumor growth and metastasis. Int J Cancer. 2019;145(10):2767–80.PubMedPubMedCentralCrossRef

32.

Barnawi R, Al-Khaldi S, Colak D, Tulbah A, Al‐Tweigeri T, Fallatah M, et al. β1 integrin is essential for fascin‐mediated breast cancer stem cell function and disease progression. Int J Cancer. 2019;145(3):830–41.PubMedPubMedCentralCrossRef

33.

Zhang Y, Zhang Q, Cao Z, Huang Y, Cheng S, Pang D. HOXD3 plays a critical role in breast cancer stemness and drug resistance. Cell Physiol Biochem. 2018;46(4):1737–47.PubMedCrossRef

34.

Taddei I, Deugnier MA, Faraldo MM, Petit V, Bouvard D, Medina D, et al. β1 integrin deletion from the basal compartment of the mammary epithelium affects stem cells. Nat Cell Biol. 2008;10(6):716–22.PubMedPubMedCentralCrossRef

35.

Zhang L, Qu J, Qi Y, Duan Y, Huang Y-W, Zhou Z, et al. EZH2 engages TGFβ signaling to promote breast cancer bone metastasis via integrin β1-FAK activation. Nat Commun. 2022;13(1):2543.PubMedPubMedCentralCrossRef

36.

Dahal S, Chaudhary P, Kim J-A. Induction of promyelocytic leukemia zinc finger protein by miR-200c-3p restores sensitivity to anti-androgen therapy in androgen-refractory prostate cancer and inhibits the cancer progression via down-regulation of integrin α3β4. Cell Oncol. 2023: 1–14.

37.

Mi H, Ebert D, Muruganujan A, Mills C, Albou L-P, Mushayamaha T, et al. PANTHER version 16: a revised family classification, tree-based classification tool, enhancer regions and extensive API. Nucleic Acids Res. 2021;49(D1):D394–403.PubMedCrossRef

38.

Pinto JP, Kalathur RK, Oliveira DV, Barata T, Machado RS, Machado S, et al. StemChecker: a web-based tool to discover and explore stemness signatures in gene sets. Nucleic Acids Res. 2015;43(W1):W72–7.PubMedPubMedCentralCrossRef

39.

Vistica DT, Skehan P, Scudiero D, Monks A, Pittman A, Boyd MR. Tetrazolium-based assays for Cellular viability: a critical examination of selected parameters affecting Formazan production. Cancer Res. 1991;51(10):2515–20.PubMed

40.

Zhang Y, Wang M, Zhang X, Jiang Z, Zhang Y, Fu X et al. Helicid improves lipopolysaccharide-induced apoptosis of C6 cells by regulating SH2D5 DNA methylation via the CytC/Caspase9/Caspase3 signaling pathway. Contrast Media Mol I. 2022, 2022.

41.

Romaine A, Melleby AO, Alam J, Lobert VH, Lu N, Lockwood FE, et al. Integrin α11β1 and syndecan-4 dual receptor ablation attenuate cardiac hypertrophy in the pressure overloaded heart. Am J Physiol Heart Circ Physiol. 2022;322(6):H1057–71.PubMedCrossRef

42.

Shen B, Vardy K, Hughes P, Tasdogan A, Zhao Z, Yue R, et al. Integrin alpha11 is an osteolectin receptor and is required for the maintenance of adult skeletal bone mass. Elife. 2019;8:e42274.PubMedPubMedCentralCrossRef

43.

Nutt SL, Keenan C, Chopin M, Allan RS. EZH2 function in immune cell development. Biol Chem. 2020;401(8):933–43.PubMedCrossRef

44.

Cyrus S, Burkardt D, Weaver DD, Gibson WT. PRC2-complex related dysfunction in overgrowth syndromes: a review of EZH2, EED, and SUZ12 and their syndromic phenotypes. Am J Med Genet C Semin Med Genet. 2019. p. 519–31.

45.

Pastushenko I, Blanpain C. EMT transition states during tumor progression and metastasis. Trends Cell Biol. 2019;29(3):212–26.PubMedCrossRef

46.

Najafi M, Farhood B, Mortezaee K. Cancer stem cells (CSCs) in cancer progression and therapy. J Cell Physiol. 2019;234(6):8381–95.PubMedCrossRef

47.

Celià-Terrassa T, Jolly MK. Cancer stem cells and epithelial-to-mesenchymal transition in cancer metastasis. Cold Spring Harb Perspect Med. 2020;10(7):a036905.PubMedPubMedCentralCrossRef

48.

Ali S, Mondal N, Choudhry H, Rasool M, Pushparaj PN, Khan MA et al. Current management strategies in breast Cancer by targeting key altered molecular players. Front Oncol. 2016, 6.

49.

Hosford SR, Miller TW. Clinical potential of novel therapeutic targets in breast cancer: CDK4/6, Src, JAK/STAT, PARP, HDAC, and PI3K/AKT/mTOR pathways. Pharmgenomics Pers Med. 2014;7(null):203–15.PubMedPubMedCentral

50.

Droog M, Beelen K, Linn S, Zwart W. Tamoxifen resistance: from bench to bedside. Eur J Pharmacol. 2013;717(1):47–57.PubMedCrossRef

51.

Hernandez-Aya LF, Gonzalez-Angulo AM. Adjuvant systemic therapies in breast Cancer. Surg Clin North Am. 2013;93(2):473–91.PubMedCrossRef

52.

Berman AT, Thukral AD, Hwang W-T, Solin LJ, Vapiwala N. Incidence and patterns of distant metastases for patients with early-stage breast Cancer after breast conservation treatment. Clin Breast Cancer. 2013;13(2):88–94.PubMedCrossRef

53.

Chien AJ, Moasser MM. Cellular mechanisms of Resistance to anthracyclines and taxanes in Cancer: intrinsic and acquired. Semin Oncol. 2008;35:S1–14.PubMedCrossRef

54.

Popov C, Radic T, Haasters F, Prall W, Aszodi A, Gullberg D, et al. Integrins α2β1 and α11β1 regulate the survival of mesenchymal stem cells on collagen I. Cell Death Dis. 2011;2(7):e186–186.PubMedPubMedCentralCrossRef

55.

Bansal R, Nakagawa S, Yazdani S, Van Baarlen J, Venkatesh A, Koh AP, et al. Integrin alpha 11 in the regulation of the myofibroblast phenotype: implications for fibrotic diseases. Exp Mol Med. 2017;49(11):e396–396.PubMedPubMedCentralCrossRef

56.

Zeltz C, Alam J, Liu H, Erusappan PM, Hoschuetzky H, Molven A, et al. α11β1 integrin is induced in a subset of cancer-associated fibroblasts in desmoplastic tumor stroma and mediates in vitro cell migration. Cancers. 2019;11(6):765.PubMedPubMedCentralCrossRef

57.

Wu P, Wang Y, Wu Y, Jia Z, Song Y, Liang N. Expression and prognostic analyses of ITGA11, ITGB4 and ITGB8 in human non-small cell lung cancer. PeerJ. 2019;7:e8299.PubMedPubMedCentralCrossRef

58.

Pasini D, Bracken AP, Jensen MR, Denchi EL, Helin K. Suz12 is essential for mouse development and for EZH2 histone methyltransferase activity. EMBO J. 2004;23(20):4061–71.PubMedPubMedCentralCrossRef

59.

Konze KD, Ma A, Li F, Barsyte-Lovejoy D, Parton T, MacNevin CJ, et al. An orally bioavailable chemical probe of the lysine methyltransferases EZH2 and EZH1. ACS Chem Biol. 2013;8(6):1324–34.PubMedPubMedCentralCrossRef

60.

O’Meara MM, Simon JA. Inner workings and regulatory inputs that control polycomb repressive complex 2. Chromosoma. 2012;121:221–34.PubMedPubMedCentralCrossRef

61.

Ding X, Wang X, Sontag S, Qin J, Wanek P, Lin Q, et al. The polycomb protein Ezh2 impacts on induced pluripotent stem cell generation. Stem Cells Dev. 2014;23(9):931–40.PubMedCrossRef

62.

Tan J-z, Yan Y, Wang X-x, Jiang Y, Xu HE. EZH2: biology, disease, and structure-based drug discovery. Acta Pharmacol Sin. 2014;35(2):161–74.PubMedCrossRef

63.

Lund K, Adams P, Copland M. EZH2 in normal and malignant hematopoiesis. Leukemia. 2014;28(1):44–9.PubMedCrossRef

64.

Chaudhary P, Guragain D, Chang J-H, Kim J-A. TPH1 and 5-HT7 receptor overexpression leading to Gemcitabine-Resistance requires non-canonical permissive action of EZH2 in pancreatic ductal adenocarcinoma. Cancers. 2021;13(21):5305.PubMedPubMedCentralCrossRef

65.

Zhao Y, Wang X-X, Wu W, Long H, Huang J, Wang Z, et al. EZH2 regulates PD-L1 expression via HIF-1α in non-small cell lung cancer cells. Biochem Biophys Res Commun. 2019;517(2):201–9.PubMedCrossRef

66.

Li H, Wang H, Cui Y, Jiang W, Zhan H, Feng L, et al. EZH2 regulates pancreatic cancer cells through E2F1, GLI1, CDK3, and Mcm4. Hereditas. 2023;160(1):23.PubMedPubMedCentralCrossRef

67.

Katano M. Hedgehog signaling pathway as a therapeutic target in breast cancer. Cancer Lett. 2005;227(2):99–104.PubMedCrossRef

68.

Koga K, Nakamura M, Nakashima H, Akiyoshi T, Kubo M, Sato N, et al. Novel link between estrogen receptor α and hedgehog pathway in breast Cancer. Anticancer Res. 2008;28(2A):731–9.PubMed

69.

Im S, Choi HJ, Yoo C, Jung J-H, Jeon Y-W, Suh YJ, et al. Hedgehog related protein expression in breast Cancer: Gli-2 is Associated with poor overall survival. Korean J Pathol. 2013;47(2):116–23.PubMedPubMedCentralCrossRef

70.

Wang B, Yu T, Hu Y, Xiang M, Peng H, Lin Y et al. Prognostic role of Gli1 expression in breast cancer: a meta-analysis. Oncotarget 2017, 8(46).

71.

Glénisson M, Vacher S, Callens C, Susini A, Cizeron-Clairac G, Le Scodan R, et al. Identification of new candidate therapeutic target genes in triple-negative breast cancer. Genes Cancer. 2012;3(1):63–70.PubMedPubMedCentralCrossRef

72.

Shibue T, Weinberg RA. Integrin β1-focal adhesion kinase signaling directs the proliferation of metastatic cancer cells disseminated in the lungs. Proc Natl Acad Sci U S A. 2009;106(25):10290–5.PubMedPubMedCentralCrossRef

73.

Popova S, Lundgren-Åkerlund E, Wiig H, Gullberg D. Physiology and pathology of collagen receptors. Acta Physiol. 2007;190(3):179–87.CrossRef

74.

Erusappan P, Alam J, Lu N, Zeltz C, Gullberg D. Integrin α11 cytoplasmic tail is required for FAK activation to initiate 3D cell invasion and ERK-mediated cell proliferation. Sci Rep. 2019;9(1):15283.PubMedPubMedCentralCrossRef

Title: siRNA treatment targeting integrin α11 overexpressed via EZH2-driven axis inhibits drug-resistant breast cancer progression
Authors: Prakash Chaudhary
Kiran Yadav
Ho Jin Lee
Keon Wook Kang
Jongseo Mo
Jung-Ae Kim
Publication date: 01-12-2024
Publisher: BioMed Central
Keywords: Breast Cancer
Breast Cancer
Tamoxifen
Published in: Breast Cancer Research / Issue 1/2024
Electronic ISSN: 1465-542X
DOI: https://doi.org/10.1186/s13058-024-01827-4

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