Osteolytic effects of tumoral estrogen signaling in an estrogen receptor-positive breast cancer bone metastasis model

Abstract

Aim: Estrogen receptor α-positive (ER+) subtypes of breast cancer have the greatest predilection for forming osteolytic bone metastases (BMETs). Because tumor-derived factors mediate osteolysis, a possible role for tumoral ERα signaling in driving ER+ BMET osteolysis was queried using an estrogen (E₂)-dependent ER+ breast cancer BMET model.

Methods: Female athymic Foxn1^nu mice were inoculated with human ER+ MCF-7 breast cancer cells via the left cardiac ventricle post-E₂ pellet placement, and age- and dose-dependent E₂ effects on osteolytic ER+ BMET progression, as well as direct bone effects of E₂, were determined.

Results: Osteolytic BMETs, which did not form in the absence of E₂ supplementation, occurred with the same frequency in young (5-week-old) vs. skeletally mature (16-week-old) E₂ (0.72 mg)-treated mice, but were larger in young mice where anabolic bone effects of E₂ were greater. However, in mice of a single age and across a range of E₂ doses, anabolic E₂ bone effects were constant, while osteolytic ER+ BMET lesion incidence and size increased in an E₂ dose-dependent fashion. Osteoclasts in ER+ tumor-bearing (but not tumor-naive) mice increased in an E₂-dose dependent fashion at the bone-tumor interface, while histologic tumor size and proliferation did not vary with E₂ dose. E₂-inducible tumoral secretion of the osteolytic factor parathyroid hormone-related protein (PTHrP) was dose-dependent and mediated by ERα, with significantly greater levels of secretion from ER+ BMET-derived tumor cells.

Conclusion: These results suggest that tumoral ERα signaling may contribute to ER+ BMET-associated osteolysis, potentially explaining the greater predilection for ER+ tumors to form clinically-evident osteolytic BMETs.

Keywords

Breast cancer / estrogen receptor / bone metastasis / estradiol / osteolysis / osteoclasts / parathyroid hormone-related protein / bone

References

[1] Bray F,Soerjomataram I,Torre LA.Global cancer statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries.CA Cancer J Clin2018;68:394-424

[2] Macedo F,Pinho F.Bone metastases: an overview.Oncol Rev2017;11:321 PMCID:PMC5444408

[3] Kozlow W.Breast cancer metastasis to bone: mechanisms of osteolysis and implications for therapy.J Mammary Gland Biol Neoplasia2005;10:169-80

[4] Soni A,Hameed O.Breast cancer subtypes predispose the site of distant metastases.Am J Clin Pathol2015;143:471-8

[5] Hilton JF,Hopkins S.Acquisition of metastatic tissue from patients with bone metastases from breast cancer.Breast Cancer Res Treat2011;129:761-5

[6] Kamby C,Kristensen B.Oestrogen receptor status of primary breast carcinomas and their metastases. Relation to pattern of spread and survival after recurrence.Br J Cancer1989;60:252-7 PMCID:PMC2247047

[7] Aurilio G,Rizzo S.Discordant hormone receptor and human epidermal growth factor receptor 2 status in bone metastases compared to primary breast cancer.Acta Oncol2013;52:1649-56

[8] Guise TA.Molecular mechanisms of osteolytic bone metastases.Cancer2000;88:2892-8

[9] Coleman RE,Padhani AR.Bone metastases.Nat Rev Dis Primers2020;6:83

[10] Guise TA.The vicious cycle of bone metastases.J Musculoskelet Neuronal Interact2002;2:570-2

[11] Guise TA.Parathyroid hormone-related protein and bone metastases.Cancer1997;80:1572-80

[12] Reinert T.Optimal management of hormone receptor positive metastatic breast cancer in 2016.Ther Adv Med Oncol2015;7:304-20 PMCID:PMC4622303

[13] Breast Cancer Trialists' Collaborative Group (EBCTCG). Effects of chemotherapy and hormonal therapy for early breast cancer on recurrence and 15-year survival: an overview of the randomised trials.Lancet2005;365:1687-717

[14] Zhao H,Shangguan AJ.Aromatase expression and regulation in breast and endometrial cancer.J Mol Endocrinol2016;57:R19-33 PMCID:PMC5519084

[15] Sjögren K,Moverare-Skrtic S.Elevated aromatase expression in osteoblasts leads to increased bone mass without systemic adverse effects.J Bone Miner Res2009;24:1263-70

[16] Nilsson ME,Tivesten A.Measurement of a comprehensive sex steroid profile in rodent serum by high-sensitive gas chromatography-tandem mass spectrometry.Endocrinology2015;156:2492-502

[17] Osborne CK,Clark GM.Effect of estrogens and antiestrogens on growth of human breast cancer cells in athymic nude mice.Cancer Res1985;45:584-90

[18] Holen I,Nutter F.Oestrogen receptor positive breast cancer metastasis to bone: inhibition by targeting the bone microenvironment in vivo.Clin Exp Metastasis2016;33:211-24

[19] Ogba N,Bliesner BS.Luminal breast cancer metastases and tumor arousal from dormancy are promoted by direct actions of estradiol and progesterone on the malignant cells.Breast Cancer Res2014;16:48 PMCID:PMC4303198

[20] Sowder ME.Enrichment and detection of bone disseminated tumor cells in models of low tumor burden.Sci Rep2018;8:14299 PMCID:PMC6155169

[21] Canon J,Roudier M,Dougall WC.RANKL inhibition combined with tamoxifen treatment increases anti-tumor efficacy and prevents tumor-induced bone destruction in an estrogen receptor-positive breast cancer bone metastasis model.Breast Cancer Res Treat2012;135:771-80

[22] Fisher JL,Elliott J.Osteoprotegerin overexpression by breast cancer cells enhances orthotopic and osseous tumor growth and contrasts with that delivered therapeutically.Cancer Res2006;66:3620-8

[23] Gawrzak S,Gregorio S.MSK1 regulates luminal cell differentiation and metastatic dormancy in ER⁺ breast cancer.Nat Cell Biol2018;20:211-21

[24] Johnson RW,Olcina MM.Induction of LIFR confers a dormancy phenotype in breast cancer cells disseminated to the bone marrow.Nat Cell Biol2016;18:1078-89 PMCID:PMC5357601

[25] Pavlovic M,Rojo F.Enhanced MAF oncogene expression and breast cancer bone metastasis.J Natl Cancer Inst2015;107:djv256 PMCID:PMC4681582

[26] Thomas RJ,Yin JJ.Breast cancer cells interact with osteoblasts to support osteoclast formation.Endocrinology1999;140:4451-8

[27] Clements ME.PREX1 drives spontaneous bone dissemination of ER+ breast cancer cells.Oncogene2020;39:1318-34 PMCID:PMC7007387

[28] Ottewell PD,Brown HK.Zoledronic acid has differential antitumor activity in the pre- and postmenopausal bone microenvironment in vivo.Clin Cancer Res2014;20:2922-32 PMCID:PMC4040234

[29] Bord S,Beavan S.Estrogen receptors alpha and beta are differentially expressed in developing human bone.J Clin Endocrinol Metab2001;86:2309-14

[30] Braidman IP,Batra G,Saunders PT.Localization of estrogen receptor beta protein expression in adult human bone.J Bone Miner Res2001;16:214-20

[31] Rooney AM.Mouse models to evaluate the role of estrogen receptor α in skeletal maintenance and adaptation.Ann N Y Acad Sci2017;1410:85-92

[32] Manolagas SC,Almeida M.The role of estrogen and androgen receptors in bone health and disease.Nat Rev Endocrinol2013;9:699-712 PMCID:PMC3971652

[33] Khosla S.Regulation of bone metabolism by sex steroids.Cold Spring Harb Perspect Med2018;8:a031211 PMCID:PMC5749141

[34] Cheng JN,Whitman SA.Skeletal impact of 17β-estradiol in T cell-deficient mice: age-dependent bone effects and osteosarcoma formation.Clin Exp Metastasis2020;37:269-81 PMCID:PMC7731916

[35] Winding B,Høegh-andersen P,Tækker Foged N.Estradiol enhances osteolytic lesions in mice inoculated with human estrogen receptor-negative MDA-231 breast cancer cells in vivo.Breast Cancer Res Treat2003;78:205-16

[36] Cohen DJ,Verma A,Schwartz Z.Effect of 17β-estradiol on estrogen receptor negative breast cancer cells in an osteolytic mouse model.Steroids2019;142:28-33

[37] Ottewell PD,Brown HK.OPG-Fc inhibits ovariectomy-induced growth of disseminated breast cancer cells in bone.Int J Cancer2015;137:968-77

[38] Wang W,Yang X.Effects of letrozole on breast cancer micro-metastatic tumor growth in bone and lung in mice inoculated with murine 4T1 cells.Clin Exp Metastasis2016;33:475-85

[39] Price TT,Sivan A.Dormant breast cancer micrometastases reside in specific bone marrow niches that regulate their transit to and from bone.Sci Transl Med2016;8:340ra73

[40] Wang H,Gao X.The osteogenic niche promotes early-stage bone colonization of disseminated breast cancer cells.Cancer Cell2015;27:193-210 PMCID:PMC4326554

[41] Wright LE,Lukefahr AL.Curcuminoids block TGF-β signaling in human breast cancer cells and limit osteolysis in a murine model of breast cancer bone metastasis.J Nat Prod2013;76:316-21 PMCID:PMC3596492

[42] Guise TA,Taylor SD.Evidence for a causal role of parathyroid hormone-related protein in the pathogenesis of human breast cancer-mediated osteolysis.J Clin Invest1996;98:1544-9 PMCID:PMC507586

[43] Kunihiro AG,Frye JB,Schneider C.Curcumin, but not curcumin-glucuronide, inhibits Smad signaling in TGFβ-dependent bone metastatic breast cancer cells and is enriched in bone compared to other tissues.J Nutr Biochem2019;63:150-6 PMCID:PMC6296872

[44] Faul F,Lang AG.G*Power 3: a flexible statistical power analysis program for the social, behavioral, and biomedical sciences.Behav Res Methods2007;39:175-91

[45] Pearse G,Randall KJ.Urinary retention and cystitis associated with subcutaneous estradiol pellets in female nude mice.Toxicol Pathol2009;37:227-34

[46] Gakhar G,Andrews G,Nguyen TA.Hydronephrosis and urine retention in estrogen-implanted athymic nude mice.Vet Pathol2009;46:505-8

[47] Wright LE,Timmermann BN.Protection of trabecular bone in ovariectomized rats by turmeric (Curcuma longa L.) is dependent on extract composition.J Agric Food Chem2010;58:9498-504 PMCID:PMC2945868

[48] Funk JL,Wei H,Yocum DE.Synovium as a source of increased amino-terminal parathyroid hormone-related protein expression in rheumatoid arthritis. A possible role for locally produced parathyroid hormone-related protein in the pathogenesis of rheumatoid arthritis.J Clin Invest1998;101:1362-71 PMCID:PMC508713

[49] Dempster DW,Drezner MK.Standardized nomenclature, symbols, and units for bone histomorphometry: a 2012 update of the report of the ASBMR Histomorphometry Nomenclature Committee.J Bone Miner Res2013;28:2-17 PMCID:PMC3672237

[50] Haider MT,Dear TN,Brown HK.Modifying the osteoblastic niche with zoledronic acid in vivo-potential implications for breast cancer bone metastasis.Bone2014;66:240-50 PMCID:PMC4127787

[51] Streicher C,Andrukhova O.Estrogen regulates bone turnover by targeting RANKL expression in bone lining cells.Sci Rep2017;7:6460 PMCID:PMC5527119

[52] Florencio-Silva R,Sasso-Cerri E,Cerri PS.Biology of bone tissue: structure, function, and factors that influence bone cells.Biomed Res Int2015;2015:421746 PMCID:PMC4515490

[53] Brown HK,Evans CA.Location matters: osteoblast and osteoclast distribution is modified by the presence and proximity to breast cancer cells in vivo.Clin Exp Metastasis2012;29:927-38

[54] Meng X,Lee P.Myeloid-specific TGF-β signaling in bone promotes basic-FGF and breast cancer bone metastasis.Oncogene2016;35:2370-8

[55] Biswas S,Alvarez J.Anti-transforming growth factor ß antibody treatment rescues bone loss and prevents breast cancer metastasis to bone.PLoS One2011;6:e27090 PMCID:PMC3214031

[56] Haisenleder DJ,Marcinko ES,Marshall JC.Estimation of estradiol in mouse serum samples: evaluation of commercial estradiol immunoassays.Endocrinology2011;152:4443-7 PMCID:PMC3198998

[57] Bouxsein ML,Shultz KL,Rosen CJ.Ovariectomy-induced bone loss varies among inbred strains of mice.J Bone Miner Res2005;20:1085-92

[58] Gérard C,Tskitishvili E.Combined estrogenic and anti-estrogenic properties of estetrol on breast cancer may provide a safe therapeutic window for the treatment of menopausal symptoms.Oncotarget2015;6:17621-36 PMCID:PMC4627333

[59] Wright LE,Rucci N.Murine models of breast cancer bone metastasis.Bonekey Rep2016;5:804 PMCID:PMC5108088

[60] Khosla S,Monroe DG.Estrogen and the skeleton.Trends Endocrinol Metab2012;23:576-81 PMCID:PMC3424385

[61] Fantozzi A.Mouse models of breast cancer metastasis.Breast Cancer Res2006;8:212. PMCID:PMC1779475

[62] Powell GJ,Danks JA.Localization of parathyroid hormone-related protein in breast cancer metastases: increased incidence in bone compared with other sites.Cancer Res1991;51:3059-61

[63] Southby J,Danks JA.Immunohistochemical localization of parathyroid hormone-related protein in human breast cancer.Cancer Res1990;50:7710-6

[64] Sun J,Harrington WR,Katzenellenbogen JA.Antagonists selective for estrogen receptor alpha.Endocrinology2002;143:941-7

[65] Kraichely DM,Katzenellenbogen JA.Conformational changes and coactivator recruitment by novel ligands for estrogen receptor-alpha and estrogen receptor-beta: correlations with biological character and distinct differences among SRC coactivator family members.Endocrinology2000;141:3534-45

[66] Li S,Weinhandl ED.Estimated number of prevalent cases of metastatic bone disease in the US adult population.Clin Epidemiol2012;4:87-93 PMCID:PMC3345874

[67] Mundy GR.Metastasis: metastasis to bone: causes, consequences and therapeutic opportunities.Nat Rev Cancer2002;2:584-93.

[68] Svendsen H,Sværke C.Hospital visits among women with skeletal-related events secondary to breast cancer and bone metastases: a nationwide population-based cohort study in Denmark.Clin Epidemiol2013;5:97. PMCID:PMC3616605

[69] Haque R,Inzhakova G.Impact of breast cancer subtypes and treatment on survival: an analysis spanning two decades.Cancer Epidemiol Biomarkers Prev2012;21:1848-55. PMCID:PMC3467337

[70] Colzani E,Liljegren A.Time-dependent risk of developing distant metastasis in breast cancer patients according to treatment, age and tumour characteristics.Br J Cancer2014;110:1378-84. PMCID:PMC3950882

[71] Turner NC,Loibl S.Advances in the treatment of advanced oestrogen-receptor-positive breast cancer.Lancet2017;389:2403-14.

[72] Portman N,Carson E,Lim E.Overcoming CDK4/6 inhibitor resistance in ER-positive breast cancer.Endocr Relat Cancer2019;26:R15-30.

[73] Yoneda T,Hiraga T,Nishimura R.A bone-seeking clone exhibits different biological properties from the MDA-MB-231 parental human breast cancer cells and a brain-seeking clone in vivo and in vitro.J Bone Miner Res2001;16:1486-95.

[74] Funk JL.Regulation of parathyroid hormone-related protein expression in MCF-7 breast carcinoma cells by estrogen and antiestrogens.Biochem Biophys Res Commun1998;251:849-54.

[75] Black DM.Clinical practice: postmenopausal osteoporosis.N Engl J Med2016;374:254-62

[76] Cancer Stat Facts: Female Breast Cancer Subtypes. Natl Cancer Inst Surveillance, Epidemiol End Results Progr n.d. https://seer.cancer.gov/statfacts/html/breast-subtypes.html. [Last accessed on 30 Mar 2021]

[77] Braun S,Naume B.A pooled analysis of bone marrow micrometastasis in breast cancer.N Engl J Med2005;353:793-802.

[78] Braun S,Marth C.The prognostic impact of bone marrow micrometastases in women with breast cancer.Cancer Invest2009;27:598-603.

[79] Hussein O.Breast cancer at bone metastatic sites: recent discoveries and treatment targets.J Cell Commun Signal2011;5:85-99. PMCID:PMC3088795

[80] Falck AK,Ingvar C.Analysis of and prognostic information from disseminated tumour cells in bone marrow in primary breast cancer: a prospective observational study.BMC Cancer2012;12:403 PMCID:PMC3488538

[81] Brown HK,Ottewell PD.Parathyroid hormone (PTH) increases skeletal tumour growth and alters tumour distribution in an in vivo model of breast cancer.Int J Mol Sci2018;19:2920. PMCID:PMC6213905

[82] Swami S,Bettinson LA.Prevention of breast cancer skeletal metastases with parathyroid hormone.JCI Insight2017;2:90874 PMCID:PMC5621896

[83] Jiang G,Yazdanparast A.Comprehensive comparison of molecular portraits between cell lines and tumors in breast cancer.BMC Genomics2016;17:525 PMCID:PMC5001206

[84] Sasaki A,Story B.Bisphosphonate risedronate reduces metastatic human breast cancer burden in bone in nude mice.Cancer Res1995;55:3551-7

[85] Holen I,Morrissey B.In vivo models in breast cancer research: progress, challenges and future directions.Dis Model Mech2017;10:359-71. PMCID:PMC5399571

[86] Murayama T.Patient-derived Xenograft models of breast cancer and their application.Cells2019;8:621 PMCID:PMC6628218

[87] Dobrolecki LE,Alferez DG.Patient-derived xenograft (PDX) models in basic and translational breast cancer research.Cancer Metastasis Rev2016;35:547-73 PMCID:PMC5396460

[88] Matthews SB.Steroid hormone receptor positive breast cancer patient-derived Xenografts.Horm Cancer2017;8:4-15 PMCID:PMC5291783

[89] Yin JJ,Käkönen SM.A causal role for endothelin-1 in the pathogenesis of osteoblastic bone metastases.Proc Natl Acad Sci U S A2003;100:10954-9 PMCID:PMC196909