Α Humanized RANKL Transgenic Mouse Model of Progestin-Induced Mammary Carcinogenesis for Evaluation of Novel Therapeutics
Abstract
:Simple Summary
Abstract
1. Introduction
2. Materials and Methods
2.1. Mouse Husbandry
2.2. MPA/DMBA-Induced Mammary Carcinogenesis
2.3. Administration of OPG-Fc and Denosumab
2.4. In Vivo Imaging
2.5. Whole-Mount Analysis, Carmine-Alum Staining of Mammary Glands
2.6. Histology and Immunohistochemistry
2.7. MicroCT Tomography of Mammary Glands
2.8. MicroCT Tomography Analysis of Femurs
2.9. RNA Isolation and qPCR Analysis
2.10. Multiplex ELISA
2.11. Statistical Analysis
3. Results
3.1. Comparison of MPA/DMBA-Driven Mammary Carcinogenesis between Wild-Type Mice and Transgenic Mice Overexpressing Human RANKL
3.2. Treatment with OPG-Fc Prevents MPA/DMBA-Induced Mammary Carcinogenesis
3.3. Generation of a Humanized RANKL Transgenic Mouse Model of Mammary Carcinogenesis
3.4. Prophylactic Denosumab Treatment Attenuates the Development of MPA/DMBA-Induced Mammary Carcinogenesis in HumTgRANKL Mice
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
- Sigl, V.; Jones, L.P.; Penninger, J.M. RANKL/RANK: From Bone Loss to the Prevention of Breast Cancer. Open Biol. 2016, 6, 160230. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Henderson, B.E.; Ross, R.K.; Judd, H.L.; Krailo, M.D.; Pike, M.C. Do Regular Ovulatory Cycles Increase Breast Cancer Risk? Cancer 1985, 56, 1206–1208. [Google Scholar] [CrossRef] [PubMed]
- Söderqvist, G. Effects of Sex Steroids on Proliferation in Normal Mammary Tissue. Ann. Med. 1998, 30, 511–524. [Google Scholar] [CrossRef] [PubMed]
- Banks, E.; Beral, V.; Bull, D.; Reeves, G.; Austoker, J.; English, R.; Patnick, J.; Peto, R.; Vessey, M.; Wallis, M.; et al. Breast Cancer and Hormone-Replacement Therapy in the Million Women Study. Lancet 2003, 362, 419–427. [Google Scholar] [CrossRef]
- Beverly, R.; Volkar, J. Risks and Benefits of Estrogen Plus Progestin in Healthy Postmenopausal Women. In 50 Studies Every Obstetrician-Gynecologist Should Know; Oxford Academic: Oxford, UK, 2021; Volume 288, pp. 262–266. Available online: https://academic.oup.com/book/32722/chapter-abstract/272469459?redirectedFrom=fulltext (accessed on 2 August 2023).
- Olver, I.N. Prevention of Breast Cancer. Med. J. Aust. 2016, 205, 475–479. [Google Scholar] [CrossRef]
- Narod, S.A. Hormone Replacement Therapy and the Risk of Breast Cancer. Nat. Rev. Clin. Oncol. 2011, 8, 669–676. [Google Scholar] [CrossRef]
- Fuller, K.; Wong, B.; Fox, S.; Choi, Y.; Chambers, T.J. TRANCE Is Necessary and Sufficient for Osteoblast-Mediated Activation of Bone Resorption in Osteoclasts. J. Exp. Med. 1998, 188, 997–1001. [Google Scholar] [CrossRef] [PubMed]
- Takahashi, N.; Udagawa, N.; Suda, T. A New Member of Tumor Necrosis Factor Ligand Family, ODF/OPGL/TRANCE/RANKL, Regulates Osteoclast Differentiation and Function. Biochem. Biophys. Res. Commun. 1999, 256, 449–455. [Google Scholar] [CrossRef]
- Anandarajah, A.P. Role of RANKL in Bone Diseases. Trends Endocrinol. Metab. 2009, 20, 88–94. [Google Scholar] [CrossRef]
- Silva-Fernández, L.; Rosario, M.P.; Martínez-López, J.A.; Carmona, L.; Loza, E. Denosumab for the Treatment of Osteoporosis: A Systematic Literature Review. Reumatol. Clin. 2013, 9, 42–52. [Google Scholar] [CrossRef] [PubMed]
- Joshi, P.A.; Jackson, H.W.; Beristain, A.G.; Di Grappa, M.A.; Mote, P.A.; Clarke, C.L.; Stingl, J.; Waterhouse, P.D.; Khokha, R. Progesterone Induces Adult Mammary Stem Cell Expansion. Nature 2010, 465, 803–807. [Google Scholar] [CrossRef]
- Asselin-Labat, M.-L.; Vaillant, F.; Sheridan, J.M.; Pal, B.; Wu, D.; Simpson, E.R.; Yasuda, H.; Smyth, G.K.; Martin, T.J.; Lindeman, G.J.; et al. Control of Mammary Stem Cell Function by Steroid Hormone Signalling. Nature 2010, 465, 798–802. [Google Scholar] [CrossRef] [PubMed]
- Fernandez-Valdivia, R.; Lydon, J.P. From the Ranks of Mammary Progesterone Mediators, RANKL Takes the Spotlight. Mol. Cell. Endocrinol. 2012, 357, 91–100. [Google Scholar] [CrossRef] [Green Version]
- Fata, J.E.; Kong, Y.Y.; Li, J.; Sasaki, T.; Irie-Sasaki, J.; Moorehead, R.A.; Elliott, R.; Scully, S.; Voura, E.B.; Lacey, D.L.; et al. The Osteoclast Differentiation Factor Osteoprotegerin-Ligand Is Essential for Mammary Gland Development. Cell 2000, 103, 41–50. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Van Poznak, C.; Cross, S.S.; Saggese, M.; Hudis, C.; Panageas, K.S.; Norton, L.; Coleman, R.E.; Holen, I. Expression of Osteoprotegerin (OPG), TNF Related Apoptosis Inducing Ligand (TRAIL), and Receptor Activator of Nuclear Factor ΚB Ligand (RANKL) in Human Breast Tumours. J. Clin. Pathol. 2006, 59, 56–63. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Schramek, D.; Leibbrandt, A.; Sigl, V.; Kenner, L.; Pospisilik, J.A.; Lee, H.J.; Hanada, R.; Joshi, P.A.; Aliprantis, A.; Glimcher, L.; et al. Osteoclast Differentiation Factor RANKL Controls Development of Progestin-Driven Mammary Cancer. Nature 2010, 468, 98–102. [Google Scholar] [CrossRef] [Green Version]
- Infante, M.; Fabi, A.; Cognetti, F.; Gorini, S.; Caprio, M.; Fabbri, A. RANKL/RANK/OPG System beyond Bone Remodeling: Involvement in Breast Cancer and Clinical Perspectives. J. Exp. Clin. Cancer Res. 2019, 38, 12. [Google Scholar] [CrossRef] [Green Version]
- Gonzalez-Suarez, E.; Jacob, A.P.; Jones, J.; Miller, R.; Roudier-Meyer, M.P.; Erwert, R.; Pinkas, J.; Branstetter, D.; Dougall, W.C. RANK Ligand Mediates Progestin-Induced Mammary Epithelial Proliferation and Carcinogenesis. Nature 2010, 468, 103–107. [Google Scholar] [CrossRef]
- Rinotas, V.; Niti, A.; Dacquin, R.; Bonnet, N.; Stolina, M.; Han, C.-Y.; Kostenuik, P.; Jurdic, P.; Ferrari, S.; Douni, E. Novel Genetic Models of Osteoporosis by Overexpression of Human RANKL in Transgenic Mice. J. Bone Miner. Res. 2014, 29, 1158–1169. [Google Scholar] [CrossRef]
- Douni, E.; Rinotas, V.; Makrinou, E.; Zwerina, J.; Penninger, J.M.; Eliopoulos, E.; Schett, G.; Kollias, G. A RANKL G278R Mutation Causing Osteopetrosis Identifies a Functional Amino Acid Essential for Trimer Assembly in RANKL and TNF. Hum. Mol. Genet. 2012, 21, 784–798. [Google Scholar] [CrossRef] [Green Version]
- Kelloff, G.J.; Hoffman, J.M.; Johnson, B.; Scher, H.I.; Siegel, B.A.; Cheng, E.Y.; Cheson, B.D.; O’Shaughnessy, J.; Guyton, K.Z.; Mankoff, D.A.; et al. Progress and Promise of FDG-PET Imaging for Cancer Patient Management and Oncologic Drug Development. Clin. Cancer Res. 2005, 11, 2785–2808. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Nanni, C.; Pettinato, C.; Ambrosini, V.; Spinelli, A.; Trespidi, S.; Rubello, D.; Al-Nahhas, A.; Franchi, R.; Alavi, A.; Fanti, S. Retro-Orbital Injection Is an Effective Route for Radiopharmaceutical Administration in Mice during Small-Animal PET Studies. Nucl. Med. Commun. 2007, 28, 547–553. [Google Scholar] [CrossRef] [PubMed]
- Kolokotroni, A.; Gkikopoulou, E.; Rinotas, V.; Douni, E. Phosphotungstic Acid-Enhanced Microcomputed Tomography for Quantitative Visualization of Mouse Mammary Gland Morphology. J. Med. Imaging 2023, 10, 1–16. [Google Scholar] [CrossRef]
- Bouxsein, M.L.; Boyd, S.K.; Christiansen, B.A.; Guldberg, R.E.; Jepsen, K.J.; Müller, R. Guidelines for Assessment of Bone Microstructure in Rodents Using Micro-Computed Tomography. J. Bone Miner. Res. 2010, 25, 1468–1486. [Google Scholar] [CrossRef]
- Li, G.-W.; Chang, S.-X.; Fan, J.-Z.; Tian, Y.-N.; Xu, Z.; He, Y.-M. Marrow Adiposity Recovery after Early Zoledronic Acid Treatment of Glucocorticoid-Induced Bone Loss in Rabbits Assessed by Magnetic Resonance Spectroscopy. Bone 2013, 52, 668–675. [Google Scholar] [CrossRef]
- Poussin, C.; Mathis, C.; Alexopoulos, L.G.; Messinis, D.E.; Dulize, R.H.J.; Belcastro, V.; Melas, I.N.; Sakellaropoulos, T.; Rhrissorrakrai, K.; Bilal, E.; et al. The Species Translation Challenge—A Systems Biology Perspective on Human and Rat Bronchial Epithelial Cells. Sci. Data 2014, 1, 140009. [Google Scholar] [CrossRef] [Green Version]
- Hu, H.; Wang, J.; Gupta, A.; Shidfar, A.; Branstetter, D.; Lee, O.; Ivancic, D.; Sullivan, M.; Chatterton, R.T.; Dougall, W.C.; et al. RANKL Expression in Normal and Malignant Breast Tissue Responds to Progesterone and Is Up-Regulated during the Luteal Phase. Breast Cancer Res. Treat. 2014, 146, 515–523. [Google Scholar] [CrossRef] [PubMed]
- Cardiff, R.D.; Anver, M.R.; Gusterson, B.A.; Hennighausen, L.; Jensen, R.A.; Merino, M.J.; Rehm, S.; Russo, J.; Tavassoli, F.A.; Wakefield, L.M.; et al. The Mammary Pathology of Genetically Engineered Mice: The Consensus Report and Recommendations from the Annapolis Meeting. Oncogene 2000, 19, 968–988. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Miller, R.E.; Branstetter, D.; Armstrong, A.; Kennedy, B.; Jones, J.; Cowan, L.; Bussiere, J.; Dougall, W.C. Receptor Activator of NF-ΚB Ligand Inhibition Suppresses Bone Resorption and Hypercalcemia but Does Not Affect Host Immune Responses to Influenza Infection. J. Immunol. 2007, 179, 266–274. [Google Scholar] [CrossRef] [Green Version]
- Ominsky, M.S.; Kostenuik, P.J.; Cranmer, P.; Smith, S.Y.; Atkinson, J.E. The RANKL Inhibitor OPG-Fc Increases Cortical and Trabecular Bone Mass in Young Gonad-Intact Cynomolgus Monkeys. Osteoporos. Int. 2007, 18, 1073–1082. [Google Scholar] [CrossRef]
- Ominsky, M.S.; Li, X.; Asuncion, F.J.; Barrero, M.; Warmington, K.S.; Dwyer, D.; Stolina, M.; Geng, Z.; Grisanti, M.; Tan, H.L.; et al. RANKL Inhibition with Osteoprotegerin Increases Bone Strength by Improving Cortical and Trabecular Bone Architecture in Ovariectomized Rats. J. Bone Miner. Res. 2008, 23, 672–682. [Google Scholar] [CrossRef] [PubMed]
- Yue, Z.; Yuan, Z.; Zeng, L.; Wang, Y.; Lai, L.; Li, J.; Sun, P.; Xue, X.; Qi, J.; Yang, Z.; et al. LGR4 Modulates Breast Cancer Initiation, Metastasis, and Cancer Stem Cells. FASEB J. 2018, 32, 2422–2437. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Giannakeas, V.; Cadarette, S.M.; Ban, J.K.; Lipscombe, L.; Narod, S.A.; Kotsopoulos, J. Denosumab and Breast Cancer Risk in Postmenopausal Women: A Population-Based Cohort Study. Br. J. Cancer 2018, 119, 1421–1427. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Sigl, V.; Owusu-Boaitey, K.; Joshi, P.A.; Kavirayani, A.; Wirnsberger, G.; Novatchkova, M.; Kozieradzki, I.; Schramek, D.; Edokobi, N.; Hersl, J.; et al. RANKL/RANK Control Brca1 Mutation-Driven Mammary Tumors. Cell Res. 2016, 26, 761–774. [Google Scholar] [CrossRef] [Green Version]
- Nolan, E.; Vaillant, F.; Branstetter, D.; Pal, B.; Giner, G.; Whitehead, L.; Lok, S.W.; Mann, G.B.; Rohrbach, K.; Huang, L.Y.; et al. RANK Ligand as a Potential Target for Breast Cancer Prevention in BRCA1-Mutation Carriers. Nat. Med. 2016, 22, 933–939. [Google Scholar] [CrossRef]
- Singer, C.F. Nonsurgical Prevention Strategies in BRCA1 and BRCA2 Mutation Carriers. Breast Care 2021, 16, 144–148. [Google Scholar] [CrossRef]
- Gnant, M.; Pfeiler, G.; Steger, G.G.; Egle, D.; Greil, R.; Fitzal, F.; Wette, V.; Balic, M.; Haslbauer, F.; Melbinger-Zeinitzer, E.; et al. Adjuvant Denosumab in Postmenopausal Patients with Hormone Receptor-Positive Breast Cancer (ABCSG-18): Disease-Free Survival Results from a Randomised, Double-Blind, Placebo-Controlled, Phase 3 Trial. Lancet Oncol. 2019, 20, 339–351. [Google Scholar] [CrossRef]
- Coleman, R.; Finkelstein, D.M.; Barrios, C.; Martin, M.; Iwata, H.; Hegg, R.; Glaspy, J.; Periañez, A.M.; Tonkin, K.; Deleu, I.; et al. Adjuvant Denosumab in Early Breast Cancer (D-CARE): An International, Multicentre, Randomised, Controlled, Phase 3 Trial. Lancet Oncol. 2020, 21, 60–72. [Google Scholar] [CrossRef]
- Coleman, R.; Zhou, Y.; Jandial, D.; Cadieux, B.; Chan, A. Bone Health Outcomes from the International, Multicenter, Randomized, Phase 3, Placebo-Controlled D-CARE Study Assessing Adjuvant Denosumab in Early Breast Cancer. Adv. Ther. 2021, 38, 4569–4580. [Google Scholar] [CrossRef]
- Deligiorgi, M.V.; Panayiotidis, M.I.; Trafalis, D.T. Repurposing Denosumab in Breast Cancer beyond Prevention of Skeletal Related Events: Could Nonclinical Data Be Translated into Clinical Practice? Expert Rev. Clin. Pharmacol. 2020, 13, 1235–1252. [Google Scholar] [CrossRef]
- Wu, X.; Li, F.; Dang, L.; Liang, C.; Lu, A.; Zhang, G. RANKL/RANK System-Based Mechanism for Breast Cancer Bone Metastasis and Related Therapeutic Strategies. Front. Cell Dev. Biol. 2020, 8, 76. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Cao, Y.; Bonizzi, G.; Seagroves, T.N.; Greten, F.R.; Johnson, R.; Schmidt, E.V.; Karin, M. IKKα Provides an Essential Link between RANK Signaling and Cyclin D1 Expression during Mammary Gland Development. Cell 2001, 107, 763–775. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Ciscar, M.; Trinidad, E.M.; Perez-Montoyo, H.; Alsaleem, M.; Jimenez-Santos, M.J.; Toss, M.; Sanz-Moreno, A.; Vethencourt, A.; Perez-Chacon, G.; Petit, A.; et al. RANK Is an Independent Biomarker of Poor Prognosis in Estrogen Receptor-Negative Breast Cancer and a Therapeutic Target in Patient-Derived Xenografts. EMBO Mol. Med. 2023, 15, e16715. [Google Scholar] [CrossRef] [PubMed]
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Kolokotroni, A.; Gkikopoulou, E.; Rinotas, V.; Ntari, L.; Zareifi, D.; Rouchota, M.; Sarpaki, S.; Lymperopoulos, I.; Alexopoulos, L.G.; Loudos, G.; et al. Α Humanized RANKL Transgenic Mouse Model of Progestin-Induced Mammary Carcinogenesis for Evaluation of Novel Therapeutics. Cancers 2023, 15, 4006. https://doi.org/10.3390/cancers15154006
Kolokotroni A, Gkikopoulou E, Rinotas V, Ntari L, Zareifi D, Rouchota M, Sarpaki S, Lymperopoulos I, Alexopoulos LG, Loudos G, et al. Α Humanized RANKL Transgenic Mouse Model of Progestin-Induced Mammary Carcinogenesis for Evaluation of Novel Therapeutics. Cancers. 2023; 15(15):4006. https://doi.org/10.3390/cancers15154006
Chicago/Turabian StyleKolokotroni, Anthi, Evi Gkikopoulou, Vagelis Rinotas, Lydia Ntari, Danae Zareifi, Maritina Rouchota, Sophia Sarpaki, Ilias Lymperopoulos, Leonidas G. Alexopoulos, George Loudos, and et al. 2023. "Α Humanized RANKL Transgenic Mouse Model of Progestin-Induced Mammary Carcinogenesis for Evaluation of Novel Therapeutics" Cancers 15, no. 15: 4006. https://doi.org/10.3390/cancers15154006