Abstract
It is becoming increasingly evident that progression and metastasis of solid cancers is driven by the interaction of oncogene-transformed cancer cells and non-malignant host cells in the tumor stroma. In this process, the immune system contributes a complex set of highly important pro- and antitumor effects, which are not readily recapitulated by commonly used xenograft cancer models in immunodeficient mice.
Therefore, we provide protocols for isolation of primary tumor cells from the MMTV-PymT mouse model for metastasizing breast cancer and their resubmission to congenic immunocompetent mice by orthotopic transplantation into the mammary gland or different routes of injection to induce organ-specific experimental metastasis, including intravenous, intracardiac, and caudal artery injection of tumor cells. Moreover, we describe protocols for sensitive detection and quantification of the metastatic burden.
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References
Puente XS, Sanchez LM, Overall CM, Lopez-Otin C (2003) Human and mouse proteases: a comparative genomic approach. Nat Rev Genet 4:544–558
Turk V, Stoka V, Vasiljeva O et al (2012) Cysteine cathepsins: from structure, function and regulation to new frontiers. Biochim Biophys Acta 1824:68–88
Reinheckel T, Peters C, Krüger A et al (2012) Differential impact of cysteine Cathepsins on genetic mouse models of De novo carcinogenesis: Cathepsin B as emerging therapeutic target. Front Pharmacol 3
Vidak E, Javoršek U, Vizovišek M, Turk B (2019) Cysteine Cathepsins and their extracellular roles: shaping the microenvironment. Cell 8:264
Bengsch F, Buck A, Günther SC et al (2013) Cell type-dependent pathogenic functions of overexpressed human cathepsin B in murine breast cancer progression. Oncogene 33:4474–4484. https://doi.org/10.1038/onc.2013.395
Dennemärker J, Lohmüller T, Mayerle J et al (2010) Deficiency for the cysteine protease cathepsin L promotes tumor progression in mouse epidermis. Oncogene 29:1611–1621
Sevenich L, Schurigt U, Sachse K et al (2010) Synergistic antitumor effects of combined cathepsin B and cathepsin Z deficiencies on breast cancer progression and metastasis in mice. Proc Natl Acad Sci U S A 107:2497–2502
Vasiljeva O, Papazoglou A, Krüger A et al (2006) Tumor cell-derived and macrophage-derived cathepsin B promotes progression and lung metastasis of mammary cancer. Cancer Res 66:5242–5250
Gocheva V, Zeng W, Ke D et al (2006) Distinct roles for cysteine cathepsin genes in multistage tumorigenesis. Genes Dev 20:543–556
Vasiljeva O, Korovin M, Gajda M et al (2008) Reduced tumour cell proliferation and delayed development of high-grade mammary carcinomas in cathepsin B-deficient mice. Oncogene 27:4191–4199
Sevenich L, Werner F, Gajda M et al (2011) Transgenic expression of human cathepsin B promotes progression and metastasis of polyoma-middle-T-induced breast cancer in mice. Oncogene 30:54–64
Sevenich L, Bowman RL, Mason SD et al (2014) Analysis of tumour- and stroma-supplied proteolytic networks reveals a brain-metastasis-promoting role for cathepsin S. Nat Cell Biol 16:876–888
Tholen M, Wolanski J, Stolze B et al (2015) Stress-resistant translation of Cathepsin L mRNA in breast cancer progression. J Biol Chem 290:15758–15769
Ziegler PK, Bollrath J, Pallangyo CK et al (2018) Mitophagy in intestinal epithelial cells triggers adaptive immunity during tumorigenesis. Cell 174:88–101
Guy CT, Cardiff RD, Muller WJ (1992) Induction of mammary tumors by expression of polyomavirus middle T oncogene: a transgenic mouse model for metastatic disease. Mol Cell Biol 12:954–961
Elston CW, Ellis IO (2002) Pathological prognostic factors in breast cancer. I. the value of histological grade in breast cancer: experience from a large study with long-term follow-up. Histopathology 41:154–161
Almholt K, Lund LR, Rygaard J et al (2005) Reduced metastasis of transgenic mammary cancer in urokinase-deficient mice. Int J Cancer 113:525–532
Hüsemann Y, Geigl JB, Schubert F et al (2008) Systemic spread is an early step in breast cancer. Cancer Cell 13:58–68
Kang Y, Siegel PM, Shu W et al (2003) A multigenic program mediating breast cancer metastasis to bone. Cancer Cell 3:537–549
Bos PD, Zhang XH-F, Nadal C et al (2009) Genes that mediate breast cancer metastasis to the brain. Nature 459:1005–1009
Kuchimaru T, Kataoka N, Nakagawa K et al (2018) A reliable murine model of bone metastasis by injecting cancer cells through caudal arteries. Nat Commun 9:2981
Bowman RL, Klemm F, Akkari L et al (2016) Macrophage ontogeny underlies differences in tumor-specific education in brain malignancies. Cell Rep 17:2445–2459
Chae WH, Niesel K, Schulz M et al (2019) Evaluating magnetic resonance spectroscopy as a tool for monitoring therapeutic response of whole brain radiotherapy in a mouse model for breast-to-brain metastasis. Front Oncol 9:1324
Lin EY, Nguyen V, Russell RG, Pollard JW (2001) Colony-stimulating factor 1 promotes progression of mammary tumors to malignancy. J Exp Med 193:727–740
Shree T, Olson OC, Elie BT et al (2011) Macrophages and cathepsin proteases blunt chemotherapeutic response in breast cancer. Genes Dev 25:2465–2479
Qian B-Z, Li J, Zhang H et al (2011) CCL2 recruits inflammatory monocytes to facilitate breast-tumour metastasis. Nature 475:222–225
Guerin MV, Finisguerra V, Van den Eynde BJ et al (2020) Preclinical murine tumor models: a structural and functional perspective. elife 9:e50740
Walsh NC, Kenney LL, Jangalwe S et al (2017) Humanized mouse models of clinical disease. Annu Rev Pathol Mech Dis 12:187–215
Tu W, Zheng J (2016) Application of humanized mice in immunological research. Methods Mol Biol 1371:157–176
Hillebrand LE, Wickberg SM, Gomez-Auli A et al (2019) MMP14 empowers tumor-initiating breast cancer cells under hypoxic nutrient-depleted conditions. FASEB J 33:4124–4140
Acknowledgment
Grant Support: TR is supported by the Deutsche Forschungsgemeinschaft (DFG) SFB 850 subproject B7, GRK 2606 “ProtPath” and the German Cancer Consortium (DKTK) programs Exploitation of Oncogenic Mechanisms project L627 and Radiation Oncology and Imaging project “eRadioImage”. LS is supported by the Deutsche Krebshilfe Max Eder Junior Group Leader Program (70111752) and the Deutsche Forschungsgemeinschaft SPP2084 μbone.
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Vasiljeva, O., Sevenich, L., Reinheckel, T. (2021). Analyzing the Role of Proteases in Breast Cancer Progression and Metastasis Using Primary Cells from Transgenic Oncomice. In: Stein, U.S. (eds) Metastasis. Methods in Molecular Biology, vol 2294. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1350-4_20
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DOI: https://doi.org/10.1007/978-1-0716-1350-4_20
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