Tumorigenic WAP-T Mouse Mammary Carcinoma Cells: A Model for a Self-Reproducing Homeostatic Cancer Cell System

Wegwitz, Florian; Kluth, Mark-Andreas; Mänz, Claudia; Otto, Benjamin; Gruner, Katharina; Heinlein, Christina; Kühl, Marion; Warnecke, Gabriele; Schumacher, Udo; Deppert, Wolfgang; Tolstonog, Genrich V.

doi:10.1371/journal.pone.0012103

Cited by 25 publications

(64 citation statements)

References 77 publications

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“…Despite the recently described cooperation between Slug and Sox 9 in mammary gland [29], we found that cell territories expressing these factors were very distinct in mammary tubules. Sox10 upregulation in SlugKO mice is noticeable considering the basal-like distribution previously reported [35] and the putative role in mammary cell fating [36].…”

Section: Discussionsupporting

confidence: 53%

Slug Controls Stem/Progenitor Cell Growth Dynamics during Mammary Gland Morphogenesis

et al. 2012

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BackgroundMorphogenesis results from the coordination of distinct cell signaling pathways controlling migration, differentiation, apoptosis, and proliferation, along stem/progenitor cell dynamics. To decipher this puzzle, we focused on epithelial-mesenchymal transition (EMT) “master genes”. EMT has emerged as a unifying concept, involving cell-cell adhesion, migration and apoptotic pathways. EMT also appears to mingle with stemness. However, very little is known on the physiological role and relevance of EMT master-genes. We addressed this question during mammary morphogenesis. Recently, a link between Slug/Snai2 and stemness has been described in mammary epithelial cells, but EMT master genes actual localization, role and targets during mammary gland morphogenesis are not known and we focused on this basic question.Methodology/Principal FindingsUsing a Slug–lacZ transgenic model and immunolocalization, we located Slug in a distinct subpopulation covering about 10–20% basal cap and duct cells, mostly cycling cells, coexpressed with basal markers P-cadherin, CK5 and CD49f. During puberty, Slug-deficient mammary epithelium exhibited a delayed development after transplantation, contained less cycling cells, and overexpressed CK8/18, ER, GATA3 and BMI1 genes, linked to luminal lineage. Other EMT master genes were overexpressed, suggesting compensation mechanisms. Gain/loss-of-function in vitro experiments confirmed Slug control of mammary epithelial cell luminal differentiation and proliferation. In addition, they showed that Slug enhances specifically clonal mammosphere emergence and growth, cell motility, and represses apoptosis. Strikingly, Slug-deprived mammary epithelial cells lost their potential to generate secondary clonal mammospheres.Conclusions/SignificanceWe conclude that Slug pathway controls the growth dynamics of a subpopulation of cycling progenitor basal cells during mammary morphogenesis. Overall, our data better define a key mechanism coordinating cell lineage dynamics and morphogenesis, and provide physiological relevance to broadening EMT pathways.

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Section: Discussionsupporting

confidence: 53%

Slug Controls Stem/Progenitor Cell Growth Dynamics during Mammary Gland Morphogenesis

et al. 2012

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“…A very small fraction of these focal lesions further progresses to invasive, but rarely metastatic mammary carcinomas 3, 4. These SV40‐induced mammary tumors exhibit either a basal‐like, morphologically differentiated phenotype (low‐grade tumors), or an undifferentiated phenotype (high‐grade tumors) featured by coexpression of epithelial and mesenchymal markers, respectively 12. In comparison to other oncoproteins, e.g.…”

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confidence: 68%

Low‐grade and high‐grade mammary carcinomas in WAP‐T transgenic mice are independent entities distinguished by Met expression

Otto

Gruner

Heinlein

et al. 2012

Intl Journal of Cancer

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Mammary carcinomas developing in SV40 transgenic WAP-T mice arise in two distinct histological phenotypes: as differentiated low-grade and undifferentiated high-grade tumors. We integrated different types of information such as histological grading, analysis of aCGH-based gene copy number and gene expression profiling to provide a comprehensive molecular description of mammary tumors in WAP-T mice. Applying a novel procedure for the correlation of gene copy number with gene expression on a global scale, we observed in tumor samples a global coherence between genotype and transcription. This coherence can be interpreted as a matched transcriptional regulation inherited from the cells of tumor origin and determined by the activity of cancer driver genes. Despite common recurrent genomic aberrations, e.g. gain of chr. 15 in most WAP-T tumors, loss of chr. 19 frequently occurs only in low-grade tumors. These tumors show features of ''basal-like'' epithelial differentiation, particularly expression of keratin 14. The high-grade tumors are clearly separated from the low-grade tumors by strong expression of the Met gene and by coexpression of epithelial (e.g. keratin 18) and mesenchymal (e.g. vimentin) markers. In high-grade tumors, the expression of the nonmutated Met protein is associated with Met-locus amplification and Met activity. The role of Met as a cancer driver gene is supported by the contribution of active Met signaling to motility and growth of mammary tumor-derived cells. Finally, we discuss the independent origin of low-and high-grade tumors from distinct cells of tumor origin, possibly luminal progenitors, distinguished by Met gene expression and Met signaling.Mouse models provide important tools for gaining insight into the biology of breast cancer (BC) and the relationship between histopathology and molecular profiles of cancer subtypes. 1,2 They narrow down the complexity of the disease and allow defining the influence of specific individual molecular factors and pathways on BC development and progression-information that is difficult to obtain from retrospective analyses of human tumor samples. Mammary epithelial

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“…Sections from all samples were stained with hematoxylin and eosin (H&E). In order to detect tumor cells in the lung, the lung was cut into serial sections (2 mm) and stained using anti-SV40 T-antigen (T-Ag) antibody (homemade rabbit antibody, R15, 1:20,000 14 ). Sections of all primary tumors (n 5 9-12 per group) were stained using antibodies against vimentin (rabbit monoclonal ab92547, Abcam, Cambridge, UK; 1:500), E-cadherin (rabbit monoclonal #3195, Cell Signaling Technology, Beverly, MA; 1:200), SV40 T-Ag (homemade), F4/80 (rat MCA497EL, AbD Serotec, Oxfordshire, UK; 1:100), CD45 (rat #103102, Biolegend, San Diego, CA; 1:600), Collagen-I (rabbit R1038, Acris; 1:200), alpha smooth muscle actin (rabbit ab5694, Abcam; 1:500) and ECF-L (M2 macrophages; goat #AF2446, R&D systems, Minneapolis, MN; 1:100) as well as the histochemical stainings with H&E, Masson-Goldner trichrome and Picrosirius red.…”

Section: Histologymentioning

confidence: 99%

Chemotherapy of WAP‐T mouse mammary carcinomas aggravates tumor phenotype and enhances tumor cell dissemination

Jannasch¹,

Wegwitz

Lenfert

et al. 2014

Intl Journal of Cancer

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In this study, the effects of the standard chemotherapy, cyclophosphamide/adriamycin/5-fluorouracil (CAF) on tumor growth, dissemination and recurrence after orthotopic implantation of murine G-2 cells were analyzed in the syngeneic immunocompetent whey acidic protein-T mouse model (Wegwitz et al., PLoS One 2010; 5:e12103; Schulze-Garg et al., Oncogene 2000; 19:1028-37). Single-dose CAF treatment reduced tumor size significantly, but was not able to eradicate all tumor cells, as recurrent tumor growth was observed 4 weeks after CAF treatment. Nine days after CAF treatment, residual tumors showed features of regressive alterations and were composed of mesenchymal-like tumor cells, infiltrating immune cells and some tumor-associated fibroblasts with an intense deposition of collagen. Recurrent tumors were characterized by coagulative necrosis and less tumor cell differentiation compared with untreated tumors, suggesting a more aggressive tumor phenotype. In support, tumor cell dissemination was strongly enhanced in mice that had developed recurrent tumors in comparison with untreated controls, although only few disseminated tumor cells could be detected in various organs 9 days after CAF application. In vitro experiments revealed that CAF treatment of G-2 cells eliminates the vast majority of epithelial tumor cells, whereas tumor cells with a mesenchymal phenotype survive. These results together with the in vivo findings suggest that tumor cells that underwent epithelial-mesenchymal transition and/or exhibit stem-cell-like properties are difficult to eliminate using one round of CAF chemotherapy. The model system described here provides a valuable tool for the characterization of the effects of chemotherapeutic regimens on recurrent tumor growth and on tumor cell dissemination, thereby enabling the development and preclinical evaluation of novel therapeutic strategies to target mammary carcinomas.Mammary carcinoma is the most frequent type of cancer in women in the Western world.1 About 25% of the patients develop distant metastases and ultimately die of breast cancer. 2 The prognosis of cancer patients is largely determined by the occurrence of distant metastases. Disseminated tumor cells (DTCs) in the bone marrow at diagnosis and during follow-up have been demonstrated in several early breast cancer studies to be of clinical relevance 3,4 and are used in patient risk assessment.The high incidence of breast cancer and the high mortality of the disease ask for better treatment options. However, a major problem in evaluating the effects of cancer therapy in preclinical settings results from the fact that the main parameter analyzed is tumor growth, and treatment effects on DTCs are not monitored. Especially, for treating mammary carcinomas, strategies to target DTCs and metastases are absolutely essential, as they are known to disseminate at an early stage. 5Most preclinical studies are performed using xenograft models implanting human tumor cells into immunocompromised mice, which only inadequately mimic tumor de...

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Tumorigenic WAP-T Mouse Mammary Carcinoma Cells: A Model for a Self-Reproducing Homeostatic Cancer Cell System

Cited by 25 publications

References 77 publications

Slug Controls Stem/Progenitor Cell Growth Dynamics during Mammary Gland Morphogenesis

Slug Controls Stem/Progenitor Cell Growth Dynamics during Mammary Gland Morphogenesis

Low‐grade and high‐grade mammary carcinomas in WAP‐T transgenic mice are independent entities distinguished by Met expression

Chemotherapy of WAP‐T mouse mammary carcinomas aggravates tumor phenotype and enhances tumor cell dissemination

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