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Development of mammary hyperplasia, dysplasia, and invasive ductal carcinoma in transgenic mice expressing the 8p11 amplicon oncogene NSD3

  • Preclinical study
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Abstract

Purpose

NSD3 has been implicated as a candidate driver oncogene from the 8p11-p12 locus, and we have previously published evidence for its amplification and overexpression in human breast cancer. This aim of this study was to further characterize the transforming function of NSD3 in vivo.

Methods

We generated a transgenic mouse model in which NSD3 gene expression was driven by the MMTV promoter and expressed in mammary epithelium of FVB mice. Mammary glands were fixed and whole mounts were stained with carmine to visualize gland structure. Mammary tumors were formalin-fixed, and paraffin embedded (FFPE) tumors were stained with hematoxylin and eosin.

Results

Pups born to transgenic females were significantly underdeveloped compared to pups born to WT females due to a lactation defect in transgenic female mice. Whole mount analysis of the mammary glands of transgenic female mice revealed a profound defect in functional differentiation of mammary gland alveoli that resulted in the lactation defect. We followed parous and virgin NSD3 transgenic and control mice to 50 weeks of age and observed that several NSD3 parous females developed mammary tumors. Whole mount analysis of the mammary glands of tumor-bearing mice revealed numerous areas of mammary hyperplasia and ductal dysplasia. Histological analysis showed that mammary tumors were high-grade ductal carcinomas, and lesions present in other mammary glands exhibited features of alveolar hyperplasia, ductal dysplasia, and carcinoma in situ.

Conclusions

Our results are consistent with our previous studies and demonstrate that NSD3 is a transforming breast cancer oncogene.

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Acknowledgements

We would like to specifically thank Michael J. Kern, Ph.D. of the Gene Function Core at the Medical University of South Carolina and all of the technicians for their assistance in generating the NSD3 transgenic mice. We would also like to thank Kiwana Gibbs and Yuan Shao, Ph.D. of the Biorepository and Tissue Analysis Shared Resource at the Medical University of South Carolina for providing histological sections and staining of all mammary glands and tumors.

Authors’ contribution

Conception and design of the study was conducted by SE. Transgenic mice were handled by ES for ear tagging/tail snips, euthanizing, and harvesting of all mammary glands for whole mount preparation. Genotyping and transgenic mouse record keeping for the NSD3 colony was performed by BI. Both ES and BI performed carmine staining of whole mounts, cultured tumor-derived mammary cells, and performed western blotting on lysates obtained from NSD3-derived mammary tumor cells. AR organized mammary glands and generated images for scoring hyperplastic lesions seen in whole mounts. LS is the breast cancer pathologist for the Hollings Cancer Center and along with JM performed histopathologic analysis of mammary lesions and tumors. The manuscript was written by SE, BI, ES, AR, and LS.

Funding

This study was funded by the 2RO1CA100724, NCI 2P30CA138313, and the Chalsty Breast Cancer Research Fund 1K12CA457688-O1A1.

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Author notes

    Authors and Affiliations

    1. Department of Pathology and Laboratory Medicine, Hollings Cancer Center, Medical University of South Carolina, 68 President St, Charleston, SC, 29425, USA

      Brittany Turner-Ivey, Ericka L. Smith, Alex C. Rutkovsky, Laura S. Spruill, Jamie N. Mills & Stephen P. Ethier

    2. Hollings Cancer Center, Medical University of South Carolina, Charleston, SC, USA

      Laura S. Spruill

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    1. Brittany Turner-Ivey
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    Correspondence to Stephen P. Ethier.

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    The authors declare that they have no conflict of interest.

    Ethical approval

    All applicable international, national, and/or institutional guidelines for the care and use of animals were followed. The Institutional Animal Care and Use Committee (IACUC) at the Medical University of SC (MUSC) approved all animal experiments, AR#3397. Male and female WT FVB/N mice were bred under typical conditions with unlimited access to food and water. Animals were euthanized per IACUC approved protocol during all experiments. This article does not contain any studies with human participants performed by any of the authors.

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    Brittany Turner-Ivey and Ericka L. Smith have contributed equally to this work.

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    Turner-Ivey, B., Smith, E.L., Rutkovsky, A.C. et al. Development of mammary hyperplasia, dysplasia, and invasive ductal carcinoma in transgenic mice expressing the 8p11 amplicon oncogene NSD3. Breast Cancer Res Treat 164, 349–358 (2017). https://doi.org/10.1007/s10549-017-4258-9

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    • DOI: https://doi.org/10.1007/s10549-017-4258-9

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