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A requirement for Nedd9 in luminal progenitor cells prior to mammary tumorigenesis in MMTV-HER2/ErbB2 mice

Oncogene volume 33, pages 411–420 (2014)Cite this article

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Abstract

Overexpression of the NEDD9/HEF1/Cas-L scaffolding protein is frequent, and drives invasion and metastasis in breast, head and neck, colorectal, melanoma, lung and other types of cancer. We have examined the consequences of genetic ablation of Nedd9 in the MMTV-HER2/ERBB2/neu mouse mammary tumor model. Unexpectedly, we found that only a limited effect on metastasis in MMTV-neu;Nedd9−/− mice compared with MMTV-neu;Nedd9+/+ mice, but instead a dramatic reduction in tumor incidence (18 versus 80%), and a significantly increased latency until tumor appearance. Orthotopic reinjection and tail-vein injection of cells arising from tumors, coupled with in vivo analysis, indicated tumors arising in MMTV-neu;Nedd9−/− mice had undergone mutational selection that overcame the initial requirement for Nedd9. To better understand the defects in early tumor growth, we compared mammary progenitor cell pools from MMTV-neu;Nedd9−/− versus MMTV-neu;Nedd9+/+ mice. The MMTV-neu;Nedd9−/− genotype selectively reduced both the number and colony-forming potential of mammary luminal epithelial progenitor cells, while not affecting basal epithelial progenitors. MMTV-neu;Nedd9−/− mammospheres had striking defects in morphology and cell polarity. All of these defects were seen predominantly in the context of the HER2/neu oncogene, and were not associated with randomization of the plane of mitotic division, but rather with depressed expression the cell attachment protein FAK, accompanied by increased sensitivity to small molecule inhibitors of FAK and SRC. Surprisingly, in spite of these significant differences, only minimal changes were observed in the gene expression profile of Nedd9−/− mice, indicating critical Nedd9-dependent differences in cell growth properties were mediated via post-transcriptional regulation of cell signaling. Coupled with emerging data indicating a role for NEDD9 in progenitor cell populations during the morphogenesis of other tissues, these results indicate a functional requirement for NEDD9 in the growth of mammary cancer progenitor cells.

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Acknowledgements

We are grateful to Emmanuelle Nicolas (Genotyping and Real-Time-PCR Facility of FCCC), Jianming Pei (Cytogenetics and Chromosome Microarray Analysis Facility of FCCC) and Yue-Sheng Liu (Expression Microarray Facility of FCCC), Anthony Lerro and Jackie Valvardi (Laboratory Animal Facility of FCCC), James Oesterling (Cell Sorting Facility of FCCC), Jin Fang (Cell Culture Facility of FCCC), Catherine Renner (Histopathology Facility), Marina Boushra (from Cornell University), Christiaan Honig (from Germantown Academy) and Nicolas G Day (from St John’s College) for technical help. We particularly acknowledge Edna Cukierman for expert advice with analysis of microscopic images. R01s CA63366 and CA113342, a U54 CA149147 subcontract, and Pennsylvania Tobacco Settlement funding (to EAG); and NIH core Grant CA06927 and the Pew Charitable Fund (to Fox Chase Cancer Center) supported this work. JLL was supported by NIH T32 CA009035 and F32 CA150553; EI by the American Associates, Ben-Gurion University of the Negev; EP by the Fox Chase Cancer Center Undergraduate Student Summer Fellowship Program. MFO was supported by R21 LM009382.

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  1. E Izumchenko

    Present address: 6Current address: Department of Biochemistry and Molecular Biology, University of Florida, Gainesville, FL, USA.,

Authors and Affiliations

  1. Fox Chase Cancer Center, Institute of Cancer Research, Philadelphia, PA, USA

    J L Little, V Serzhanova, E Izumchenko, B L Egleston, A J Klein-Szanto, M Shubina & E A Golemis

  2. Department of Bioengineering, University of Pittsburgh, Pittsburgh, PA, USA

    E Parise

  3. Department of Biology, Bryn Mawr College, Bryn Mawr, PA, USA

    J L Little & G Loudon

  4. Department of Hematology and Oncology, Graduate School of Medicine, University of Tokyo, Tokyo, Japan

    S Seo & M Kurokawa

  5. Departments of Oncology and Health Science Informatics, Johns Hopkins University, Baltimore, MD, USA

    M F Ochs

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Little, J., Serzhanova, V., Izumchenko, E. et al. A requirement for Nedd9 in luminal progenitor cells prior to mammary tumorigenesis in MMTV-HER2/ErbB2 mice. Oncogene 33, 411–420 (2014). https://doi.org/10.1038/onc.2012.607

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