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The miR-106b-25 cluster mediates breast tumor initiation through activation of NOTCH1 via direct repression of NEDD4L

Oncogene volume 37, pages 3879–3893 (2018)Cite this article

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Abstract

Tumor-initiating cells (TIC) represent a subset of tumor cells with increased self-renewal capability. TICs display resistance to frontline cancer treatment and retain the ability to repopulate a tumor after therapy, leading to cancer relapse. NOTCH signaling has been identified as an important driver of the TIC population, yet mechanisms governing regulation of this pathway in cancer remain to be fully elucidated. Here we identify a novel mechanism of NOTCH regulation and TIC induction in breast cancer via the miR-106b-25 miRNA cluster. We show that the miR-106b-25 cluster upregulates NOTCH1 in multiple breast cancer cell lines, representing both estrogen receptor (ER+) and triple negative breast cancer (TNBC) through direct repression of the E3 ubiquitin ligase, NEDD4L. We further show that upregulation of NOTCH1 is necessary for TIC induction downstream of miR-106b-25 in both ER + and TNBC breast cancer cells, and that re-expression of NEDD4L is sufficient to reverse miR106b-25-mediated NOTCH1 upregulation and TIC induction. Importantly, we demonstrate a significant positive correlation between miR-106b-25 and NOTCH1 protein, yet a significant inverse correlation between miR-106b-25 and NEDD4L mRNA in human breast cancer, suggesting a critical role for the miR106b-25/NEDD4L/NOTCH1 axis in the disease. Further, we show for the first time that NEDD4L expression alone is significantly associated with a better relapse-free prognosis for breast cancer patients. These data expand our knowledge of the mechanisms underlying NOTCH activation and TIC induction in breast cancer, and may provide new avenues for the development of therapies targeting this resistant subset of tumor cells.

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Acknowledgements

The authors thank Paul Jedlicka for the generous gift of mice for limiting dilution experiments. We also want to thank the University of Colorado Cancer center flow cytometry shared resource for performing flow cytometry experiments needed for this work (P30CA046934), the Functional Genomics Facility for shRNA constructs, and The Cancer Genome Atlas (TCGA) for use of their breast cancer patient data set. This work was supported by R01CA095277 (HLF), R01CA117907 (JME), the METAvivor Foundation (HLF), the Department of Defense Breast Cancer Research Program W81XWH-10-1-0296 (ALG), the Ruth L. Kirschstein National Service Research Award (F32) 1F32CA199716-01 (ALG), the Ruth L. Kirschstein National Service Research Award (F31) 5F31CA210622-02 (MUJO), the NCI fellow transition award F99 CA223023 (MUJO), UC Denver AMC Molecular Biology Program T32 training grant T32 GM008730 (CGT), NIH-RO1 Diversity Supplement to R01-CA157790 (CGT), the UNCF/MERCK Graduate Fellowship (CGT), and the T32 Cancer Biology training grant CA190216-1 A1 (CGT).

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  1. These authors contributed equally: AL Guarnieri, CG Towers.

Authors and Affiliations

  1. Program in Molecular Biology, University of Colorado Anschutz Medical Campus, Aurora, CO, 80045, USA

    A. L. Guarnieri, C. G. Towers, D. J. Drasin, R. L. Vartuli, E. S. Linklater & H. L. Ford

  2. Department of Pharmacology, University of Colorado Anschutz Medical Campus, Aurora, CO, 80045, USA

    A. L. Guarnieri, C. G. Towers, M. U. J. Oliphant, Z. Andrysik, T. J. Hotz, R. L. Vartuli, A. Pandey, S. Khanal, J. M. Espinosa & H. L. Ford

  3. Integrated Physiology Program, University of Colorado Anschutz Medical Campus, Aurora, CO, 80045, USA

    M. U. J. Oliphant & H. L. Ford

  4. Linda Crnic Institute for Down Syndrome, University of Colorado Anschutz Medical Campus, Aurora, CO, 80045, USA

    Z. Andrysik & J. M. Espinosa

  5. Department of Molecular, Cellular and Developmental Biology, University of Colorado at Boulder, Boulder, CO, 80309, USA

    J. M. Espinosa

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Guarnieri, A.L., Towers, C.G., Drasin, D.J. et al. The miR-106b-25 cluster mediates breast tumor initiation through activation of NOTCH1 via direct repression of NEDD4L. Oncogene 37, 3879–3893 (2018). https://doi.org/10.1038/s41388-018-0239-7

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