miR-380-5p represses p53 to control cellular survival and is associated with poor outcome in MYCN-amplified neuroblastoma

Abstract

Inactivation of the p53 tumor suppressor pathway allows cell survival in times of stress and occurs in many human cancers; however, normal embryonic stem cells and some cancers such as neuroblastoma maintain wild-type human TP53 and mouse Trp53 (referred to collectively as p53 herein). Here we describe a miRNA, miR-380-5p, that represses p53 expression via a conserved sequence in the p53 3′ untranslated region (UTR). miR-380-5p is highly expressed in mouse embryonic stem cells and neuroblastomas, and high expression correlates with poor outcome in neuroblastomas with neuroblastoma derived v-myc myelocytomatosis viral-related oncogene (MYCN) amplification. miR-380 overexpression cooperates with activated HRAS oncoprotein to transform primary cells, block oncogene-induced senescence and form tumors in mice. Conversely, inhibition of endogenous miR-380-5p in embryonic stem or neuroblastoma cells results in induction of p53, and extensive apoptotic cell death. In vivo delivery of a miR-380-5p antagonist decreases tumor size in an orthotopic mouse model of neuroblastoma. We demonstrate a new mechanism of p53 regulation in cancer and stem cells and uncover a potential therapeutic target for neuroblastoma.

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Figure 1: The p53 3′ UTR contains binding sites for miR-380-5p, a developmentally restricted miRNA.
Figure 2: miR-380-5p is required for ES cell survival.
Figure 3: miR-380-5p targets p53 and decreases cell death after genotoxic stress.
Figure 4: miR-380 prevents oncogene-induced senescence and increases tumor incidence in a mouse mammary cancer model.
Figure 5: miR-380-5p is expressed in mouse and human neuroblastoma and is associated with poor outcome in subjects with MYCN amplification.
Figure 6: Treatment with miR-380-5p antagonist induces p53-dependent cell death in neuroblastoma cells and decreases tumor growth in vivo.

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Acknowledgements

We gratefully thank J.M. Bishop, N.K. Hayward and R.L. Sutherland for their support of this project, the Children's Oncology Group and M. Grimmer (University of California–San Francisco) for providing tumor samples, D. Lynch and J. Brugge (Harvard Medical School) for MCF10A cells expressing the ecotropic receptor, R. Jaenisch (Whitehead Institute) for Trp53−/− ES cells and S. Lowe (Cold Spring Harbor) for the p53 shRNA retrovirus. TH-MYCN transgenic mice were from W. Weiss (University of California–San Francisco). This work was supported by grants from the US National Institutes of Health: P50-CA58207, K08-CA104032 and 1R01CA136717 (to A.G.), 5R01DC005991 (to N.L.), R01CA102321, R01NS055750 and P01CA081403 (to W.W.) and K08NS48118 (to R.B.); the Susan G. Komen Foundation; the University of California–San Francisco Program for Breakthrough Biomedical Research (to A.G.); the G.W. Hooper Foundation; the Australian National Health and Medical Research Council (to T.P., M.H., M.D.N. and A. Swarbrick) and the Cancer Institute New South Wales (M.H. and M.D.N.). A. Swarbrick is a recipient of a Cancer Institute New South Wales early career development fellowship, and R.L.J. received a US National Science Foundation fellowship. A.G. is a V-Foundation Scholar, A. Shaw is a Cancer Institute New South Wales Research Scholar and Y.P. is supported by an Australian Postgraduate Award from the Australian National Health and Medical Research Council.

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S.L.W., A. Swarbrick and A.G. conceived and designed the experiments, discussed the results and wrote the manuscript. A. Swarbrick, S.L.W., A. Shaw, Y.P., A.N., A.G., R.L.J., C.S.S., C.S.H., P.L., A.B., N.H., Y.C. and L.L. performed experiments. L.J.A. and M.D.N. performed statistical analysis of the human neuroblastoma data set. A.C. provided human samples and clinical data, and E.G.M. provided anti-miRs for in vivo studies. M.H., T.P., W.W., N.L. and C.S.S. supervised experiments or experimental design.

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Correspondence to Alexander Swarbrick or Andrei Goga.

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E.G.M. is an employee and shareholder of Regulus Therapeutics.

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Swarbrick, A., Woods, S., Shaw, A. et al. miR-380-5p represses p53 to control cellular survival and is associated with poor outcome in MYCN-amplified neuroblastoma. Nat Med 16, 1134–1140 (2010). https://doi.org/10.1038/nm.2227

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