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Patched1 deletion increases N-Myc protein stability as a mechanism of medulloblastoma initiation and progression

Oncogene volume 28, pages 1605–1615 (2009)Cite this article

Abstract

Medulloblastoma tumorigenesis caused by inactivating mutations in the PATCHED1 (PTCH1) gene is initiated by persistently activated Sonic Hedgehog (Shh) signaling in granule neuron precursors (GNPs) during the late stages of cerebellar development. Both normal cerebellar development and Shh-driven medulloblastoma tumorigenesis require N-Myc expression. However, the mechanisms by which N-Myc affects the stages of medulloblastoma initiation and progression are unknown. Here we used a mouse model of Ptch1 heterozygosity and medulloblastoma to show that increased N-Myc expression characterized the earliest selection of focal GNP hyperplasia destined for later tumor progression. Step-wise loss of Ptch1 expression, from tumor initiation to progression, led to incremental increases in N-Myc protein, rather than mRNA, expression. Increased N-Myc resulted in enhanced proliferation and death resistance of perinatal GNPs at tumor initiation. Sequential N-Myc protein phosphorylation at serine-62 and serine-62/threonine-58 characterized the early and late stages of medulloblastoma tumorigenesis, respectively. Shh pathway activation led to increased Myc protein stability and reduced expression of key regulatory factors. Taken together our data identify N-Myc protein stability as the result of loss of Ptch1, which distinguishes normal cerebellar development from medulloblastoma tumorigenesis.

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References

  • Arnold HK, Sears RC . (2006). Protein phosphatase 2A regulatory subunit B56alpha associates with c-myc and negatively regulates c-myc accumulation. Mol Cell Biol 26: 2832–2844.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Arnold HK, Sears RC . (2008). A tumor suppressor role for PP2A-B56alpha through negative regulation of c-Myc and other key oncoproteins. Cancer Metastasis Rev 27: 147–158.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Bahram F, von der Lehr N, Cetinkaya C, Larsson LG . (2000). c-Myc hot spot mutations in lymphomas result in inefficient ubiquitination and decreased proteasome-mediated turnover. Blood 95: 2104–2110.

    CAS  PubMed  Google Scholar 

  • Barnes EA, Heidtman KJ, Donoghue DJ . (2005). Constitutive activation of the shh–ptc1 pathway by a patched1 mutation identified in BCC. Oncogene 24: 902–915.

    Article  CAS  PubMed  Google Scholar 

  • Benassi B, Fanciulli M, Fiorentino F, Porrello A, Chiorino G, Loda M et al. (2006). c-Myc phosphorylation is required for cellular response to oxidative stress. Mol Cell 21: 509–519.

    Article  CAS  PubMed  Google Scholar 

  • Blackwood EM, Eisenman RN . (1991). Max: a helix-loop-helix zipper protein that forms a sequence-specific DNA-binding complex with Myc. Science 251: 1211–1217.

    Article  CAS  PubMed  Google Scholar 

  • Browd SR, Kenney AM, Gottfried ON, Yoon JW, Walterhouse D, Pedone CA et al. (2006). N-myc can substitute for insulin-like growth factor signaling in a mouse model of sonic hedgehog-induced medulloblastoma. Cancer Res 66: 2666–2672.

    Article  CAS  PubMed  Google Scholar 

  • Corcoran RB, Scott MP . (2001). A mouse model for medulloblastoma and basal cell nevus syndrome. J Neurooncol 53: 307–318.

    Article  CAS  PubMed  Google Scholar 

  • Corrales JD, Blaess S, Mahoney EM, Joyner AL . (2006). The level of sonic hedgehog signaling regulates the complexity of cerebellar foliation. Development 133: 1811–1821.

    Article  CAS  PubMed  Google Scholar 

  • Dahmane N, Ruiz i Altaba A . (1999). Sonic hedgehog regulates the growth and patterning of the cerebellum. Development 126: 3089–3100.

    PubMed  Google Scholar 

  • Dang CV . (1999). c-Myc target genes involved in cell growth, apoptosis, and metabolism. Mol Cell Biol 19: 1–11.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Eberhart CG, Kratz J, Wang Y, Summers K, Stearns D, Cohen K et al. (2004). Histopathological and molecular prognostic markers in medulloblastoma: c-myc, N-myc, TrkC, and anaplasia. J Neuropathol Exp Neurol 63: 441–449.

    Article  CAS  PubMed  Google Scholar 

  • Ellis T, Smyth I, Riley E, Graham S, Elliot K, Narang M et al. (2003). Patched 1 conditional null allele in mice. Genesis 36: 158–161.

    Article  CAS  PubMed  Google Scholar 

  • Evan GI, Wyllie AH, Gilbert CS, Littlewood TD, Land H, Brooks M et al. (1992). Induction of apoptosis in fibroblasts by c-myc protein. Cell 69: 119–128.

    Article  CAS  PubMed  Google Scholar 

  • Felsher DW . (2008). Oncogene addiction versus oncogene amnesia: perhaps more than just a bad habit? Cancer Res 68: 3081–3086; discussion 3086.

    Article  CAS  PubMed  Google Scholar 

  • Ferrer I, Soriano E, del Rio JA, Alcantara S, Auladell C . (1992). Cell death and removal in the cerebral cortex during development. Prog Neurobiol 39: 1–43.

    Article  CAS  PubMed  Google Scholar 

  • Fogarty MP, Kessler JD, Wechsler-Reya RJ . (2005). Morphing into cancer: the role of developmental signaling pathways in brain tumor formation. J Neurobiol 64: 458–475.

    Article  CAS  PubMed  Google Scholar 

  • Garson JA, Pemberton LF, Sheppard PW, Varndell IM, Coakham HB, Kemshead JT . (1989). N-myc gene expression and oncoprotein characterisation in medulloblastoma. Br J Cancer 59: 889–894.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Goodrich LV, Milenkovic L, Higgins KM, Scott MP . (1997). Altered neural cell fates and medulloblastoma in mouse patched mutants. Science 277: 1109–1113.

    Article  CAS  PubMed  Google Scholar 

  • Haber M, Bordow SB, Gilbert J, Madafiglio J, Kavallaris M, Marshall GM et al. (1999). Altered expression of the MYCN oncogene modulates MRP gene expression and response to cytotoxic drugs in neuroblastoma cells. Oncogene 18: 2777–2782.

    Article  CAS  PubMed  Google Scholar 

  • Hahn H, Wojnowski L, Specht K, Kappler R, Calzada-Wack J, Potter D et al. (2000). Patched target Igf2 is indispensable for the formation of medulloblastoma and rhabdomyosarcoma. J Biol Chem 275: 28341–28344.

    Article  CAS  PubMed  Google Scholar 

  • Hansford LM, Thomas WD, Keating JM, Burkhart CA, Peaston AE, Norris MD et al. (2004). Mechanisms of embryonal tumor initiation: distinct roles for MycN expression and MYCN amplification. Proc Natl Acad Sci USA 101: 12664–12669.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Hatton BA, Knoepfler PS, Kenney AM, Rowitch DH, de Alboran IM, Olson JM et al. (2006). N-myc is an essential downstream effector of Shh signaling during both normal and neoplastic cerebellar growth. Cancer Res 66: 8655–8661.

    Article  CAS  PubMed  Google Scholar 

  • Hatton BA, Villavicencio EH, Tsuchiya KD, Pritchard JI, Ditzler S, Pullar B et al. (2008). The Smo/Smo model: hedgehog-induced medulloblastoma with 90% incidence and leptomeningeal spread. Cancer Res 68: 1768–1776.

    Article  CAS  PubMed  Google Scholar 

  • Junttila MR, Puustinen P, Niemela M, Ahola R, Arnold H, Bottzauw T et al. (2007). CIP2A inhibits PP2A in human malignancies. Cell 130: 51–62.

    Article  CAS  PubMed  Google Scholar 

  • Kadin ME, Rubinstein LJ, Nelson JS . (1970). Neonatal cerebellar medulloblastoma originating from the fetal external granular layer. J Neuropathol Exp Neurol 29: 583–600.

    Article  CAS  PubMed  Google Scholar 

  • Kenney AM, Cole MD, Rowitch DH . (2003). Nmyc upregulation by sonic hedgehog signaling promotes proliferation in developing cerebellar granule neuron precursors. Development 130: 15–28.

    Article  CAS  PubMed  Google Scholar 

  • Kenney AM, Widlund HR, Rowitch DH . (2004). Hedgehog and PI-3 kinase signaling converge on Nmyc1 to promote cell cycle progression in cerebellar neuronal precursors. Development 131: 217–228.

    Article  CAS  PubMed  Google Scholar 

  • Kim JY, Nelson AL, Algon SA, Graves O, Sturla LM, Goumnerova LC et al. (2003). Medulloblastoma tumorigenesis diverges from cerebellar granule cell differentiation in patched heterozygous mice. Dev Biol 263: 50–66.

    Article  CAS  PubMed  Google Scholar 

  • Knoepfler PS, Kenney AM . (2006). Neural precursor cycling at sonic speed: N-Myc pedals, GSK-3 brakes. Cell Cycle 5: 47–52.

    Article  CAS  PubMed  Google Scholar 

  • Knudson Jr AG . (1971). Mutation and cancer: statistical study of retinoblastoma. Proc Natl Acad Sci USA 68: 820–823.

    Article  PubMed  PubMed Central  Google Scholar 

  • Koppel H, Lewis PD . (1983). Cell proliferation and DNA-synthesis in the external granular layer of the postnatal rat cerebellum: a quantitative study. Neuropathol Appl Neurobiol 9: 207–214.

    Article  CAS  PubMed  Google Scholar 

  • Lelievre V, Seksenyan A, Nobuta H, Yong WH, Chhith S, Niewiadomski P et al. (2008). Disruption of the PACAP gene promotes medulloblastoma in ptc1 mutant mice. Dev Biol 313: 359–370.

    Article  CAS  PubMed  Google Scholar 

  • Lossi L, Merighi A . (2003). In vivo cellular and molecular mechanisms of neuronal apoptosis in the mammalian CNS. Prog Neurobiol 69: 287–312.

    Article  CAS  PubMed  Google Scholar 

  • Lum L, Beachy PA . (2004). The Hedgehog response network: sensors, switches, and routers. Science 304: 1755–1759.

    Article  CAS  PubMed  Google Scholar 

  • Moriuchi S, Shimizu K, Miyao Y, Hayakawa T . (1996). An immunohistochemical analysis of medulloblastoma and PNET with emphasis on N-myc protein expression. Anticancer Res 16: 2687–2692.

    CAS  PubMed  Google Scholar 

  • Murone M, Rosenthal A, de Sauvage FJ . (1999). Sonic hedgehog signaling by the patched-smoothened receptor complex. Curr Biol 9: 76–84.

    Article  CAS  PubMed  Google Scholar 

  • Nesbit CE, Tersak JM, Prochownik EV . (1999). MYC oncogenes and human neoplastic disease. Oncogene 18: 3004–3016.

    Article  CAS  PubMed  Google Scholar 

  • Oliver TG, Grasfeder LL, Carroll AL, Kaiser C, Gillingham CL, Lin SM et al. (2003). Transcriptional profiling of the Sonic hedgehog response: a critical role for N-myc in proliferation of neuronal precursors. Proc Natl Acad Sci USA 100: 7331–7336.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Oliver TG, Read TA, Kessler JD, Mehmeti A, Wells JF, Huynh TT et al. (2005). Loss of patched and disruption of granule cell development in a pre-neoplastic stage of medulloblastoma. Development 132: 2425–2439.

    Article  CAS  PubMed  Google Scholar 

  • Pizzo PA, Poplack DG . (2002). Principles and Practice of Pediatric Oncology. Lipincott Williams and Wilkins: Philadelphia, PA.

    Google Scholar 

  • Pomeroy SL, Tamayo P, Gaasenbeek M, Sturla LM, Angelo M, McLaughlin ME et al. (2002). Prediction of central nervous system embryonal tumour outcome based on gene expression. Nature 415: 436–442.

    Article  CAS  PubMed  Google Scholar 

  • Popov N, Wanzel M, Madiredjo M, Zhang D, Beijersbergen R, Bernards R et al. (2007). The ubiquitin-specific protease USP28 is required for MYC stability. Nat Cell Biol 9: 765–774.

    Article  CAS  PubMed  Google Scholar 

  • Pritchard JI, Olson JM . (2008). Methylation of PTCH1, the Patched-1 gene, in a panel of primary medulloblastomas. Cancer Genet Cytogenet 180: 47–50.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Romer JT, Kimura H, Magdaleno S, Sasai K, Fuller C, Baines H et al. (2004). Suppression of the Shh pathway using a small molecule inhibitor eliminates medulloblastoma in Ptc1(+/−)p53(−/−) mice. Cancer Cell 6: 229–240.

    Article  CAS  PubMed  Google Scholar 

  • Ruiz i Altaba A, Stecca B, Sanchez P . (2004). Hedgehog—Gli signaling in brain tumors: stem cells and paradevelopmental programs in cancer. Cancer Lett 204: 145–157.

    Article  CAS  PubMed  Google Scholar 

  • Sasai K, Romer JT, Lee Y, Finkelstein D, Fuller C, McKinnon PJ et al. (2006). Shh pathway activity is down-regulated in cultured medulloblastoma cells: implications for preclinical studies. Cancer Res 66: 4215–4222.

    Article  CAS  PubMed  Google Scholar 

  • Sears R, Nuckolls F, Haura E, Taya Y, Tamai K, Nevins JR . (2000). Multiple Ras-dependent phosphorylation pathways regulate Myc protein stability. Genes Dev 14: 2501–2514.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Sears RC . (2004). The life cycle of C-myc: from synthesis to degradation. Cell Cycle 3: 1133–1137.

    Article  CAS  PubMed  Google Scholar 

  • Sjostrom SK, Finn G, Hahn WC, Rowitch DH, Kenney AM . (2005). The Cdk1 complex plays a prime role in regulating N-myc phosphorylation and turnover in neural precursors. Dev Cell 9: 327–338.

    Article  CAS  PubMed  Google Scholar 

  • Smith DP, Bath ML, Metcalf D, Harris AW, Cory S . (2006). MYC levels govern hematopoietic tumor type and latency in transgenic mice. Blood 108: 653–661.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Tronche F, Kellendonk C, Kretz O, Gass P, Anlag K, Orban PC et al. (1999). Disruption of the glucocorticoid receptor gene in the nervous system results in reduced anxiety. Nat Genet 23: 99–103.

    Article  CAS  PubMed  Google Scholar 

  • Wechsler-Reya RJ . (2003). Analysis of gene expression in the normal and malignant cerebellum. Recent Prog Horm Res 58: 227–248.

    Article  CAS  PubMed  Google Scholar 

  • Wechsler-Reya RJ, Scott MP . (1999). Control of neuronal precursor proliferation in the cerebellum by Sonic Hedgehog. Neuron 22: 103–114.

    Article  CAS  PubMed  Google Scholar 

  • Wetmore C, Eberhart DE, Curran T . (2000). The normal patched allele is expressed in medulloblastomas from mice with heterozygous germ-line mutation of patched. Cancer Res 60: 2239–2246.

    CAS  PubMed  Google Scholar 

  • Yeh E, Cunningham M, Arnold H, Chasse D, Monteith T, Ivaldi G et al. (2004). A signalling pathway controlling c-Myc degradation that impacts oncogenic transformation of human cells. Nat Cell Biol 6: 308–318.

    Article  CAS  PubMed  Google Scholar 

  • Zakhary RK, Keles GE, Aldape K, Berger MS . (2001) Medulloblastoma and primitive neuroectodermal tumors. In: Kaye AH, Law ER (eds). Brain Tumours: An Encyclopedic Approach. Churchill Livingstone: London, pp 605–615.

    Google Scholar 

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Acknowledgements

This work was supported by funds from the Cure For Life Foundation (WT), the Cure Cancer Australia Foundation (WT), the National Health and Medical Research Council (Australia; GMM, MDN, BW and MH), the Cancer Institute New South Wales (Australia; GMM, MDN and MH) and John Trivett Foundation (TE). The Children's Cancer Institute Australia for Medical Research is affiliated with the University of New South Wales and Sydney Children's Hospital.

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  1. W D Thomas and J Chen: These authors contributed equally to this work.

Authors and Affiliations

  1. Children's Cancer Institute Australia for Medical Research, Randwick, New South Wales, Australia

    W D Thomas, J Chen, Y R Gao, B Cheung, J Koach, E Sekyere, M D Norris, M Haber & G M Marshall

  2. Institute for Molecular Bioscience, University of Queensland, Brisbane, Queensland, Australia

    T Ellis & B Wainwright

  3. Centre for Children's Cancer and Blood Disorders, Sydney Children's Hospital, Randwick, New South Wales, Australia

    G M Marshall

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  1. W D Thomas
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Correspondence to G M Marshall.

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Supplementary Information accompanies the paper on the Oncogene website (http://www.nature.com/onc)

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Thomas, W., Chen, J., Gao, Y. et al. Patched1 deletion increases N-Myc protein stability as a mechanism of medulloblastoma initiation and progression. Oncogene 28, 1605–1615 (2009). https://doi.org/10.1038/onc.2009.3

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