Nos2 Inactivation Promotes the Development of Medulloblastoma in Mice by Deregulation of Gap43–Dependent Granule Cell Precursor Migration
Medulloblastoma is the most common malignant brain tumor in children. A subset of medulloblastoma originates from granule cell precursors (GCPs) of the developing cerebellum and demonstrates aberrant hedgehog signaling, typically due to inactivating mutations in the receptor PTCH1, a pathomechanism recapitulated in Ptch1+/− mice. As nitric oxide may regulate GCP proliferation and differentiation, we crossed Ptch1+/− mice with mice lacking inducible nitric oxide synthase (Nos2) to investigate a possible influence on tumorigenesis. We observed a two-fold higher medulloblastoma rate in Ptch1+/− Nos2−/− mice compared to Ptch1+/− Nos2+/+ mice. To identify the molecular mechanisms underlying this finding, we performed gene expression profiling of medulloblastomas from both genotypes, as well as normal cerebellar tissue samples of different developmental stages and genotypes. Downregulation of hedgehog target genes was observed in postnatal cerebellum from Ptch1+/+ Nos2−/− mice but not from Ptch1+/− Nos2−/− mice. The most consistent effect of Nos2 deficiency was downregulation of growth-associated protein 43 (Gap43). Functional studies in neuronal progenitor cells demonstrated nitric oxide dependence of Gap43 expression and impaired migration upon Gap43 knock-down. Both effects were confirmed in situ by immunofluorescence analyses on tissue sections of the developing cerebellum. Finally, the number of proliferating GCPs at the cerebellar periphery was decreased in Ptch1+/+ Nos2−/− mice but increased in Ptch1+/− Nos2−/− mice relative to Ptch1+/− Nos2+/+ mice. Taken together, these results indicate that Nos2 deficiency promotes medulloblastoma development in Ptch1+/− mice through retention of proliferating GCPs in the external granular layer due to reduced Gap43 expression. This study illustrates a new role of nitric oxide signaling in cerebellar development and demonstrates that the localization of pre-neoplastic cells during morphogenesis is crucial for their malignant progression.
Vyšlo v časopise:
Nos2 Inactivation Promotes the Development of Medulloblastoma in Mice by Deregulation of Gap43–Dependent Granule Cell Precursor Migration. PLoS Genet 8(3): e32767. doi:10.1371/journal.pgen.1002572
Kategorie:
Research Article
prolekare.web.journal.doi_sk:
https://doi.org/10.1371/journal.pgen.1002572
Souhrn
Medulloblastoma is the most common malignant brain tumor in children. A subset of medulloblastoma originates from granule cell precursors (GCPs) of the developing cerebellum and demonstrates aberrant hedgehog signaling, typically due to inactivating mutations in the receptor PTCH1, a pathomechanism recapitulated in Ptch1+/− mice. As nitric oxide may regulate GCP proliferation and differentiation, we crossed Ptch1+/− mice with mice lacking inducible nitric oxide synthase (Nos2) to investigate a possible influence on tumorigenesis. We observed a two-fold higher medulloblastoma rate in Ptch1+/− Nos2−/− mice compared to Ptch1+/− Nos2+/+ mice. To identify the molecular mechanisms underlying this finding, we performed gene expression profiling of medulloblastomas from both genotypes, as well as normal cerebellar tissue samples of different developmental stages and genotypes. Downregulation of hedgehog target genes was observed in postnatal cerebellum from Ptch1+/+ Nos2−/− mice but not from Ptch1+/− Nos2−/− mice. The most consistent effect of Nos2 deficiency was downregulation of growth-associated protein 43 (Gap43). Functional studies in neuronal progenitor cells demonstrated nitric oxide dependence of Gap43 expression and impaired migration upon Gap43 knock-down. Both effects were confirmed in situ by immunofluorescence analyses on tissue sections of the developing cerebellum. Finally, the number of proliferating GCPs at the cerebellar periphery was decreased in Ptch1+/+ Nos2−/− mice but increased in Ptch1+/− Nos2−/− mice relative to Ptch1+/− Nos2+/+ mice. Taken together, these results indicate that Nos2 deficiency promotes medulloblastoma development in Ptch1+/− mice through retention of proliferating GCPs in the external granular layer due to reduced Gap43 expression. This study illustrates a new role of nitric oxide signaling in cerebellar development and demonstrates that the localization of pre-neoplastic cells during morphogenesis is crucial for their malignant progression.
Zdroje
1. EllisonDWKocakMDaltonJMegahedHLusherME 2011 Definition of disease-risk stratification groups in childhood medulloblastoma using combined clinical, pathologic, and molecular variables. J Clin Oncol 29 1400 1407
2. MulhernRKMerchantTEGajjarAReddickWEKunLE 2004 Late neurocognitive sequelae in survivors of brain tumours in childhood. Lancet Oncol 5 399 408
3. NorthcottPAKorshunovAWittHHielscherTEberhartCG 2011 Medulloblastoma comprises four distinct molecular variants. J Clin Oncol 29 1408 1414
4. KoolMKosterJBuntJHasseltNELakemanA 2008 Integrated genomics identifies five medulloblastoma subtypes with distinct genetic profiles, pathway signatures and clinicopathological features. PLoS ONE 3 e3088 doi:10.1371/journal.pone.0003088
5. SchullerUHeineVMMaoJKhoATDillonAK 2008 Acquisition of granule neuron precursor identity is a critical determinant of progenitor cell competence to form Shh-induced medulloblastoma. Cancer Cell 14 123 134
6. MillenKJGleesonJG 2008 Cerebellar development and disease. Curr Opin Neurobiol 18 12 19
7. Wechsler-ReyaRJScottMP 1999 Control of neuronal precursor proliferation in the cerebellum by Sonic Hedgehog. Neuron 22 103 114
8. CohenMMJr 2003 The hedgehog signaling network. Am J Med Genet A 123A 5 28
9. KenneyAMColeMDRowitchDH 2003 Nmyc upregulation by sonic hedgehog signaling promotes proliferation in developing cerebellar granule neuron precursors. Development 130 15 28
10. LeeEYJiHOuyangZZhouBMaW 2010 Hedgehog pathway-regulated gene networks in cerebellum development and tumorigenesis. Proc Natl Acad Sci U S A 107 9736 9741
11. PogorilerJMillenKUtsetMDuW 2006 Loss of cyclin D1 impairs cerebellar development and suppresses medulloblastoma formation. Development 133 3929 3937
12. KesslerJDHasegawaHBrunSNEmmeneggerBAYangZJ 2009 N-myc alters the fate of preneoplastic cells in a mouse model of medulloblastoma. Genes Dev 23 157 170
13. LumLBeachyPA 2004 The Hedgehog response network: sensors, switches, and routers. Science 304 1755 1759
14. EspinosaJSLuoL 2008 Timing neurogenesis and differentiation: insights from quantitative clonal analyses of cerebellar granule cells. J Neurosci 28 2301 2312
15. ten DonkelaarHJLammensMWesselingPThijssenHORenierWO 2003 Development and developmental disorders of the human cerebellum. J Neurol 250 1025 1036
16. ChoiYBorghesaniPRChanJASegalRA 2005 Migration from a mitogenic niche promotes cell-cycle exit. J Neurosci 25 10437 10445
17. PietschTWahaAKochAKrausJAlbrechtS 1997 Medulloblastomas of the desmoplastic variant carry mutations of the human homologue of Drosophila patched. Cancer Res 57 2085 2088
18. WolterMReifenbergerJSommerCRuzickaTReifenbergerG 1997 Mutations in the human homologue of the Drosophila segment polarity gene patched (PTCH) in sporadic basal cell carcinomas of the skin and primitive neuroectodermal tumors of the central nervous system. Cancer Res 57 2581 2585
19. JohnsonRLRothmanALXieJGoodrichLVBareJW 1996 Human homolog of patched, a candidate gene for the basal cell nevus syndrome. Science 272 1668 1671
20. GoodrichLVMilenkovicLHigginsKMScottMP 1997 Altered neural cell fates and medulloblastoma in mouse patched mutants. Science 277 1109 1113
21. BriggsKJCorcoran-SchwartzIMZhangWHarckeTDevereuxWL 2008 Cooperation between the Hic1 and Ptch1 tumor suppressors in medulloblastoma. Genes Dev 22 770 785
22. UzielTZindyFXieSLeeYForgetA 2005 The tumor suppressors Ink4c and p53 collaborate independently with Patched to suppress medulloblastoma formation. Genes Dev 19 2656 2667
23. MoncadaS 1997 Nitric oxide in the vasculature: physiology and pathophysiology. Ann N Y Acad Sci 811 60 67; discussion 67–69
24. ArancioOKieblerMLeeCJLev-RamVTsienRY 1996 Nitric oxide acts directly in the presynaptic neuron to produce long-term potentiation in cultured hippocampal neurons. Cell 87 1025 1035
25. NathanCXieQW 1994 Nitric oxide synthases: roles, tolls, and controls. Cell 78 915 918
26. KleinertHSchwarzPMForstermannU 2003 Regulation of the expression of inducible nitric oxide synthase. Biol Chem 384 1343 1364
27. FukumuraDKashiwagiSJainRK 2006 The role of nitric oxide in tumour progression. Nat Rev Cancer 6 521 534
28. YamaguchiKSaitoHOroSTatebeSIkeguchiM 2005 Expression of inducible nitric oxide synthase is significantly correlated with expression of vascular endothelial growth factor and dendritic cell infiltration in patients with advanced gastric carcinoma. Oncology 68 471 478
29. HussainSPTriversGEHofsethLJHePShaikhI 2004 Nitric oxide, a mediator of inflammation, suppresses tumorigenesis. Cancer Res 64 6849 6853
30. ManderPBorutaiteVMoncadaSBrownGC 2005 Nitric oxide from inflammatory-activated glia synergizes with hypoxia to induce neuronal death. J Neurosci Res 79 208 215
31. CharlesNOzawaTSquatritoMBleauAMBrennanCW 2010 Perivascular nitric oxide activates notch signaling and promotes stem-like character in PDGF-induced glioma cells. Cell Stem Cell 6 141 152
32. SatoASekineYSarutaCNishibeHMoritaN 2008 Cerebellar development transcriptome database (CDT-DB): profiling of spatio-temporal gene expression during the postnatal development of mouse cerebellum. Neural Netw 21 1056 1069
33. JuradoSSanchez-PrietoJTorresM 2004 Elements of the nitric oxide/cGMP pathway expressed in cerebellar granule cells: biochemical and functional characterisation. Neurochem Int 45 833 843
34. JeskoHChalimoniukMStrosznajderJB 2003 Activation of constitutive nitric oxide synthase(s) and absence of inducible isoform in aged rat brain. Neurochem Int 42 315 322
35. PackerMAStasivYBenraissAChmielnickiEGrinbergA 2003 Nitric oxide negatively regulates mammalian adult neurogenesis. Proc Natl Acad Sci U S A 100 9566 9571
36. CianiECalvaneseVCrochemoreCBartesaghiRContestabileA 2006 Proliferation of cerebellar precursor cells is negatively regulated by nitric oxide in newborn rat. J Cell Sci 119 3161 3170
37. CianiESeveriSContestabileABartesaghiR 2004 Nitric oxide negatively regulates proliferation and promotes neuronal differentiation through N-Myc downregulation. J Cell Sci 117 4727 4737
38. RahnamaFToftgardRZaphiropoulosPG 2004 Distinct roles of PTCH2 splice variants in Hedgehog signalling. Biochem J 378 325 334
39. MishraRGuptaSKMeiriKFFongMThostrupP 2008 GAP-43 is key to mitotic spindle control and centrosome-based polarization in neurons. Cell Cycle 7 348 357
40. ZhuYYuTZhangXCNagasawaTWuJY 2002 Role of the chemokine SDF-1 as the meningeal attractant for embryonic cerebellar neurons. Nat Neurosci 5 719 720
41. WeyerASchillingK 2003 Developmental and cell type-specific expression of the neuronal marker NeuN in the murine cerebellum. J Neurosci Res 73 400 409
42. FrappartPOLeeYRussellHRChalhoubNWangYD 2009 Recurrent genomic alterations characterize medulloblastoma arising from DNA double-strand break repair deficiency. Proc Natl Acad Sci U S A 106 1880 1885
43. BiederbickARosserRStorreJElsasserHP 2004 The VSFASSQQ motif confers calcium sensitivity to the intracellular apyrase LALP70. BMC Biochem 5 8
44. FangMShenZHuangSZhaoLChenS 2010 The ER UDPase ENTPD5 promotes protein N-glycosylation, the Warburg effect, and proliferation in the PTEN pathway. Cell 143 711 724
45. TaniguchiN 2007 A sugar-coated switch for cellular growth and arrest. Nat Chem Biol 3 307 309
46. BarnesEAKongMOllendorffVDonoghueDJ 2001 Patched1 interacts with cyclin B1 to regulate cell cycle progression. EMBO J 20 2214 2223
47. TorroglosaAMurillo-CarreteroMRomero-GrimaldiCMatarredonaERCampos-CaroA 2007 Nitric oxide decreases subventricular zone stem cell proliferation by inhibition of epidermal growth factor receptor and phosphoinositide-3-kinase/Akt pathway. Stem Cells 25 88 97
48. MatarredonaERMurillo-CarreteroMMoreno-LopezBEstradaC 2004 Nitric oxide synthesis inhibition increases proliferation of neural precursors isolated from the postnatal mouse subventricular zone. Brain Res 995 274 284
49. YuanQHuBChuTHSuHZhangW 2010 Co-expression of GAP-43 and nNOS in avulsed motoneurons and their potential role for motoneuron regeneration. Nitric Oxide 23 258 263
50. ChenTJHuangCWWangDCChenSS 2004 Co-induction of growth-associated protein GAP-43 and neuronal nitric oxide synthase in the cochlear nucleus following cochleotomy. Exp Brain Res 158 151 162
51. Lopez-JimenezMEBartolome-MartinDSanchez-PrietoJTorresM 2009 Suppression of guanylyl cyclase (beta1 subunit) expression impairs neurite outgrowth and synapse maturation in cultured cerebellar granule cells. Cell Death Differ 16 1266 1278
52. JuradoSRodriguez-PascualFSanchez-PrietoJReimundeFMLamasS 2006 NMDA induces post-transcriptional regulation of alpha2-guanylyl-cyclase-subunit expression in cerebellar granule cells. J Cell Sci 119 1622 1631
53. DeMariaCTBrewerG 1996 AUF1 binding affinity to A+U-rich elements correlates with rapid mRNA degradation. J Biol Chem 271 12179 12184
54. TsaiKCCansinoVVKohnDTNeveRLPerrone-BizzozeroNI 1997 Post-transcriptional regulation of the GAP-43 gene by specific sequences in the 3′ untranslated region of the mRNA. J Neurosci 17 1950 1958
55. ChungSEckrichMPerrone-BizzozeroNKohnDTFurneauxH 1997 The Elav-like proteins bind to a conserved regulatory element in the 3′-untranslated region of GAP-43 mRNA. J Biol Chem 272 6593 6598
56. GorgelsTGVan Lookeren CampagneMOestreicherABGribnauAAGispenWH 1989 B-50/GAP43 is localized at the cytoplasmic side of the plasma membrane in developing and adult rat pyramidal tract. J Neurosci 9 3861 3869
57. MeiriKFSaffellJLWalshFSDohertyP 1998 Neurite outgrowth stimulated by neural cell adhesion molecules requires growth-associated protein-43 (GAP-43) function and is associated with GAP-43 phosphorylation in growth cones. J Neurosci 18 10429 10437
58. ShenYManiSDonovanSLSchwobJEMeiriKF 2002 Growth-associated protein-43 is required for commissural axon guidance in the developing vertebrate nervous system. J Neurosci 22 239 247
59. ChedotalA 2010 Should I stay or should I go? Becoming a granule cell. Trends Neurosci 33 163 172
60. ZmudaJFRivasRJ 1998 The Golgi apparatus and the centrosome are localized to the sites of newly emerging axons in cerebellar granule neurons in vitro. Cell Motil Cytoskeleton 41 18 38
61. GuptaSKMeiriKFMahfoozKBhartiUManiS 2010 Coordination between extrinsic extracellular matrix cues and intrinsic responses to orient the centrosome in polarizing cerebellar granule neurons. J Neurosci 30 2755 2766
62. TegengeMABickerG 2009 Nitric oxide and cGMP signal transduction positively regulates the motility of human neuronal precursor (NT2) cells. J Neurochem 110 1828 1841
63. TanakaMYoshidaSYanoMHanaokaF 1994 Roles of endogenous nitric oxide in cerebellar cortical development in slice cultures. Neuroreport 5 2049 2052
64. ShenYMishraRManiSMeiriKF 2008 Both cell-autonomous and cell non-autonomous functions of GAP-43 are required for normal patterning of the cerebellum in vivo. Cerebellum 7 451 466
65. YavariANagarajROwusu-AnsahEFolickANgoK 2010 Role of lipid metabolism in smoothened derepression in hedgehog signaling. Dev Cell 19 54 65
66. SkwarekLCBoulianneGL 2009 Great expectations for PIP: phosphoinositides as regulators of signaling during development and disease. Dev Cell 16 12 20
67. ZakharovVVMosevitskyMI 2010 Oligomeric structure of brain abundant proteins GAP-43 and BASP1. J Struct Biol 170 470 483
68. LauxTFukamiKThelenMGolubTFreyD 2000 GAP43, MARCKS, and CAP23 modulate PI(4,5)P(2) at plasmalemmal rafts, and regulate cell cortex actin dynamics through a common mechanism. J Cell Biol 149 1455 1472
69. ArgentiBGalloRDi MarcotullioLFerrettiENapolitanoM 2005 Hedgehog antagonist REN(KCTD11) regulates proliferation and apoptosis of developing granule cell progenitors. J Neurosci 25 8338 8346
70. BlaessSGraus-PortaDBelvindrahRRadakovitsRPonsS 2004 Beta1-integrins are critical for cerebellar granule cell precursor proliferation. J Neurosci 24 3402 3412
71. GrimmerMRWeissWA 2008 BMPs oppose Math1 in cerebellar development and in medulloblastoma. Genes Dev 22 693 699
72. LaubachVESheselyEGSmithiesOShermanPA 1995 Mice lacking inducible nitric oxide synthase are not resistant to lipopolysaccharide-induced death. Proc Natl Acad Sci U S A 92 10688 10692
73. van den BoomJWolterMKuickRMisekDEYoukilisAS 2003 Characterization of gene expression profiles associated with glioma progression using oligonucleotide-based microarray analysis and real-time reverse transcription-polymerase chain reaction. Am J Pathol 163 1033 1043
74. SchlingemannJThuerigenOIttrichCToedtGKramerH 2005 Effective transcriptome amplification for expression profiling on sense-oriented oligonucleotide microarrays. Nucleic Acids Res 33 e29
75. Solinas-ToldoSLampelSStilgenbauerSNickolenkoJBennerA 1997 Matrix-based comparative genomic hybridization: biochips to screen for genomic imbalances. Genes Chromosomes Cancer 20 399 407
76. PfafflMW 2001 A new mathematical model for relative quantification in real-time RT-PCR. Nucleic Acids Res 29 e45
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Genetika Reprodukčná medicínaČlánok vyšiel v časopise
PLOS Genetics
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