Comparative multidimensional molecular analyses of pediatric diffuse intrinsic pontine glioma reveals distinct molecular subtypes
详细信息 查看全文
- 作者：Amanda M. Saratsis (1) (2)
Madhuri Kambhampati (2)
Kendall Snyder (2)
Sridevi Yadavilli (2)
Joseph M. Devaney (2) (3)
Brennan Harmon (2)
Jordan Hall (2)
Eric H. Raabe (4) (5)
Ping An (4) (6)
Melanie Weingart (4)
Brian R. Rood (7)
Suresh N. Magge (8)
Tobey J. MacDonald (9)
Roger J. Packer (10) (11)
Javad Nazarian (2) (3) - 关键词：Diffuse intrinsic pontine glioma (DIPG) ; Brainstem glioma ; Proteomics ; Histone H3 ; Myc oncogene ; Hedgehog
- 刊名：Acta Neuropathologica
- 出版年：2014
- 出版时间：June 2014
- 年：2014
- 卷：127
- 期：6
- 页码：881-895
- 全文大小：
- 参考文献：1. Ajit C, Tamhane DDD (2000) Statistics and data analysis: from elementary to intermediate. Prentice Hall, Upper Saddle River, NJ
2. Angelini P, Hawkins C, Laperriere N, Bouffet E, Bartels U (2011) Post mortem examinations in diffuse intrinsic pontine glioma: challenges and chances. J Neurooncol 101(1):75-1. doi:10.1007/s11060-010-0224-7 CrossRef
3. Barrow J, Adamowicz-Brice M, Cartmill M, MacArthur D, Lowe J, Robson K, Brundler MA, Walker DA, Coyle B, Grundy R (2011) Homozygous loss of ADAM3A revealed by genome-wide analysis of pediatric high-grade glioma and diffuse intrinsic pontine gliomas. Neuro Oncol 13(2):212-22. doi:10.1093/neuonc/noq158 CrossRef
4. Bartels U, Hawkins C, Vezina G, Kun L, Souweidane M, Bouffet E (2011) Proceedings of the diffuse intrinsic pontine glioma (DIPG) Toronto Think Tank: advancing basic and translational research and cooperation in DIPG. J Neurooncol 105(1):119-25. doi:10.1007/s11060-011-0704-4 CrossRef
5. Baudis M (2012) Progenetix oncogenomic online resource: http://www.progenetix.net. http://www.progenetix.org/cgi-bin/projectsHome.cgi?project=DIPG. Accessed May 3 2013
6. Bax DA, Mackay A, Little SE, Carvalho D, Viana-Pereira M, Tamber N, Grigoriadis AE, Ashworth A, Reis RM, Ellison DW, Al-Sarraj S, Hargrave D, Jones C (2010) A distinct spectrum of copy number aberrations in pediatric high-grade gliomas. Clin Cancer Res 16(13):3368-377. doi:10.1158/1078-0432.CCR-10-0438 CrossRef
7. Belot N, Rorive S, Doyen I, Lefranc F, Bruyneel E, Dedecker R, Micik S, Brotchi J, Decaestecker C, Salmon I, Kiss R, Camby I (2001) Molecular characterization of cell substratum attachments in human glial tumors relates to prognostic features. Glia 36(3):375-90 CrossRef
8. Broniscer A, Baker JN, Baker SJ, Chi SN, Geyer JR, Morris EB, Gajjar A (2010) Prospective collection of tissue samples at autopsy in children with diffuse intrinsic pontine glioma. Cancer 116(19):4632-637. doi:10.1002/cncr.25405 CrossRef
9. Chan KM, Fang D, Gan H, Hashizume R, Yu C, Schroeder M, Gupta N, Mueller S, James CD, Jenkins R, Sarkaria J, Zhang Z (2013) The histone H3.3K27M mutation in pediatric glioma reprograms H3K27 methylation and gene expression. Genes Dev. doi:10.1101/gad.217778.113
10. Chen D, Wang Y, Zhang K, Jiao X, Yan B, Liang J (2012) Antisense oligonucleotide against clusterin regulates human hepatocellular carcinoma invasion through transcriptional regulation of matrix metalloproteinase-2 and e-cadherin. Int J Mol Sci 13(8):10594-0607. doi:10.3390/ijms130810594 CrossRef
11. Dawson MA, Kouzarides T (2012) Cancer epigenetics: from mechanism to therapy. Cell 150(1):12-7. doi:10.1016/j.cell.2012.06.013 CrossRef
12. Dhayalan A, Tamas R, Bock I, Tattermusch A, Dimitrova E, Kudithipudi S, Ragozin S, Jeltsch A (2011) The ATRX-ADD domain binds to H3 tail peptides and reads the combined methylation state of K4 and K9. Hum Mol Genet 20(11):2195-203. doi:10.1093/hmg/ddr107 CrossRef
13. Ehrlich M, Lacey M (2013) DNA hypomethylation and hemimethylation in cancer. Adv Exp Med Biol 754:31-6. doi:10.1007/978-1-4419-9967-2_2 CrossRef
14. Eisenhart C (1947) The assumptions underlying the analysis of variance. Biometrics 1(3):1-1 CrossRef
15. Ekici AI, Eren B, Turkmen N, Comunoglu N, Fedakar R (2008) Clusterin expression in non-neoplastic adenohypophyses and pituitary adenomas: cytoplasmic clusterin localization in adenohypophysis is related to aging. Endocr Pathol 19(1):47-3. doi:10.1007/s12022-008-9015-5 CrossRef
16. Frazier JL, Lee J, Thomale UW, Noggle JC, Cohen KJ, Jallo GI (2009) Treatment of diffuse intrinsic brainstem gliomas: failed approaches and future strategies. J Neurosurg Pediatr 3(4):259-69. doi:10.3171/2008.11.PEDS08281 CrossRef
17. Fu Y, Lai Y, Wang Q, Liu X, He W, Zhang H, Fan C, Yang G (2013) Overexpression of clusterin promotes angiogenesis via the vascular endothelial growth factor in primary ovarian cancer. Mol Med Rep. doi:10.3892/mmr.2013.1436
18. Fullgrabe J, Kavanagh E, Joseph B (2011) Histone onco-modifications. Oncogene 30(31):3391-403. doi:10.1038/onc.2011.121 CrossRef
19. Gielen GH, Gessi M, Hammes J, Kramm CM, Waha A, Pietsch T (2013) H3F3A K27M mutation in pediatric CNS tumors: a marker for diffuse high-grade astrocytomas. Am J Clin Pathol 139(3):345-49. doi:10.1309/AJCPABOHBC33FVMO CrossRef
20. Gilbertson RJ, Hill DA, Hernan R, Kocak M, Geyer R, Olson J, Gajjar A, Rush L, Hamilton RL, Finkelstein SD, Pollack IF (2003) ERBB1 is amplified and overexpressed in high-grade diffusely infiltrative pediatric brain stem glioma. Clin Cancer Res 9(10 Pt 1):3620-624
21. Hankinson TC, Campagna EJ, Foreman NK, Handler MH (2011) Interpretation of magnetic resonance images in diffuse intrinsic pontine glioma: a survey of pediatric neurosurgeons. J Neurosurg Pediatr 8(1):97-02. doi:10.3171/2011.4.PEDS1180 CrossRef
22. Herranz M, Esteller M (2007) DNA methylation and histone modifications in patients with cancer: potential prognostic and therapeutic targets. Methods Mol Biol 361:25-2. doi:10.1385/1-59745-208-4:25
23. Hon GC, Hawkins RD, Caballero OL, Lo C, Lister R, Pelizzola M, Valsesia A, Ye Z, Kuan S, Edsall LE, Camargo AA, Stevenson BJ, Ecker JR, Bafna V, Strausberg RL, Simpson AJ, Ren B (2012) Global DNA hypomethylation coupled to repressive chromatin domain formation and gene silencing in breast cancer. Genome Res 22(2):246-58. doi:10.1101/gr.125872.111 CrossRef
24. Hovestadt V, Albrecht S, Kool M, Nantel A, Konermann C, Lindroth A, Jager N, Rausch T, Ryzhova M, Korbel JO, Hielscher T, Hauser P, Garami M, Klekner A, Bognar L, Ebinger M, Schuhmann MU, Scheurlen W, Pekrun A, Fruhwald MC, Roggendorf W, Kramm C, Durken M, Atkinson J, Lepage P, Montpetit A, Zakrzewska M, Zakrzewski K, Liberski PP, Dong Z, Siegel P, Kulozik AE, Zapatka M, Guha A, Malkin D, Felsberg J, Reifenberger G, von Deimling A, Ichimura K, Collins VP, Witt H, Milde T, Witt O, Zhang C, Castelo-Branco P, Lichter P, Faury D, Tabori U, Plass C, Majewski J, Pfister SM, Jabado N (2012) Driver mutations in histone H3.3 and chromatin remodelling genes in paediatric glioblastoma. Nature 482(7384):226-31. doi:10.1038/nature10833
25. Jansen MH, van Vuurden DG, Vandertop WP, Kaspers GJ (2012) Diffuse intrinsic pontine gliomas: a systematic update on clinical trials and biology. Cancer Treat Rev 38(1):27-5. doi:10.1016/j.ctrv.2011.06.007 CrossRef
26. Khan AA, Dace DS, Ryazanov AG, Kelly J, Apte RS (2010) Resveratrol regulates pathologic angiogenesis by a eukaryotic elongation factor-2 kinase-regulated pathway. Am J Pathol 177(1):481-92. doi:10.2353/ajpath.2010.090836 CrossRef
27. Khuong-Quang DA, Buczkowicz P, Rakopoulos P, Liu XY, Fontebasso AM, Bouffet E, Bartels U, Albrecht S, Schwartzentruber J, Letourneau L, Bourgey M, Bourque G, Montpetit A, Bourret G, Lepage P, Fleming A, Lichter P, Kool M, von Deimling A, Sturm D, Korshunov A, Faury D, Jones DT, Majewski J, Pfister SM, Jabado N, Hawkins C (2012) K27M mutation in histone H3.3 defines clinically and biologically distinct subgroups of pediatric diffuse intrinsic pontine gliomas. Acta Neuropathol 124(3):439-47. doi:10.1007/s00401-012-0998-0 CrossRef
28. Khuong-Quang DA, Gerges N, Jabado N (2012) Mutations in histone H3.3 and chromatin remodeling genes drive pediatric and young adult glioblastomas. Med Sci (Paris) 28(10):809-12. doi:10.1051/medsci/20122810004 CrossRef
29. Kool M, Korshunov A, Remke M, Jones DT, Schlanstein M, Northcott PA, Cho YJ, Koster J, Schouten-van Meeteren A, van Vuurden D, Clifford SC, Pietsch T, von Bueren AO, Rutkowski S, McCabe M, Collins VP, Backlund ML, Haberler C, Bourdeaut F, Delattre O, Doz F, Ellison DW, Gilbertson RJ, Pomeroy SL, Taylor MD, Lichter P, Pfister SM (2012) Molecular subgroups of medulloblastoma: an international meta-analysis of transcriptome, genetic aberrations, and clinical data of WNT, SHH, Group 3, and Group 4 medulloblastomas. Acta Neuropathol 123(4):473-84. doi:10.1007/s00401-012-0958-8 CrossRef
30. Lewis PW, Elsaesser SJ, Noh KM, Stadler SC, Allis CD (2010) Daxx is an H3.3-specific histone chaperone and cooperates with ATRX in replication-independent chromatin assembly at telomeres. Proc Natl Acad Sci U S A 107(32):14075-4080. doi:10.1073/pnas.1008850107 CrossRef
31. Lewis PW, Muller MM, Koletsky MS, Cordero F, Lin S, Banaszynski LA, Garcia BA, Muir TW, Becher OJ, Allis CD (2013) Inhibition of PRC2 activity by a gain-of-function H3 mutation found in pediatric glioblastoma. Science. doi:10.1126/science.1232245
32. Ling IF, Bhongsatiern J, Simpson JF, Fardo DW, Estus S (2012) Genetics of clusterin isoform expression and Alzheimer’s disease risk. PLoS ONE 7(4):e33923. doi:10.1371/journal.pone.0033923 CrossRef
33. Mace EM, Zhang J, Siminovitch KA, Takei F (2010) Elucidation of the integrin LFA-1-mediated signaling pathway of actin polarization in natural killer cells. Blood 116(8):1272-279. doi:10.1182/blood-2009-12-261487 CrossRef
34. Monje M, Mitra SS, Freret ME, Raveh TB, Kim J, Masek M, Attema JL, Li G, Haddix T, Edwards MS, Fisher PG, Weissman IL, Rowitch DH, Vogel H, Wong AJ, Beachy PA (2011) Hedgehog-responsive candidate cell of origin for diffuse intrinsic pontine glioma. Proc Natl Acad Sci USA 108(11):4453-458. doi:10.1073/pnas.1101657108 CrossRef
35. Nazarian J, Santi M, Hathout Y, Macdonald TJ (2008) Protein profiling of formalin fixed paraffin embedded tissue: identification of potential biomarkers for pediatric brainstem glioma. Proteomics Clin Appl 2(6):915-24. doi:10.1002/prca.200780061 CrossRef
36. Northcott PA, Korshunov A, Witt H, Hielscher T, Eberhart CG, Mack S, Bouffet E, Clifford SC, Hawkins CE, French P, Rutka JT, Pfister S, Taylor MD (2011) Medulloblastoma comprises four distinct molecular variants. J Clin Oncol 29(11):1408-414. doi:10.1200/JCO.2009.27.4324 CrossRef
37. Paugh BS, Qu C, Jones C, Liu Z, Adamowicz-Brice M, Zhang J, Bax DA, Coyle B, Barrow J, Hargrave D, Lowe J, Gajjar A, Zhao W, Broniscer A, Ellison DW, Grundy RG, Baker SJ (2010) Integrated molecular genetic profiling of pediatric high-grade gliomas reveals key differences with the adult disease. J Clin Oncol 28(18):3061-068. doi:10.1200/JCO.2009.26.7252 CrossRef
38. Perez-Gomez JL, Rodriguez-Alvarez CA, Marhx-Bracho A, Rueda-Franco F (2010) Stereotactic biopsy for brainstem tumors in pediatric patients. Childs Nerv Syst 26(1):29-4. doi:10.1007/s00381-009-1000-x CrossRef
39. Puget S, Philippe C, Bax DA, Job B, Varlet P, Junier MP, Andreiuolo F, Carvalho D, Reis R, Guerrini-Rousseau L, Roujeau T, Dessen P, Richon C, Lazar V, Le Teuff G, Sainte-Rose C, Geoerger B, Vassal G, Jones C, Grill J (2012) Mesenchymal transition and PDGFRA amplification/mutation are key distinct oncogenic events in pediatric diffuse intrinsic pontine gliomas. PLoS ONE 7(2):e30313. doi:10.1371/journal.pone.0030313 CrossRef
40. Qureshi AH, Chaoji V, Maiguel D, Faridi MH, Barth CJ, Salem SM, Singhal M, Stoub D, Krastins B, Ogihara M, Zaki MJ, Gupta V (2009) Proteomic and phospho-proteomic profile of human platelets in basal, resting state: insights into integrin signaling. PLoS ONE 4(10):e7627. doi:10.1371/journal.pone.0007627 CrossRef
41. Rao Malla R, Gopinath S, Alapati K, Gorantla B, Gondi CS, Rao JS (2012) Knockdown of cathepsin B and uPAR inhibits CD151 and alpha3beta1 integrin-mediated cell adhesion and invasion in glioma. Mol Carcinog. doi:10.1002/mc.21915
42. Ratnikov BI, Partridge AW, Ginsberg MH (2005) Integrin activation by talin. J Thromb Haemost 3(8):1783-790. doi:10.1111/j.1538-7836.2005.01362.x CrossRef
43. Redondo M, Villar E, Torres-Munoz J, Tellez T, Morell M, Petito CK (2000) Overexpression of clusterin in human breast carcinoma. Am J Pathol 157(2):393-99. doi:10.1016/S0002-9440(10)64552-X CrossRef
44. Saratsis AM, Yadavilli S, Magge S, Rood BR, Perez J, Hill DA, Hwang E, Kilburn L, Packer RJ, Nazarian J (2012) Insights into pediatric diffuse intrinsic pontine glioma through proteomic analysis of cerebrospinal fluid. Neuro Oncol 14(5):547-60. doi:10.1093/neuonc/nos067 CrossRef
45. Scaringi C, Minniti G, Caporello P, Enrici RM (2012) Integrin inhibitor cilengitide for the treatment of glioblastoma: a brief overview of current clinical results. Anticancer Res 32(10):4213-223
46. Sen S, Ng WP, Kumar S (2012) Contributions of talin-1 to glioma cell-matrix tensional homeostasis. J R Soc Interface 9(71):1311-317. doi:10.1098/rsif 2011.0567 CrossRef
47. Sharma S, Free A, Mei Y, Peiper SC, Wang Z, Cowell JK (2010) Distinct molecular signatures in pediatric infratentorial glioblastomas defined by aCGH. Exp Mol Pathol 89(2):169-74. doi:10.1016/j.yexmp.2010.06.009 CrossRef
48. Sturm D, Witt H, Hovestadt V, Khuong-Quang DA, Jones DT, Konermann C, Pfaff E, Tonjes M, Sill M, Bender S, Kool M, Zapatka M, Becker N, Zucknick M, Hielscher T, Liu XY, Fontebasso AM, Ryzhova M, Albrecht S, Jacob K, Wolter M, Ebinger M, Schuhmann MU, van Meter T, Fruhwald MC, Hauch H, Pekrun A, Radlwimmer B, Niehues T, von Komorowski G, Durken M, Kulozik AE, Madden J, Donson A, Foreman NK, Drissi R, Fouladi M, Scheurlen W, von Deimling A, Monoranu C, Roggendorf W, Herold-Mende C, Unterberg A, Kramm CM, Felsberg J, Hartmann C, Wiestler B, Wick W, Milde T, Witt O, Lindroth AM, Schwartzentruber J, Faury D, Fleming A, Zakrzewska M, Liberski PP, Zakrzewski K, Hauser P, Garami M, Klekner A, Bognar L, Morrissy S, Cavalli F, Taylor MD, van Sluis P, Koster J, Versteeg R, Volckmann R, Mikkelsen T, Aldape K, Reifenberger G, Collins VP, Majewski J, Korshunov A, Lichter P, Plass C, Jabado N, Pfister SM (2012) Hotspot mutations in H3F3A and IDH1 define distinct epigenetic and biological subgroups of glioblastoma. Cancer Cell 22(4):425-37. doi:10.1016/j.ccr.2012.08.024 CrossRef
49. Sufit A, Donson AM, Birks DK, Knipstein JA, Fenton LZ, Jedlicka P, Hankinson TC, Handler MH, Foreman NK (2012) Diffuse intrinsic pontine tumors: a study of primitive neuroectodermal tumors versus the more common diffuse intrinsic pontine gliomas. J Neurosurg Pediatr 10(2):81-8. doi:10.3171/2012.3.PEDS11316 CrossRef
50. Tabatabai G, Weller M, Nabors B, Picard M, Reardon D, Mikkelsen T, Ruegg C, Stupp R (2010) Targeting integrins in malignant glioma. Target Oncol 5(3):175-81. doi:10.1007/s11523-010-0156-3 CrossRef
51. Taylor MD, Northcott PA, Korshunov A, Remke M, Cho YJ, Clifford SC, Eberhart CG, Parsons DW, Rutkowski S, Gajjar A, Ellison DW, Lichter P, Gilbertson RJ, Pomeroy SL, Kool M, Pfister SM (2012) Molecular subgroups of medulloblastoma: the current consensus. Acta Neuropathol 123(4):465-72. doi:10.1007/s00401-011-0922-z CrossRef
52. Venneti S, Garimella MT, Sullivan LM, Martinez D, Huse JT, Heguy A, Santi M, Thompson CB, Judkins AR (2013) Evaluation of Histone 3 Lysine 27 Trimethylation (H3K27me3) and Enhancer of Zest 2 (EZH2) in pediatric glial and glioneuronal tumors shows decreased H3K27me3 in H3F3A K27M mutant glioblastomas. Brain Pathol. doi:10.1111/bpa.12042
53. Walker DA, Punt JA, Sokal M (1999) Clinical management of brain stem glioma. Arch Dis Child 80(6):558-64 CrossRef
54. Wang C, Jiang K, Kang X, Gao D, Sun C, Li Y, Sun L, Zhang S, Liu X, Wu W, Yang P, Guo K, Liu Y (2012) Tumor-derived secretory clusterin induces epithelial–mesenchymal transition and facilitates hepatocellular carcinoma metastasis. Int J Biochem Cell Biol 44(12):2308-320. doi:10.1016/j.biocel.2012.09.012 CrossRef
55. Wang Y, Wang X, Zhao H, Liang B, Du Q (2012) Clusterin confers resistance to TNF-alpha-induced apoptosis in breast cancer cells through NF-kappaB activation and Bcl-2 overexpression. J Chemother 24(6):348-57. doi:10.1179/1973947812Y.0000000049 CrossRef
56. Wegener KL, Partridge AW, Han J, Pickford AR, Liddington RC, Ginsberg MH, Campbell ID (2007) Structural basis of integrin activation by talin. Cell 128(1):171-82. doi:10.1016/j.cell.2006.10.048 CrossRef
57. Wu G, Broniscer A, McEachron TA, Lu C, Paugh BS, Becksfort J, Qu C, Ding L, Huether R, Parker M, Zhang J, Gajjar A, Dyer MA, Mullighan CG, Gilbertson RJ, Mardis ER, Wilson RK, Downing JR, Ellison DW, Baker SJ (2012) Somatic histone H3 alterations in pediatric diffuse intrinsic pontine gliomas and non-brainstem glioblastomas. Nat Genet 44(3):251-53. doi:10.1038/ng.1102 CrossRef
58. Zarghooni M, Bartels U, Lee E, Buczkowicz P, Morrison A, Huang A, Bouffet E, Hawkins C (2010) Whole-genome profiling of pediatric diffuse intrinsic pontine gliomas highlights platelet-derived growth factor receptor alpha and poly (ADP-ribose) polymerase as potential therapeutic targets. J Clin Oncol 28(8):1337-344. doi:10.1200/JCO.2009.25.5463 CrossRef - 作者单位：Amanda M. Saratsis (1) (2)
Madhuri Kambhampati (2)
Kendall Snyder (2)
Sridevi Yadavilli (2)
Joseph M. Devaney (2) (3)
Brennan Harmon (2)
Jordan Hall (2)
Eric H. Raabe (4) (5)
Ping An (4) (6)
Melanie Weingart (4)
Brian R. Rood (7)
Suresh N. Magge (8)
Tobey J. MacDonald (9)
Roger J. Packer (10) (11)
Javad Nazarian (2) (3)
1. Division of Neurosurgery, Ann & Robert H. Lurie Children’s Hospital of Chicago, 225 E. Chicago Avenue, Chicago, IL, 60611, USA
2. Center for Genetic Medicine Research, Children’s National Medical Center, 111 Michigan Ave., NW, Washington, DC, 20010, USA
3. Department of Integrative Systems Biology, George Washington University School of Medicine and Health Sciences, Washington, DC, 20052, USA
4. Division of Neuro-Pathology, Johns Hopkins University School of Medicine, Baltimore, MD, 21287, USA
5. Division of Pediatric Oncology, Johns Hopkins University School of Medicine, Baltimore, MD, 21287, USA
6. Neurobiology Department, College of Basic Medical Sciences, China Medical University, Shenyang, 110001, China
7. Division of Oncology, Center for Cancer and Immunology Research, Children’s National Medical Center, Washington, DC, 20010, USA
8. Division of Neurosurgery, Children’s National Medical Center, Washington, DC, 20010, USA
9. Department of Pediatrics, Emory University School of Medicine, Atlanta, GA, 30322, USA
10. Division of Neurology, Center for Neuroscience Research, Children’s National Medical Center, Washington, DC, 20010, USA
11. Brain Tumor Institute, Center for Neuroscience and Behavioral Medicine, Children’s National Medical Center, Washington, DC, USA - ISSN：1432-0533
文摘
Diffuse intrinsic pontine glioma (DIPG) is a highly morbid form of pediatric brainstem glioma. Here, we present the first comprehensive protein, mRNA, and methylation profiles of fresh-frozen DIPG specimens (n?=?14), normal brain tissue (n?=?10), and other pediatric brain tumors (n?=?17). Protein profiling identified 2,305 unique proteins indicating distinct DIPG protein expression patterns compared to other pediatric brain tumors. Western blot and immunohistochemistry validated upregulation of Clusterin (CLU), Elongation Factor 2 (EF2), and Talin-1 (TLN1) in DIPGs studied. Comparisons to mRNA expression profiles generated from tumor and adjacent normal brain tissue indicated two DIPG subgroups, characterized by upregulation of Myc (N-Myc) or Hedgehog (Hh) signaling. We validated upregulation of PTCH, a membrane receptor in the Hh signaling pathway, in a subgroup of DIPG specimens. DNA methylation analysis indicated global hypomethylation of DIPG compared to adjacent normal tissue specimens, with differential methylation of 24 genes involved in Hh and Myc pathways, correlating with protein and mRNA expression patterns. Sequencing analysis showed c.83A>T mutations in the H3F3A or HIST1H3B gene in 77?% of our DIPG cohort. Supervised analysis revealed a unique methylation pattern in mutated specimens compared to the wild-type DIPG samples. This study presents the first comprehensive multidimensional protein, mRNA, and methylation profiling of pediatric brain tumor specimens, detecting the presence of two subgroups within our DIPG cohort. This multidimensional analysis of DIPG provides increased analytical power to more fully explore molecular signatures of DIPGs, with implications for evaluating potential molecular subtypes and biomarker discovery for assessing response to therapy.