title: Overexpression of mutant Ptch in rhabdomyosarcomas is associated with promoter hypomethylation and increased Gli1 and H3K4me3 occupancy.
abstract: Mice with heterozygous loss of the tumor suppressor Patched1 (Ptch) develop rhabdomyosarcoma (RMS)-like tumors. However, Ptch transcripts are consistently overexpressed in these tumors. We have recently shown that the upregulated transcripts are derived from the mutated Ptch allele thus leading to the hypothesis that the wild-type allele is repressed during RMS development. Here we describe epigenetic changes taking place at the Ptch locus during RMS development. We showed a lower degree of DNA-methylation in methylation-sensitive CpG regions of the Ptch promoter in RMS compared to normal muscle from heterozygous Ptch animals. In agreement with these results, treatment of heterozygous Ptch mice with the DNA demethylating agent 5-aza-2-deoxycytidine (5-aza-dC) between embryonic days E9.5-E11.5 significantly accelerated RMS formation. Since Ptch promoter methylation occurs after/around E13.5, the window for RMS initiation during embryogenesis, these results provide additional evidence that Ptch promoter hypomethylation may contribute to RMS formation. We have also demonstrated increased trimethylation of histone H3 lysine 4 (H3K4me3) and preferential binding of Gli1, a known Ptch activator, to the mutant locus in RMS. Together, these findings support an alternative model for RMS formation in heterozygous Ptch mice including loss of methylation and concomitant occupancy by activating histone marks of mutant Ptch.
authors.affiliation: Department of Human Genetics, University Medical Center, Göttingen, Germany.
authors.forename: Frauke
authors.initials: F
authors.lastname: Nitzki
authors.affiliation: Schulze Center for Novel Therapeutics, Division of Oncology Research, Mayo Clinic, Rochester, MN, USA.
authors.forename: Ezequiel J
authors.initials: EJ
authors.lastname: Tolosa
authors.affiliation: Department of Human Genetics, University Medical Center, Göttingen, Germany.
authors.forename: Nicole
authors.initials: N
authors.lastname: Cuvelier
authors.affiliation: Department of Human Genetics, University Medical Center, Göttingen, Germany.
authors.forename: Anke
authors.initials: A
authors.lastname: Frommhold
authors.affiliation: Microarray and Deep-Sequencing Core Facility, University Medical Center, Göttingen, Germany.
authors.forename: Gabriela
authors.initials: G
authors.lastname: Salinas-Riester
authors.affiliation: Department of General, Visceral and Pediatric Surgery, University Medical Center Göttingen, Göttingen, Germany.
authors.forename: Steven A
authors.initials: SA
authors.lastname: Johnsen
authors.affiliation: Schulze Center for Novel Therapeutics, Division of Oncology Research, Mayo Clinic, Rochester, MN, USA.
authors.forename: Martin E
authors.initials: ME
authors.lastname: Fernandez-Zapico
authors.affiliation: Department of Human Genetics, University Medical Center, Göttingen, Germany.
authors.forename: Heidi
authors.initials: H
authors.lastname: Hahn
mesh_terms: D000818:Animals; D001374:Azacitidine / Q000031:analogs & derivatives / Q000494:pharmacology; D018899:CpG Islands; D019175:DNA Methylation / Q000187:drug effects; D000077209:Decitabine; D047108:Embryonic Development / Q000235:genetics; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects / Q000235:genetics; D016147:Genes, Tumor Suppressor; D005865:Gestational Age; D006579:Heterozygote; D006657:Histones / Q000378:metabolism*; D051741:Kruppel-Like Transcription Factors / Q000378:metabolism*; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D008957:Models, Genetic; D009154:Mutation; D009363:Neoplasm Proteins / Q000096:biosynthesis / Q000235:genetics / Q000502:physiology*; D000072079:Patched Receptors; D000072081:Patched-1 Receptor; D020641:Polymorphism, Single Nucleotide; D011401:Promoter Regions, Genetic / Q000235:genetics*; D011499:Protein Processing, Post-Translational; D011956:Receptors, Cell Surface / Q000096:biosynthesis / Q000235:genetics / Q000502:physiology*; D012208:Rhabdomyosarcoma / Q000235:genetics* / Q000378:metabolism; D015398:Signal Transduction; D000071676:Zinc Finger Protein GLI1
chemical_list: C498520:Gli1 protein, mouse; D006657:Histones; D051741:Kruppel-Like Transcription Factors; D009363:Neoplasm Proteins; C000606236:PTCH1 protein, human; D000072079:Patched Receptors; D000072081:Patched-1 Receptor; C000606240:Ptch1 protein, mouse; D011956:Receptors, Cell Surface; D000071676:Zinc Finger Protein GLI1; D000077209:Decitabine; D001374:Azacitidine
keywords: DNA hypomethylation; Gli1; H3K4me3; Ptch; RMS
references.citation: Genes Dev. 2004 Nov 1;18(21):2627-38
references.pmid: 15520282
references.citation: Blood. 2007 Sep 15;110(6):1814-23
references.pmid: 17536012
references.citation: Int J Oncol. 2011 Mar;38(3):685-92
references.pmid: 21206974
references.citation: Nat Genet. 2009 Jan;41(1):125-9
references.pmid: 19098913
references.citation: Nat Rev Mol Cell Biol. 2005 Apr;6(4):306-17
references.pmid: 15803137
references.citation: Nat Cell Biol. 2010 Feb;12(2):132-42
references.pmid: 20081843
references.citation: Int J Mol Med. 2007 Jun;19(6):875-83
references.pmid: 17487419
references.citation: PLoS One. 2011;6(11):e27684
references.pmid: 22110720
references.citation: Nat Rev Cancer. 2008 Oct;8(10):743-54
references.pmid: 18813320
references.citation: Carcinogenesis. 2002 May;23(5):727-33
references.pmid: 12016144
references.citation: Proc Natl Acad Sci U S A. 2010 Jan 5;107(1):234-9
references.pmid: 19966297
references.citation: Cell. 1999 Oct 29;99(3):247-57
references.pmid: 10555141
references.citation: Cancer Genet Cytogenet. 2008 Jan 1;180(1):47-50
references.pmid: 18068533
references.citation: J Pathol. 2006 Jan;208(1):17-25
references.pmid: 16294371
references.citation: Mol Cell Biol. 2007 Dec;27(24):8466-79
references.pmid: 17923682
references.citation: Gene. 2004 Apr 14;330:101-14
references.pmid: 15087129
references.citation: Teratology. 1994 Nov;50(5):332-9
references.pmid: 7536356
references.citation: Clin Epigenetics. 2012 Mar 12;4(1):5
references.pmid: 22414492
references.citation: Biochimie. 2012 Nov;94(11):2202-11
references.pmid: 22634371
references.citation: BMC Cancer. 2010;10:614
references.pmid: 21059263
references.citation: Development. 2005 May;132(10):2425-39
references.pmid: 15843415
references.citation: Nature. 2010 Jul 8;466(7303):253-7
references.pmid: 20613842
references.citation: Nat Genet. 1996 Sep;14(1):78-81
references.pmid: 8782823
references.citation: EMBO Rep. 2011 Jul;12(7):647-56
references.pmid: 21660058
references.citation: Cancer Res. 2009 Feb 1;69(3):887-95
references.pmid: 19155313
references.citation: Cancer. 2011 Feb 15;117(4):677-87
references.pmid: 20945317
references.citation: Cancer Res. 1997 Mar 1;57(5):842-5
references.pmid: 9041183
references.citation: Toxicology. 1996 Aug 1;112(1):37-43
references.pmid: 8792847
references.citation: Clin Cancer Res. 2006 Oct 15;12(20 Pt 1):5924-8
references.pmid: 17062662
references.citation: Nat Genet. 2007 Mar;39(3):289-90
references.pmid: 17325677
references.citation: Int J Oncol. 2005 Dec;27(6):1567-75
references.pmid: 16273213
references.citation: Genomics. 2004 Nov;84(5):853-8
references.pmid: 15475264
references.citation: Cancer Res. 2014 Jul 1;74(13):3617-29
references.pmid: 24786786
references.citation: Oncogene. 2011 Oct 27;30(43):4428-36
references.pmid: 21602886
references.citation: Mol Cell. 2007 Jan 12;25(1):15-30
references.pmid: 17218268
references.citation: Genomics. 2005 Apr;85(4):462-71
references.pmid: 15780749
references.citation: Cancer Res. 1997 Jun 15;57(12):2369-72
references.pmid: 9192811
references.citation: Nat Genet. 2003 Jan;33(1):61-5
references.pmid: 12496760
references.citation: Nat Med. 1998 May;4(5):619-22
references.pmid: 9585239
references.citation: Exp Dermatol. 2010 Oct;19(10):926-8
references.pmid: 20849535
references.citation: Carcinogenesis. 2009 Jun;30(6):918-26
references.pmid: 19321799
references.citation: Nucleic Acids Res. 2006;34(1):e2
references.pmid: 16397291
references.citation: Am J Hum Genet. 2010 Feb 12;86(2):196-212
references.pmid: 20159110
references.citation: World J Gastroenterol. 2009 Aug 14;15(30):3799-806
references.pmid: 19673023
references.citation: Genome Med. 2012 May 28;4(5):47
references.pmid: 22640407
references.citation: Mol Cancer Res. 2004 Jan;2(1):62-72
references.pmid: 14757847
references.citation: Genes Dev. 2012 Aug 1;26(15):1714-28
references.pmid: 22855832