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Molecular Cancer
Published in: Molecular Cancer 1/2010

Open Access 01-12-2010 | Research

Silencing of TESTIN by dense biallelic promoter methylation is the most common molecular event in childhood acute lymphoblastic leukaemia

Authors: Robert J. Weeks, Ursula R. Kees, Sarah Song, Ian M. Morison

Published in: Molecular Cancer | Issue 1/2010

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Abstract

Background

Aberrant promoter DNA methylation has been reported in childhood acute lymphoblastic leukaemia (ALL) and has the potential to contribute to its onset and outcome. However, few reports demonstrate consistent, prevalent and dense promoter methylation, associated with tumour-specific gene silencing. By screening candidate genes, we have detected frequent and dense methylation of the TESTIN (TES) promoter.

Results

Bisulfite sequencing showed that 100% of the ALL samples (n = 20) were methylated at the TES promoter, whereas the matched remission (n = 5), normal bone marrow (n = 6) and normal PBL (n = 5) samples were unmethylated. Expression of TES in hyperdiploid, TEL-AML+, BCR-ABL+, and E2A-PBX+ subtypes of B lineage ALL was markedly reduced compared to that in normal bone marrow progenitor cells and in B cells. In addition TES methylation and silencing was demonstrated in nine out of ten independent B ALL propagated as xenografts in NOD/SCID mice.

Conclusion

In total, 93% of B ALL samples (93 of 100) demonstrated methylation with silencing or reduced expression of the TES gene. Thus, TES is the most frequently methylated and silenced gene yet reported in ALL. TES, a LIM domain-containing tumour suppressor gene and component of the focal adhesion complex, is involved in adhesion, motility, cell-to-cell interactions and cell signalling. Our data implicate TES methylation in ALL and provide additional evidence for the involvement of LIM domain proteins in leukaemogenesis.
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Literature
1.
Grady WM, Willis J, Guilford PJ, Dunbier AK, Toro TT, Lynch H, Wiesner G, Ferguson K, Eng C, Park JG, Kim SJ, Markowitz S: Methylation of the CDH1 promoter as the second genetic hit in hereditary diffuse gastric cancer. Nat Genet. 2000, 26: 16-17. 10.1038/79120CrossRefPubMed
2.
Stirzaker C, Millar DS, Paul CL, Warnecke PM, Harrison J, Vincent PC, Frommer M, Clark SJ: Extensive DNA methylation spanning the Rb promoter in retinoblastoma tumors. Cancer Res. 1997, 57: 2229-2237.PubMed
3.
Clifford SC, Prowse AH, Affara NA, Buys CH, Maher ER: Inactivation of the von Hippel-Lindau (VHL) tumour suppressor gene and allelic losses at chromosome arm 3p in primary renal cell carcinoma: evidence for a VHL-independent pathway in clear cell renal tumourigenesis. Genes Chromosomes Cancer. 1998, 22: 200-209. 10.1002/(SICI)1098-2264(199807)22:3<200::AID-GCC5>3.0.CO;2-#CrossRefPubMed
4.
Keshet I, Schlesinger Y, Farkash S, Rand E, Hecht M, Segal E, Pikarski E, Young R, Niveleau A, Cedar H, Simon I: Evidence for an instructive mechanism of de novo methylation in cancer cells. Nat Genet. 2006, 38: 149-153. 10.1038/ng1719CrossRefPubMed
5.
Paulsson K, An Q, Moorman AV, Parker H, Molloy G, Davies T, Griffiths M, Ross FM, Irving J, Harrison CJ, Young B, Strefford J: Methylation of tumour suppressor gene promoters in the presence and absence of transcriptional silencing in high hyperdiploid acute lymphoblastic leukaemia. Br J Haematol. 2009, 144: 838-847. 10.1111/j.1365-2141.2008.07523.xCrossRefPubMed
6.
Roman-Gomez J, Jimenez-Velasco A, Barrios M, Prosper F, Heiniger A, Torres A, Agirre X: Poor prognosis in acute lymphoblastic leukemia may relate to promoter hypermethylation of cancer-related genes. Leukemia & Lymphoma. 2007, 48: 1269-1282. 10.1080/10428190701344899CrossRef
7.
Taylor KH, Pena-Hernandez KE, Davis JW, Arthur GL, Duff DJ, Shi H, Rahmatpanah FB, Sjahputera O, Caldwell CW: Large-scale CpG methylation analysis identifies novel candidate genes and reveals methylation hotspots in acute lymphoblastic leukemia. Cancer Research. 2007, 67: 2617-2625. 10.1158/0008-5472.CAN-06-3993CrossRefPubMed
8.
Garcia-Manero G, Yang H, Kuang SQ, O'Brien S, Thomas D, Kantarjian H: Epigenetics of acute lymphocytic leukemia. Semin Hematol. 2009, 46: 24-32. 10.1053/j.seminhematol.2008.09.008CrossRefPubMed
9.
Davidsson J, Lilljebjorn H, Andersson A, Veerla S, Heldrup J, Behrendtz M, Fioretos T, Johansson B: The DNA methylome of pediatric acute lymphoblastic leukemia. Hum Mol Genet. 2009, 18 (21): 4054-65. 10.1093/hmg/ddp354CrossRefPubMed
10.
Chambon-Pautas C, Cave H, Gerard B, Guidal-Giroux C, Duval M, Vilmer E, Grandchamp B: High-resolution allelotype analysis of childhood B-lineage acute lymphoblastic leukemia. Leukemia. 1998, 12: 1107-1113. 10.1038/sj.leu.2401069CrossRefPubMed
11.
Takeuchi S, Koike M, Seriu T, Bartram CR, Slater J, Park S, Miyoshi I, Koeffler HP: Homozygous deletions at 9p21 in childhood acute lymphoblastic leukemia detected by microsatellite analysis. Leukemia. 1997, 11: 1636-1640. 10.1038/sj.leu.2400817CrossRefPubMed
12.
Sakashita K, Koike K, Kinoshita T, Shiohara M, Kamijo T, Taniguchi S, Kubota T: Dynamic DNA methylation change in the CpG island region of p15 during human myeloid development. J Clin Invest. 2001, 108: 1195-1204.PubMedCentralCrossRefPubMed
13.
Mirebeau D, Acquaviva C, Suciu S, Bertin R, Dastugue N, Robert A, Boutard P, Mechinaud F, Plouvier E, Otten J, Vilmer E, Cavé H, : The prognostic significance of CDKN2A, CDKN2B and MTAP inactivation in B-lineage acute lymphoblastic leukemia of childhood. Results of the EORTC studies 58881 and 58951. Haematologica. 2006, 91: 881-885.PubMed
14.
Roman-Gomez J, Jimenez-Velasco A, Castillejo JA, Agirre X, Barrios M, Navarro G, Molina FJ, Calasanz MJ, Prosper F, Heiniger A, Torres A: Promoter hypermethylation of cancer-related genes: a strong independent prognostic factor in acute lymphoblastic leukemia. Blood. 2004, 104: 2492-2498. 10.1182/blood-2004-03-0954CrossRefPubMed
15.
Tsellou E, Troungos C, Moschovi M, Athanasiadou-Piperopoulou F, Polychronopoulou S, Kosmidis H, Kalmanti M, Hatzakis A, Dessypris N, Kalofoutis A, Petridou E: Hypermethylation of CpG islands in the promoter region of the p15INK4B gene in childhood acute leukaemia. Eur J Cancer. 2005, 41: 584-589. 10.1016/j.ejca.2004.12.010CrossRefPubMed
16.
Yang Y, Takeuchi S, Hofmann WK, Ikezoe T, van Dongen JJ, Szczepanski T, Bartram CR, Yoshino N, Taguchi H, Koeffler HP: Aberrant methylation in promoter-associated CpG islands of multiple genes in acute lymphoblastic leukemia. Leuk Res. 2006, 30: 98-102. 10.1016/j.leukres.2005.06.002CrossRefPubMed
17.
Nygren AO, Ameziane N, Duarte HM, Vijzelaar RN, Waisfisz Q, Hess CJ, Schouten JP, Errami A: Methylation-specific MLPA (MS-MLPA): simultaneous detection of CpG methylation and copy number changes of up to 40 sequences. Nucleic Acids Research. 2005, 33: e128- 10.1093/nar/gni127PubMedCentralCrossRefPubMed
18.
Tatarelli C, Linnenbach A, Mimori K, Croce CM: Characterization of the human TESTIN gene localized in the FRA7G region at 7q31.2. Genomics. 2000, 68: 1-12. 10.1006/geno.2000.6272CrossRefPubMed
19.
Tobias ES, Hurlstone AF, MacKenzie E, McFarlane R, Black DM: The TES gene at 7q31.1 is methylated in tumours and encodes a novel growth-suppressing LIM domain protein. Oncogene. 2001, 20: 2844-2853. 10.1038/sj.onc.1204433CrossRefPubMed
20.
Mueller W, Nutt C, Ehrich M, Riemenschneider M, Von Deimling A, Van Den Boom D, Louis D: Downregulation of RUNX3 and TES by hypermethylation in glioblastoma. Oncogene. 2007, 26: 583-593. 10.1038/sj.onc.1209805CrossRefPubMed
21.
Yeoh EJ, Ross ME, Shurtleff S, Williams WK, Patel D, Mahfouz R, Behm FG, Raimondi SC, Relling M, Patel A, Cheng C, Campana D, Wilkins D, Zhou X, Li J, Liu H, Pui CH, Evans WE, Naeve C, Wong L, Downing JR: Classification, subtype discovery, and prediction of outcome in pediatric acute lymphoblastic leukemia by gene expression profiling. Cancer Cell. 2002, 1: 133-143. 10.1016/S1535-6108(02)00032-6CrossRefPubMed
22.
Boag J, Beesley A, Firth M, Freitas J, Ford J, Brigstock D, De Klerk N, Kees U: High expression of connective tissue growth factor in pre-B acute lymphoblastic leukaemia. Br J Haematol. 2007, 138: 740-748. 10.1111/j.1365-2141.2007.06739.xCrossRefPubMed
23.
Liem NL, Papa RA, Milross CG, Schmid MA, Tajbakhsh M, Choi S, Ramirez CD, Rice AM, Haber M, Norris MD, MacKenzie KL, Lock RB: Characterization of childhood acute lymphoblastic leukemia xenograft models for the preclinical evaluation of new therapies. Blood. 2004, 103: 3905-3914. 10.1182/blood-2003-08-2911CrossRefPubMed
24.
Hesson LB, Dunwell TL, Cooper WN, Catchpoole D, Brini AT, Chiaramonte R, Griffiths M, Chalmers AD, Maher ER, Latif F: The novel RASSF6 and RASSF10 candidate tumour suppressor genes are frequently epigenetically inactivated in childhood leukaemias. Mol Cancer. 2009, 8: 42- 10.1186/1476-4598-8-42PubMedCentralCrossRefPubMed
25.
Cigognini D, Corneo G, Fermo E, Zanella A, Tripputi P: HIC gene, a candidate suppressor gene within a minimal region of loss at 7q31.1 in myeloid neoplasms. Leuk Res. 2007, 31: 477-482. 10.1016/j.leukres.2006.09.007CrossRefPubMed
26.
Xia JC, Weng DS, Li JT, Qin HD, Mai SJ, Feng BJ, Fan Q, Feng QS, Huang LX, Yu XJ, Pan ZZ, Li YQ, Wang QJ, Zhan YQ, Chen SP, He J, Huang WL, Wu PH, Zeng YX: Loss of heterozygosity analysis of a candidate gastric carcinoma tumor suppressor locus at 7q31. Cancer Genet Cytogenet. 2006, 166: 166-172. 10.1016/j.cancergencyto.2005.11.005CrossRefPubMed
27.
Chêne L, Giroud C, Desgrandchamps F, Boccon-Gibod L, Cussenot O, Berthon P, Latil A: Extensive analysis of the 7q31 region in human prostate tumors supports TES as the best candidate tumor suppressor gene. Int J Cancer. 2004, 111: 798-804. 10.1002/ijc.20337CrossRefPubMed
28.
Zenklusen JC, Weintraub LA, Green ED: Construction of a high-resolution physical map of the approximate 1-Mb region of human chromosome 7q31.1-q31.2 harboring a putative tumor suppressor gene. Neoplasia. 1999, 1: 16-22. 10.1038/sj.neo.7900011PubMedCentralCrossRefPubMed
29.
Thomas NA, Choong DY, Jokubaitis VJ, Neville PJ, Campbell IG: Mutation of the ST7 tumor suppressor gene on 7q31.1 is rare in breast, ovarian and colorectal cancers. Nat Genet. 2001, 29: 379-380. 10.1038/ng784CrossRefPubMed
30.
Hughes KA, Hurlstone AF, Tobias ES, McFarlane R, Black DM: Absence of ST7 mutations in tumor-derived cell lines and tumors. Nat Genet. 2001, 29: 380-381. 10.1038/ng783CrossRefPubMed
31.
Drusco A, Zanesi N, Roldo C, Trapasso F, Farber JL, Fong LY, Croce CM: Knockout mice reveal a tumor suppressor function for Testin. Proc Natl Acad Sci USA. 2005, 102: 10947-10951. 10.1073/pnas.0504934102PubMedCentralCrossRefPubMed
32.
Sarti M, Sevignani C, Calin GA, Aqeilan R, Shimizu M, Pentimalli F, Picchio MC, Godwin A, Rosenberg A, Drusco A, Negrini M, Croce CM: Adenoviral transduction of TESTIN gene into breast and uterine cancer cell lines promotes apoptosis and tumor reduction in vivo. Clin Cancer Res. 2005, 11: 806-813.PubMed
33.
Garvalov BK, Higgins TE, Sutherland JD, Zettl M, Scaplehorn N, Köcher T, Piddini E, Griffiths G, Way M: The conformational state of Tes regulates its zyxin-dependent recruitment to focal adhesions. The Journal of Cell Biology. 2003, 161: 33-39. 10.1083/jcb.200211015PubMedCentralCrossRefPubMed
34.
Dallas PB, Gottardo NG, Firth MJ, Beesley AH, Hoffman , Terry PA, Freitas JR, Boag JM, Cummings AJ, Kees UR: Gene expression levels assessed by oligonucleotide microarray analysis and quantitative real-time RT-PCR -- how well do they correlate?. BMC Genomics. 2005, 6: 59-68. 10.1186/1471-2164-6-59PubMedCentralCrossRefPubMed
35.
Andersson A, Olofsson T, Lindgren D, Nilsson B, Ritz C, Edén P, Lassen C, Råde J, Fontes M, Mörse H, Heldrup J, Behrendtz M, Mitelman F, Höglund M, Johansson B, Fioretos T: Molecular signatures in childhood acute leukemia and their correlations to expression patterns in normal hematopoietic subpopulations. Proc Natl Acad Sci USA. 2005, 102: 19069-19074. 10.1073/pnas.0506637102PubMedCentralCrossRefPubMed
36.
Ross ME, Zhou X, Song G, Shurtleff SA, Girtman K, Williams WK, Liu HC, Mahfouz R, Raimondi SC, Lenny N, Patel A, Downing JR: Classification of pediatric acute lymphoblastic leukemia by gene expression profiling. Blood. 2003, 102: 2951-2959. 10.1182/blood-2003-01-0338CrossRefPubMed
37.
Coutts A, MacKenzie E, Griffith E, Black D: TES is a novel focal adhesion protein with a role in cell spreading. Journal of Cell Science. 2003, 116: 897-906. 10.1242/jcs.00278CrossRefPubMed
38.
Boëda B, Briggs DC, Higgins T, Garvalov BK, Fadden AJ, McDonald NQ, Way M: Tes, a specific Mena interacting partner, breaks the rules for EVH1 binding. Mol Cell. 2007, 28: 1071-1082. 10.1016/j.molcel.2007.10.033CrossRefPubMed
39.
Griffith E, Coutts AS, Black DM: Characterisation of chicken TES and its role in cell spreading and motility. Cell Motil Cytoskeleton. 2004, 57: 133-142. 10.1002/cm.10162CrossRefPubMed
40.
Griffith E, Coutts A, Black D: RNAi knockdown of the focal adhesion protein TES reveals its role in actin stress fibre organisation. Cell Motil Cytoskeleton. 2005, 60: 140-152. 10.1002/cm.20052CrossRefPubMed
41.
Nakata Y, Tomkowicz B, Gewirtz AM, Ptasznik A: Integrin inhibition through Lyn-dependent cross talk from CXCR4 chemokine receptors in normal human CD34+ marrow cells. Blood. 2006, 107: 4234-4239. 10.1182/blood-2005-08-3343PubMedCentralCrossRefPubMed
42.
Zhong Y, Zhu J, Wang Y, Zhou J, Ren K, Ding X, Zhang J: LIM domain protein TES changes its conformational states in different cellular compartments. Mol Cell Biochem. 2009, 320: 85-92. 10.1007/s11010-008-9901-7CrossRefPubMed
43.
Rabbitts TH: LMO T-cell translocation oncogenes typify genes activated by chromosomal translocations that alter transcription and developmental processes. Genes Dev. 1998, 12: 2651-2657. 10.1101/gad.12.17.2651CrossRefPubMed
44.
Howe SJ, Mansour MR, Schwarzwaelder K, Bartholomae C, Hubank M, Kempski H, Brugman MH, Pike-Overzet K, Chatters SJ, de Ridder D, Gilmour KC, Adams S, Thornhill SI, Parsley KL, Staal FJT, Gale RE, Linch DC, Bayford J, Brown L, Quaye M, Kinnon C, Ancliff P, Webb DK, Schmidt M, von Kalle C, Gaspar HR, Thrasher AJ: Insertional mutagenesis combined with acquired somatic mutations causes leukemogenesis following gene therapy of SCID-X1 patients. J Clin Invest. 2008, 118: 3143-3150. 10.1172/JCI35798PubMedCentralCrossRefPubMed
45.
Wadman IA, Osada H, Grütz GG, Agulnick AD, Westphal H, Forster A, Rabbitts TH: The LIM-only protein Lmo2 is a bridging molecule assembling an erythroid, DNA-binding complex which includes the TAL1, E47, GATA-1 and Ldb1/NLI proteins. EMBO J. 1997, 16: 3145-3157. 10.1093/emboj/16.11.3145PubMedCentralCrossRefPubMed
46.
Weeks R, Morison I: Detailed methylation analysis of CpG islands on human chromosome region 9p21. Genes Chromosomes Cancer. 2006, 45: 357-364. 10.1002/gcc.20297CrossRefPubMed
47.
Li LC, Dahiya R: MethPrimer: designing primers for methylation PCRs. Bioinformatics. 2002, 18: 1427-1431. 10.1093/bioinformatics/18.11.1427CrossRefPubMed
Metadata
Title
Silencing of TESTIN by dense biallelic promoter methylation is the most common molecular event in childhood acute lymphoblastic leukaemia
Authors
Robert J. Weeks
Ursula R. Kees
Sarah Song
Ian M. Morison
Publication date
01-12-2010
Publisher
BioMed Central
Published in
Molecular Cancer / Issue 1/2010
Electronic ISSN: 1476-4598
DOI
https://doi.org/10.1186/1476-4598-9-163

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