TY - JOUR SN - 1553-7390 UR - http://dx.doi.org/10.1371/journal.pgen.1000745 PB - PUBLIC LIBRARY SCIENCE ID - discovery358409 N2 - Wilms' tumour (WT) is a pediatric tumor of the kidney that arises via failure of the fetal developmental program. The absence of identifiable mutations in the majority of WTs suggests the frequent involvement of epigenetic aberrations in WT. We therefore conducted a genome-wide analysis of promoter hypermethylation in WTs and identified hypermethylation at chromosome 5q31 spanning 800 kilobases (kb) and more than 50 genes. The methylated genes all belong to alpha-, beta-, and gamma-protocadherin (PCDH) gene clusters (Human Genome Organization nomenclature PCDHA@, PCDHB@, and PCDHG@, respectively). This demonstrates that long-range epigenetic silencing (LRES) occurs in developmental tumors as well as in adult tumors. Bisulfite polymerase chain reaction analysis showed that PCDH hypermethylation is a frequent event found in all Wilms' tumor subtypes. Hypermethylation is concordant with reduced PCDH expression in tumors. WT precursor lesions showed no PCDH hypermethylation, suggesting that de novo PCDH hypermethylation occurs during malignant progression. Discrete boundaries of the PCDH domain are delimited by abrupt changes in histone modifications; unmethylated genes flanking the LRES are associated with permissive marks which are absent from methylated genes within the domain. Silenced genes are marked with non-permissive histone 3 lysine 9 dimethylation. Expression analysis of embryonic murine kidney and differentiating rat metanephric mesenchymal cells demonstrates that Pcdh expression is developmentally regulated and that Pcdhg@ genes are expressed in blastemal cells. Importantly, we show that PCDHs negatively regulate canonical Wnt signalling, as short-interfering RNA-induced reduction of PCDHG@ encoded proteins leads to elevated beta-catenin protein, increased beta-catenin/T-cell factor (TCF) reporter activity, and induction of Wnt target genes. Conversely, over-expression of PCDHs suppresses beta-catenin/TCF-reporter activity and also inhibits colony formation and growth of cancer cells in soft agar. Thus PCDHs are candidate tumor suppressors that modulate regulatory pathways critical in development and disease, such as canonical Wnt signaling. KW - GAMMA-PROTOCADHERINS KW - BREAST-CANCER KW - COLORECTAL-CANCER KW - SUPPRESSOR GENE KW - KIDNEY DEVELOPMENT KW - NEPHROGENIC RESTS KW - DNA METHYLATION KW - COMMON EVENT KW - LUNG-CANCER KW - CELL-LINE A1 - Dallosso, AR A1 - Hancock, AL A1 - Szemes, M A1 - Moorwood, K A1 - Chilukamarri, L A1 - Tsai, HH A1 - Sarkar, A A1 - Barasch, J A1 - Vuononvirta, R A1 - Jones, C A1 - Pritchard-Jones, K A1 - Royer-Pokora, B A1 - Lee, SB A1 - Owen, C A1 - Malik, S A1 - Feng, Y A1 - Frank, M A1 - Ward, A A1 - Brown, KW A1 - Malik, K JF - PLOS GENET Y1 - 2009/11/26/ AV - public VL - 5 TI - Frequent Long-Range Epigenetic Silencing of Protocadherin Gene Clusters on Chromosome 5q31 in Wilms' Tumor N1 - This is an open-access article distributed under the terms of the Creative Commons Public Domain declaration which stipulates that, once placed in the public domain, this work may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose. Funding: This work was supported by Cancer and Leukaemia in Childhood - Sargent, the Children's Leukaemia Trust, and Kidney Research UK. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. IS - 11 ER -