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Heritable germline epimutation of MSH2 in a family with hereditary nonpolyposis colorectal cancer

Abstract

Epimutations in the germline, such as methylation of the MLH1 gene, may contribute to hereditary cancer syndrome in human, but their transmission to offspring has never been documented. Here we report a family with inheritance, in three successive generations, of germline allele-specific and mosaic hypermethylation of the MSH2 gene, without evidence of DNA mismatch repair gene mutation. Three siblings carrying the germline methylation developed early-onset colorectal or endometrial cancers, all with microsatellite instability and MSH2 protein loss. Clonal bisulfite sequencing and pyrosequencing showed different methylation levels in different somatic tissues, with the highest level recorded in rectal mucosa and colon cancer tissue, and the lowest in blood leukocytes. This mosaic state of germline methylation with different tissue distribution could act as the first hit and provide a mechanism for genetic disease inheritance that may deviate from the mendelian pattern and be overlooked in conventional leukocyte-based genetic diagnosis strategy.

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Figure 1: Family pedigree and immunohistochemical staining for DNA mismatch repair proteins in the tumors.
Figure 2: Methylation analysis of the MSH2 promoter and association with a specific allele of chromosome 2.
Figure 3: Quantitative estimates of methylation percentage in different somatic tissues from the germline epimutation carriers and control population by bisulfite conversion of DNA and pyrosequencing.

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Acknowledgements

We thank V. Yu, K.C. Lee, J. Ho, B. Leung and E. Chan for sample coordination and clinical care; W.M. Ho and C.W. Wong for technical assistance; J. Sham and The Genome Research Centre of The University of Hong Kong for support and P.C. Sham for help with linkage analysis. This work was supported by research grants from the Hong Kong Cancer Fund and the Kadoorie Charitable Foundation.

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Correspondence to Siu Tsan Yuen or Suet Yi Leung.

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The authors declare no competing financial interests.

Supplementary information

Supplementary Fig. 1

Immunohistochemical staining for MSH2. (PDF 77 kb)

Supplementary Fig. 2

Methylation-specific PCR screening for MSH2 promoter methylation in all family members. (PDF 369 kb)

Supplementary Fig. 3

Representative pyrosequencing chromatogram of the MSH2 promoter after bisulfite conversion and amplification by NP2, encompassing CpG sites 8-10. (PDF 96 kb)

Supplementary Table 1

Details of clinical information, MSI analysis, and MSH2 and MLH1 immunostaining results for the current family. (PDF 60 kb)

Supplementary Table 2

Clonal bisulfite allelic sequencing of blood leukocyte DNA from three healthy blood donors with a G/T polymorphism in SNP-433, using methylation-unbiased primers NP1 and NP2. (PDF 14 kb)

Supplementary Methods (PDF 27 kb)

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Chan, T., Yuen, S., Kong, C. et al. Heritable germline epimutation of MSH2 in a family with hereditary nonpolyposis colorectal cancer. Nat Genet 38, 1178–1183 (2006). https://doi.org/10.1038/ng1866

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