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Germline Mutations of the hMLH1 and hMSH2 Mismatch Repair Genes in Belgian Hereditary Nonpolyposis Colon Cancer (HNPCC) Patients

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Abstract

Background: Hereditary nonpolyposis colon cancer (HNPCC-Lynch syndrome) is caused by mutations in genes involved in DNA mismatch repair (MMR), mostly in the hMLH1 and hMSH2 genes. The mutation spectrum in the Belgian population is still poorly documented. Aim: To report our experience on the mutation screening in Belgian familial colorectal cancer (CRC) patients, including the investigation of the pathogenicity of the missense and splice mutations. To increase the mutation detection rate by selecting the target population. Methods: Two hundred and twenty five Belgian patients with familial clustering of CRC were genetically tested. Point mutations in the hMLH1 and hMSH2 genes were screened by denaturing gradient gel electrophoresis (DGGE) followed by direct sequencing. Genomic deletions and duplications were assessed by multiplex ligase dependent probe amplification (MLPA) and multiplex PCR. Missense mutations were examined for pathogenicity by means of cosegregation of the mutation with the disease, microsatellite instability (MSI) in tumors, immunohistochemical staining of tumors and determination of the population frequency of the particular mutation. Results: Twenty five pathogenic mutations were identified from which 16 were novel: 7 frameshifts, one in frame deletion, 5 genomic deletions, 5 splice defects, 4 nonsense (stop) mutations and 3 missense mutations which were classified as pathogenic (out of 10 missense mutations). In retrospect, a mutation detection rate of 71% was obtained if MSI was used as a supplementary selection criterion in addition to familial clustering. Conclusion: Different types of pathogenic mutations in the hMLH1 and hMSH2 genes were identified in a Belgian CRC group with familial clustering. The mutation detection yield drastically increased by preliminar selection of those familial CRC patients with a microsatellite instable tumor. Considerable attention went to the assessment of the pathogenicity of the missense mutations. In practice, the cosegregation with the disease was the most relevant criterion.

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References

  1. CR Boland (2005) ArticleTitleEvolution of the nomenclature for the hereditary colorectal cancer syndromes Fam Cancer 4 211–8 Occurrence Handle16136380 Occurrence Handle10.1007/s10689-004-4489-x

    Article  PubMed  Google Scholar 

  2. P Peltomäki (2005) ArticleTitleLynch syndrome genes Fam Cancer 4 227–32 Occurrence Handle16136382 Occurrence Handle10.1007/s10689-004-7993-0 Occurrence Handle1:CAS:528:DC%2BD2MXps1alsbs%3D

    Article  PubMed  CAS  Google Scholar 

  3. HFA Vasen J-P Mecklin PM Khan HT Lynch (1991) ArticleTitleThe International Collaborative Group on Hereditary Non-Polyposis Colorectal Cancer (ICG-HNPCC) Dis Colon Rectum 34 424–5 Occurrence Handle2022152 Occurrence Handle1:STN:280:DyaK3M3hsF2ksw%3D%3D Occurrence Handle10.1007/BF02053699

    Article  PubMed  CAS  Google Scholar 

  4. JT Wijnen HFA Vasen PM Khan et al. (1998) ArticleTitleClinical findings with implications for genetic testing in families with clustering of colorectal cancer N Engl J Med 339 511–8 Occurrence Handle9709044 Occurrence Handle1:STN:280:DyaK1czmtlKlsQ%3D%3D Occurrence Handle10.1056/NEJM199808203390804

    Article  PubMed  CAS  Google Scholar 

  5. HFA Vasen (2005) ArticleTitleClinical description of the Lynch syndrome [hereditary nonpolyposis colorectal cancer (HNPCC)] Familial Cancer 4 219–25 Occurrence Handle16136381 Occurrence Handle1:STN:280:DC%2BD2MvmslKrsA%3D%3D Occurrence Handle10.1007/s10689-004-3906-5

    Article  PubMed  CAS  Google Scholar 

  6. S Tejpar E Cutsem ParticleVan (1998) ArticleTitleHereditary non-polyposis colorectal cancer Progress in colorectal cancer 2 1–2

    Google Scholar 

  7. J Wijnen H Vasen PM Khan et al. (1995) ArticleTitleSeven new mutations in hMSH2, an HNPCC gene, identified by denaturing gradient-gel electrophoresis Am J Hum Genet 56 1060–6 Occurrence Handle7726159 Occurrence Handle1:STN:280:DyaK2M3ktlahsg%3D%3D

    PubMed  CAS  Google Scholar 

  8. J Wijnen PM Khan H Vasen et al. (1996) ArticleTitleMajority of hMLH1 mutations responsible for hereditary nonpolyposis colorectal cancer cluster at the exonic region 15–16 Am J Hum Genet 58 300–7 Occurrence Handle8571956 Occurrence Handle1:CAS:528:DyaK28XhsFWitbo%3D

    PubMed  CAS  Google Scholar 

  9. W Dietmaier S Wallinger T Bocker et al. (1997) ArticleTitleDiagnostic microsatellite instability: definition and correlation with mismatch repair protein expression Cancer Res 57 4749–56 Occurrence Handle9354436 Occurrence Handle1:CAS:528:DyaK2sXnt12gurc%3D

    PubMed  CAS  Google Scholar 

  10. CR Boland SN Thibodeau SR Hamilton et al. (1998) ArticleTitleA National Cancer Institute Workshop on microsatellite instability for cancer detection and familial predisposition: development of international criteria for the determination of microsatellite instability in colorectal cancer Cancer Res 58 5248–57 Occurrence Handle9823339 Occurrence Handle1:CAS:528:DyaK1cXnsFSmtbk%3D

    PubMed  CAS  Google Scholar 

  11. JM Cunningham ER Christensen DJ Tester et al. (1998) ArticleTitleHypermethylation of the hMLH1 promoter in colon cancer with microsatellite instability Cancer Res 58 3455–60 Occurrence Handle9699680 Occurrence Handle1:CAS:528:DyaK1cXltFyis7Y%3D

    PubMed  CAS  Google Scholar 

  12. NJ Frogatt JA Joyce R Davies et al. (1995) ArticleTitleA Frequent hMSH2 mutation in hereditary non-polyposis cancer syndrome The Lancet 345 727 Occurrence Handle10.1016/S0140-6736(95)90900-1

    Article  Google Scholar 

  13. JJP Gille FBL Hogervorst G Pals et al. (2002) ArticleTitleGenomic deletions of MSH2 and MLH1 in colorectal cancer families detected by a novel mutation detection approach Br J Cancer 87 892–7 Occurrence Handle12373605 Occurrence Handle1:CAS:528:DC%2BD38Xns1GntL0%3D Occurrence Handle10.1038/sj.bjc.6600565

    Article  PubMed  CAS  Google Scholar 

  14. JP Schouten CJ Mc Elgunn R Waaijer (2002) ArticleTitleRelative quantification of 40 nucleic acid sequences by multiplex ligation-dependent probe amplification Nucleic Acids Res 30 e57 Occurrence Handle12060695 Occurrence Handle10.1093/nar/gnf056

    Article  PubMed  Google Scholar 

  15. F Charbonnier G Raux Q Wang et al. (2000) ArticleTitleDetection of exon deletions and duplications of the mismatch repair genes in hereditary nonpolyposis colorectal cancer families using multiplex polymerase chain reaction of short fluorescent fragments Cancer Res 60 2760–63 Occurrence Handle10850409 Occurrence Handle1:CAS:528:DC%2BD3cXjvFOls7s%3D

    PubMed  CAS  Google Scholar 

  16. F Charbonnier S Olschwang Q Wang et al. (2002) ArticleTitleMSH2 in contrast to MLH1 and MSH6 is frequently inactivated by exonic and promoter rearrangements in hereditary nonpolyposis colorectal cancer Cancer Res 62 848–53 Occurrence Handle11830542 Occurrence Handle1:CAS:528:DC%2BD38Xht1Kru7Y%3D

    PubMed  CAS  Google Scholar 

  17. Raedle J, Brieger A, Trojan J, et al. Detection of hMLH1 and hMSH2 germline mutations in hereditary nonpolyposis colorectal cancer (HNPCC) using base excision sequence scanning anlysis. Gastrointest Oncol 1999; 116: A489

  18. B Liu RE Parsons SR Hamilton et al. (1994) ArticleTitlehMSH2 mutations in hereditary nonpolyposis colorectal cancer kindreds Cancer Res 54 4590–4 Occurrence Handle8062247 Occurrence Handle1:CAS:528:DyaK2cXlvFOgu7Y%3D

    PubMed  CAS  Google Scholar 

  19. JM Buerstedde P Alday J Torhorst et al. (1995) ArticleTitleDetection of new mutations in six out of 10 Swiss HNPCC families by genomic sequencing of the hMSH2 and hMLH1 genes J Med Genet 32 909–1012 Occurrence Handle8592341 Occurrence Handle1:CAS:528:DyaK28Xis1GhsA%3D%3D Occurrence Handle10.1136/jmg.32.11.909

    Article  PubMed  CAS  Google Scholar 

  20. E Kondo H Suzuki A Horii et al. (2003) ArticleTitleA yeast two-hybrid assay provides a simple way to evaluate the vast majority of hMLH1 germ-line mutations Cancer Res 63 3302–8 Occurrence Handle12810663 Occurrence Handle1:CAS:528:DC%2BD3sXks1ehsr4%3D

    PubMed  CAS  Google Scholar 

  21. MF Kane M Loda GM Gaida et al. (1997) ArticleTitleMethylation of the hMLH1 promoter correlates with lack of expression of hMLH1 in sporadic colon tumors and mismatch repair-defective human tumor cell lines Cancer Res 57 808–11 Occurrence Handle9041175 Occurrence Handle1:CAS:528:DyaK2sXhs1Kjtbs%3D

    PubMed  CAS  Google Scholar 

  22. M Genuardi S Carrara M Anti et al. (1999) ArticleTitleAssessment of pathogenicity criteria for constitutional missense mutations of the hereditary nonpolyposis colorectal cancer genes MLH1 and MSH2 Eur J Hum Genet 7 778–82 Occurrence Handle10573010 Occurrence Handle1:CAS:528:DyaK1MXnvFSju70%3D Occurrence Handle10.1038/sj.ejhg.5200363

    Article  PubMed  CAS  Google Scholar 

  23. S Syngal EA Fox C Li et al. (1999) ArticleTitleInterpretation of genetic test results for hereditay nonpolyposis colorectal cancer JAMA 282 247–53 Occurrence Handle10422993 Occurrence Handle1:STN:280:DyaK1MzkvFymtA%3D%3D Occurrence Handle10.1001/jama.282.3.247

    Article  PubMed  CAS  Google Scholar 

  24. M Cravo AJ Afonso P Lage et al. (2002) ArticleTitlePathogenicity of missense and splice site mutations in hMSH2 and hMLH1 mismatch repair genes: implications for genetic testing Gut 50 405–12 Occurrence Handle11839723 Occurrence Handle1:CAS:528:DC%2BD38XitFegsLg%3D Occurrence Handle10.1136/gut.50.3.405

    Article  PubMed  CAS  Google Scholar 

  25. H Shimodaira N Filosi H Shibata et al. (1998) ArticleTitleFunctional analysis of human MLH1 mutations in Saccharomyces cerevisiae Nat Genet 19 384–9 Occurrence Handle9697702 Occurrence Handle1:CAS:528:DyaK1cXlt1Cisrk%3D Occurrence Handle10.1038/1277

    Article  PubMed  CAS  Google Scholar 

  26. S Guerette S Acharya R Fishel (1999) ArticleTitleThe interaction of the human Mut L homologues in hereditary nonpolyposis colon cancer J Biol Chem 274 6336–41 Occurrence Handle10.1074/jbc.274.10.6336

    Article  Google Scholar 

  27. AR Ellison J Lofing GA Bitter et al. (2001) ArticleTitleFunctional analysis of human MLH1 and MSH2 missense variants and hybrid human–yeast MLH1 proteins in Saccharomyces cerevisiae Hum Mol Genet 10 1889–900 Occurrence Handle11555625 Occurrence Handle1:CAS:528:DC%2BD3MXntlSgsb4%3D Occurrence Handle10.1093/hmg/10.18.1889

    Article  PubMed  CAS  Google Scholar 

  28. M Nyström-Lahti C Perrera M Räschle et al. (2002) ArticleTitleFunctional analysis of MLH1 mutations linked to hereditary nonpolyposis colon cancer Genes Chromosomes Cancer 33 160–7 Occurrence Handle11793442 Occurrence Handle10.1002/gcc.1225 Occurrence Handle1:CAS:528:DC%2BD38XntFWmtg%3D%3D

    Article  PubMed  CAS  Google Scholar 

  29. J Trojan S Zeuzem A Randolph et al. (2002) ArticleTitleFunctional analysis of hMLH1 variants and HNPCC-related mutations using a human expression system Gastroenterology 122 211–9 Occurrence Handle11781295 Occurrence Handle1:CAS:528:DC%2BD38Xmt1Crsg%3D%3D Occurrence Handle10.1053/gast.2002.30296

    Article  PubMed  CAS  Google Scholar 

  30. A Wagner A Barrows JT Wijnen et al. (2003) ArticleTitleMolecular analysis of hereditary colorectal cancer in the United States: High mutation detection rate among clinically selected families and characterization of an American founder genomic deletion of the MSH2 gene Am J Hum Genet 72 1088–100 Occurrence Handle12658575 Occurrence Handle1:CAS:528:DC%2BD3sXjslagtbo%3D Occurrence Handle10.1086/373963

    Article  PubMed  CAS  Google Scholar 

  31. Nakagawa H, Hampel H, de la Chapelle A. Identification an characterization of genomic rearrangements of MSH2 and MLH1 in Lynch syndrome (HNPCC) by novel techniques. Hum Mutat 2003; Mutation in Brief #642 online

  32. CF Taylor RS Charlton J Burn et al. (2003) ArticleTitleGenomic deletions in MSH2 or MLH1 are a frequent cause of hereditary non-polyposis colorectal cancer: Identification of novel and recurrent deletions by MLPA Hum Mut 22 428–33 Occurrence Handle14635101 Occurrence Handle1:CAS:528:DC%2BD2cXps1Kl Occurrence Handle10.1002/humu.10291

    Article  PubMed  CAS  Google Scholar 

  33. C Lamberti R Kruse C Ruelfs et al. (1999) ArticleTitleMicrosatellite instability – a useful diagnostic tool to select patients at high risk for hereditary non-polyposis colorectal cancer: a study in different groups of patients with colorectal cancer Gut 44 839–43 Occurrence Handle10323887 Occurrence Handle1:STN:280:DyaK1M3lslaqtw%3D%3D Occurrence Handle10.1136/gut.44.6.839

    Article  PubMed  CAS  Google Scholar 

  34. T Liu S Wahlberg E Burek et al. (2000) ArticleTitleMicrosatellite instability as a predictor of a mutation in a DNA mismatch repair gene in familial colorectal cancer Genes Chromosomes Cancer 27 17–25 Occurrence Handle10564582 Occurrence Handle1:CAS:528:DC%2BD3cXksFym Occurrence Handle10.1002/(SICI)1098-2264(200001)27:1<17::AID-GCC3>3.0.CO;2-Y

    Article  PubMed  CAS  Google Scholar 

  35. JM Wheeler A Loukola LA Aaltonen et al. (2000) ArticleTitleThe role of hypermethylation of the hMLH1 promoter region in HNPCC versus MSI+sporadic colorectal cancers J Med Genet 37 588–92 Occurrence Handle10922385 Occurrence Handle1:CAS:528:DC%2BD3cXmtFOhtb0%3D Occurrence Handle10.1136/jmg.37.8.588

    Article  PubMed  CAS  Google Scholar 

  36. A Umar R Boland JP Terdiman et al. (2004) ArticleTitleRevised Bethesda guidelines for hereditary nonpolyposis colorectal cancer (lynch syndrome) and microsatellite instability J Natl Cancer Inst 96 261–8 Occurrence Handle14970275 Occurrence Handle1:CAS:528:DC%2BD2cXhsV2qtLk%3D Occurrence Handle10.1093/jnci/djh034

    Article  PubMed  CAS  Google Scholar 

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Authors and Affiliations

  1. Center for Human Genetics, University Hospital of Leuven, Leuven, Belgium

    M. Spaepen, B. Vankeirsbilck, S. Van Opstal, S. Tejpar, E. Legius & G. Matthijs

  2. Department of Gastroenterology, University Hospital of Leuven, Leuven, Belgium

    S. Tejpar & E. Van Cutsem

  3. Department of Pathological Anatomy of Leuven, University Hospital of Leuven, Leuven, Belgium

    K. Geboes

  4. Center for Human Genetics, University of Leuven, Herestraat, 49, 3000, Leuven, Belgium

    M. Spaepen

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  1. M. Spaepen
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Correspondence to M. Spaepen.

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Spaepen, M., Vankeirsbilck, B., Van Opstal, S. et al. Germline Mutations of the hMLH1 and hMSH2 Mismatch Repair Genes in Belgian Hereditary Nonpolyposis Colon Cancer (HNPCC) Patients. Familial Cancer 5, 179–189 (2006). https://doi.org/10.1007/s10689-005-5958-6

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