Methylation-sensitive high-resolution melting technology is a simple and sensitive method to detect germline epimutation of the MLH1 gene promoter
Abstract Background Lynch syndrome is an autosomal dominant cancer predisposition syndrome classically caused by germline mutations of the mismatch repair genes MLH1, MSH2, MSH6 and PMS2. Epimutation (also called germline hypermethylation) of the MLH1 gene promoter explains rare Lynch syndrome cases...
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BMC
2025-07-01
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| Series: | Clinical Epigenetics |
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| Online Access: | https://doi.org/10.1186/s13148-025-01904-1 |
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| author | Hélène Delhomelle Olfa Trabelsi-Grati Marie-Charlotte Villy Sabrina Ibadioune Frederic Maraone Mathieu Séné Bruno Buecher Emmanuelle Mouret-Fourme Marion Gauthier-Villars Faustine Johannes Lisa Golmard Anne Vincent-Salomon Eric Pasmant Julie Leclerc Michel Bahuau Ivan Bièche Chrystelle Colas |
| author_facet | Hélène Delhomelle Olfa Trabelsi-Grati Marie-Charlotte Villy Sabrina Ibadioune Frederic Maraone Mathieu Séné Bruno Buecher Emmanuelle Mouret-Fourme Marion Gauthier-Villars Faustine Johannes Lisa Golmard Anne Vincent-Salomon Eric Pasmant Julie Leclerc Michel Bahuau Ivan Bièche Chrystelle Colas |
| author_sort | Hélène Delhomelle |
| collection | DOAJ |
| description | Abstract Background Lynch syndrome is an autosomal dominant cancer predisposition syndrome classically caused by germline mutations of the mismatch repair genes MLH1, MSH2, MSH6 and PMS2. Epimutation (also called germline hypermethylation) of the MLH1 gene promoter explains rare Lynch syndrome cases. To date, there is no recommendation regarding the techniques to be used to detect this epimutation, nor any clinical criteria for when it should be investigated. We present a retrospective study involving 73 patients whose Lynch syndrome-related tumours exhibited MLH1 hypermethylation, and compare the methylation-sensitive high-resolution melting (MS-HRM) technology to the gold standard method of pyrosequencing, in view of performing routine MLH1 epimutation testing. We further ascertained epimutation of MLH1 by MS-HRM from a control group of 25 patients with known MMR germline pathogenic variant. Results MLH1 epimutation was detected in 6.8% of the comprehensive cohort (5/73). MS-HRM detected one positive case previously identified by pyrosequencing, along with two missed cases at very low allelic frequencies and allowed the identification of two additional positive patients, subsequently confirmed by pyrosequencing. No patients in the control group were found to have MLH1 epimutation. Conclusion MS-HRM is a simple and sensitive real-time PCR method amenable to routinely detect MLH1 epimutation, with high sensitivity. Identifying patients with MLH1 epimutation is of paramount importance for patient care and genetic counselling. We further recommend performing this analysis systematically in discrete clinical settings herewith specified. |
| format | Article |
| id | doaj-art-aeec04fcb26d4ea8a347cd3c76150b0e |
| institution | DOAJ |
| issn | 1868-7083 |
| language | English |
| publishDate | 2025-07-01 |
| publisher | BMC |
| record_format | Article |
| series | Clinical Epigenetics |
| spelling | doaj-art-aeec04fcb26d4ea8a347cd3c76150b0e2025-08-20T03:03:41ZengBMCClinical Epigenetics1868-70832025-07-0117111210.1186/s13148-025-01904-1Methylation-sensitive high-resolution melting technology is a simple and sensitive method to detect germline epimutation of the MLH1 gene promoterHélène Delhomelle0Olfa Trabelsi-Grati1Marie-Charlotte Villy2Sabrina Ibadioune3Frederic Maraone4Mathieu Séné5Bruno Buecher6Emmanuelle Mouret-Fourme7Marion Gauthier-Villars8Faustine Johannes9Lisa Golmard10Anne Vincent-Salomon11Eric Pasmant12Julie Leclerc13Michel Bahuau14Ivan Bièche15Chrystelle Colas16Department of Genetics, Institut CurieDepartment of Genetics, Institut CurieDepartment of Genetics, Institut CurieDepartment of Genetics, Institut CurieDepartment of Genetics, Institut CurieDepartment of Genetics, Institut CurieDepartment of Genetics, Institut CurieDepartment of Genetics, Institut CurieDepartment of Genetics, Institut CurieService de Génétique Clinique, Centre Hospitalier IntercommunalDepartment of Genetics, Institut CurieDepartment of Pathology-Genetics and Immunology, Institut CurieDepartment of Genetics, Institut CurieMolecular Oncogenetics, Department of Biochemistry and Molecular Biology, CHU LilleService de Génétique Clinique, Centre Hospitalier IntercommunalDepartment of Genetics, Institut CurieDepartment of Genetics, Institut CurieAbstract Background Lynch syndrome is an autosomal dominant cancer predisposition syndrome classically caused by germline mutations of the mismatch repair genes MLH1, MSH2, MSH6 and PMS2. Epimutation (also called germline hypermethylation) of the MLH1 gene promoter explains rare Lynch syndrome cases. To date, there is no recommendation regarding the techniques to be used to detect this epimutation, nor any clinical criteria for when it should be investigated. We present a retrospective study involving 73 patients whose Lynch syndrome-related tumours exhibited MLH1 hypermethylation, and compare the methylation-sensitive high-resolution melting (MS-HRM) technology to the gold standard method of pyrosequencing, in view of performing routine MLH1 epimutation testing. We further ascertained epimutation of MLH1 by MS-HRM from a control group of 25 patients with known MMR germline pathogenic variant. Results MLH1 epimutation was detected in 6.8% of the comprehensive cohort (5/73). MS-HRM detected one positive case previously identified by pyrosequencing, along with two missed cases at very low allelic frequencies and allowed the identification of two additional positive patients, subsequently confirmed by pyrosequencing. No patients in the control group were found to have MLH1 epimutation. Conclusion MS-HRM is a simple and sensitive real-time PCR method amenable to routinely detect MLH1 epimutation, with high sensitivity. Identifying patients with MLH1 epimutation is of paramount importance for patient care and genetic counselling. We further recommend performing this analysis systematically in discrete clinical settings herewith specified.https://doi.org/10.1186/s13148-025-01904-1Lynch SyndromeMS-HRMPyrosequencingMLH1 gene promoterHypermethylationEpimutation |
| spellingShingle | Hélène Delhomelle Olfa Trabelsi-Grati Marie-Charlotte Villy Sabrina Ibadioune Frederic Maraone Mathieu Séné Bruno Buecher Emmanuelle Mouret-Fourme Marion Gauthier-Villars Faustine Johannes Lisa Golmard Anne Vincent-Salomon Eric Pasmant Julie Leclerc Michel Bahuau Ivan Bièche Chrystelle Colas Methylation-sensitive high-resolution melting technology is a simple and sensitive method to detect germline epimutation of the MLH1 gene promoter Clinical Epigenetics Lynch Syndrome MS-HRM Pyrosequencing MLH1 gene promoter Hypermethylation Epimutation |
| title | Methylation-sensitive high-resolution melting technology is a simple and sensitive method to detect germline epimutation of the MLH1 gene promoter |
| title_full | Methylation-sensitive high-resolution melting technology is a simple and sensitive method to detect germline epimutation of the MLH1 gene promoter |
| title_fullStr | Methylation-sensitive high-resolution melting technology is a simple and sensitive method to detect germline epimutation of the MLH1 gene promoter |
| title_full_unstemmed | Methylation-sensitive high-resolution melting technology is a simple and sensitive method to detect germline epimutation of the MLH1 gene promoter |
| title_short | Methylation-sensitive high-resolution melting technology is a simple and sensitive method to detect germline epimutation of the MLH1 gene promoter |
| title_sort | methylation sensitive high resolution melting technology is a simple and sensitive method to detect germline epimutation of the mlh1 gene promoter |
| topic | Lynch Syndrome MS-HRM Pyrosequencing MLH1 gene promoter Hypermethylation Epimutation |
| url | https://doi.org/10.1186/s13148-025-01904-1 |
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