{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,24]],"date-time":"2026-06-24T02:39:43Z","timestamp":1782268783178,"version":"3.54.5"},"reference-count":31,"publisher":"Springer Science and Business Media LLC","issue":"3","license":[{"start":{"date-parts":[[2025,7,26]],"date-time":"2025-07-26T00:00:00Z","timestamp":1753488000000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/www.springernature.com\/gp\/researchers\/text-and-data-mining"},{"start":{"date-parts":[[2025,7,26]],"date-time":"2025-07-26T00:00:00Z","timestamp":1753488000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/www.springernature.com\/gp\/researchers\/text-and-data-mining"}],"funder":[{"name":"USA National Cancer Institute","award":["R01CA218342"],"award-info":[{"award-number":["R01CA218342"]}]},{"name":"Spanish Ministry of Science","award":["PID2019-111254RB-I00 and PID2023-151585OB-I00"],"award-info":[{"award-number":["PID2019-111254RB-I00 and PID2023-151585OB-I00"]}]},{"name":"Instituto de Salud Carlos III and European Regional Development FEDER","award":["PI19\/01336"],"award-info":[{"award-number":["PI19\/01336"]}]}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Familial Cancer"],"DOI":"10.1007\/s10689-025-00482-8","type":"journal-article","created":{"date-parts":[[2025,7,26]],"date-time":"2025-07-26T04:12:51Z","timestamp":1753503171000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["MLH1 c.27G>A (p.Arg9=) is a synonymous likely\/pathogenic variant underlying variably mosaic constitutional MLH1 methylation in Lynch syndrome"],"prefix":"10.1007","volume":"24","author":[{"given":"Rocio","family":"Alvarez","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Paula","family":"Climent-Cant\u00f3","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"GiWon","family":"Shin","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Francesca Paola","family":"Aguirre","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lisa","family":"Zhou","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dennis J.","family":"Hazelett","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Brent K.","family":"Larson","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Covadonga","family":"Vara","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gabriel","family":"Capell\u00e1","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"V\u00edctor Lorca","family":"Castellanos","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Pilar Garre","family":"Rubio","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fran\u00e7oise","family":"Desseigne","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hanlee","family":"Ji","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jackie","family":"Cook","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Miranda","family":"Durkie","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marta","family":"Pineda","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Julie","family":"Leclerc","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3603-2309","authenticated-orcid":false,"given":"Megan P.","family":"Hitchins","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2025,7,26]]},"reference":[{"issue":"4\u20135","key":"482_CR1","doi-asserted-by":"publisher","first-page":"181","DOI":"10.1155\/2004\/460240","volume":"20","author":"HT Lynch","year":"2004","unstructured":"Lynch HT, Lynch JF (2004) Lynch syndrome: history and current status. Dis Markers 20(4\u20135):181\u2013198","journal-title":"Dis Markers"},{"issue":"1 Pt 1","key":"482_CR2","doi-asserted-by":"publisher","first-page":"191","DOI":"10.1158\/1078-0432.CCR-1118-3","volume":"10","author":"G Deng","year":"2004","unstructured":"Deng G et al (2004) BRAF mutation is frequently present in sporadic colorectal cancer with methylated hMLH1, but not in hereditary nonpolyposis colorectal cancer. Clin Cancer Res 10(1 Pt 1):191\u2013195","journal-title":"Clin Cancer Res"},{"issue":"3","key":"482_CR3","doi-asserted-by":"publisher","first-page":"301","DOI":"10.1007\/s10689-007-9124-1","volume":"6","author":"MB Loughrey","year":"2007","unstructured":"Loughrey MB et al (2007) Incorporation of somatic BRAF mutation testing into an algorithm for the investigation of hereditary non-polyposis colorectal cancer. Fam Cancer 6(3):301\u2013310","journal-title":"Fam Cancer"},{"issue":"15","key":"482_CR4","first-page":"3455","volume":"58","author":"JM Cunningham","year":"1998","unstructured":"Cunningham JM et al (1998) Hypermethylation of the hMLH1 promoter in colon cancer with microsatellite instability. Cancer Res 58(15):3455\u20133460","journal-title":"Cancer Res"},{"key":"482_CR5","doi-asserted-by":"publisher","first-page":"129","DOI":"10.1016\/j.ygyno.2023.02.017","volume":"171","author":"MP Hitchins","year":"2023","unstructured":"Hitchins MP et al (2023) MLH1-methylated endometrial cancer under 60 years of age as the \u201csentinel\u201d cancer in female carriers of high-risk constitutional MLH1 epimutation. Gynecol Oncol 171:129\u2013140","journal-title":"Gynecol Oncol"},{"issue":"7","key":"482_CR6","doi-asserted-by":"publisher","first-page":"743","DOI":"10.6004\/jnccn.2023.7020","volume":"21","author":"MP Hitchins","year":"2023","unstructured":"Hitchins MP et al (2023) Constitutional MLH1 methylation is a major contributor to mismatch repair-deficient, MLH1-methylated colorectal cancer in patients aged 55 years and younger. J Natl Compr Canc Netw 21(7):743-752 e11","journal-title":"J Natl Compr Canc Netw"},{"issue":"5","key":"482_CR7","doi-asserted-by":"publisher","first-page":"1392","DOI":"10.1053\/j.gastro.2005.09.003","volume":"129","author":"M Hitchins","year":"2005","unstructured":"Hitchins M et al (2005) MLH1 germline epimutations as a factor in hereditary nonpolyposis colorectal cancer. Gastroenterology 129(5):1392\u20131399","journal-title":"Gastroenterology"},{"issue":"10","key":"482_CR8","doi-asserted-by":"publisher","first-page":"625","DOI":"10.1038\/nrc4001","volume":"15","author":"MP Hitchins","year":"2015","unstructured":"Hitchins MP (2015) Constitutional epimutation as a mechanism for cancer causality and heritability? Nat Rev Cancer 15(10):625\u2013634","journal-title":"Nat Rev Cancer"},{"issue":"7","key":"482_CR9","doi-asserted-by":"publisher","first-page":"697","DOI":"10.1056\/NEJMoa064522","volume":"356","author":"MP Hitchins","year":"2007","unstructured":"Hitchins MP et al (2007) Inheritance of a cancer-associated MLH1 germ-line epimutation. N Engl J Med 356(7):697\u2013705","journal-title":"N Engl J Med"},{"issue":"7","key":"482_CR10","doi-asserted-by":"publisher","first-page":"953","DOI":"10.1001\/jamaoncol.2015.1484","volume":"1","author":"MA Sloane","year":"2015","unstructured":"Sloane MA et al (2015) Mosaic epigenetic inheritance as a cause of early-onset colorectal cancer. JAMA Oncol 1(7):953\u2013957","journal-title":"JAMA Oncol"},{"issue":"2","key":"482_CR11","doi-asserted-by":"publisher","first-page":"200","DOI":"10.1016\/j.ccr.2011.07.003","volume":"20","author":"MP Hitchins","year":"2011","unstructured":"Hitchins MP et al (2011) Dominantly inherited constitutional epigenetic silencing of MLH1 in a cancer-affected family is linked to a single nucleotide variant within the 5\u2019UTR. Cancer Cell 20(2):200\u2013213","journal-title":"Cancer Cell"},{"issue":"5","key":"482_CR12","doi-asserted-by":"publisher","first-page":"617","DOI":"10.1038\/ejhg.2013.200","volume":"22","author":"CT Kwok","year":"2014","unstructured":"Kwok CT et al (2014) The MLH1 c.-27C>A and c.85G>T variants are linked to dominantly inherited MLH1 epimutation and are borne on a European ancestral haplotype. Eur J Hum Genet 22(5):617\u2013624","journal-title":"Eur J Hum Genet"},{"issue":"8","key":"482_CR13","doi-asserted-by":"publisher","first-page":"513","DOI":"10.1136\/jmedgenet-2011-100050","volume":"48","author":"M Morak","year":"2011","unstructured":"Morak M et al (2011) Biallelic MLH1 SNP cDNA expression or constitutional promoter methylation can hide genomic rearrangements causing Lynch syndrome. J Med Genet 48(8):513\u2013519","journal-title":"J Med Genet"},{"issue":"1","key":"482_CR14","doi-asserted-by":"publisher","first-page":"25","DOI":"10.1038\/gim.2012.91","volume":"15","author":"RL Ward","year":"2013","unstructured":"Ward RL et al (2013) Identification of constitutional MLH1 epimutations and promoter variants in colorectal cancer patients from the Colon Cancer Family Registry. Genet Med 15(1):25\u201335","journal-title":"Genet Med"},{"issue":"12","key":"482_CR15","doi-asserted-by":"publisher","first-page":"1589","DOI":"10.1038\/gim.2018.47","volume":"20","author":"J Leclerc","year":"2018","unstructured":"Leclerc J et al (2018) Diversity of genetic events associated with MLH1 promoter methylation in Lynch syndrome families with heritable constitutional epimutation. Genet Med 20(12):1589\u20131599","journal-title":"Genet Med"},{"issue":"4","key":"482_CR16","doi-asserted-by":"publisher","first-page":"240","DOI":"10.1136\/jmedgenet-2017-104744","volume":"55","author":"M Morak","year":"2018","unstructured":"Morak M et al (2018) Comprehensive analysis of the MLH1 promoter region in 480 patients with colorectal cancer and 1150 controls reveals new variants including one with a heritable constitutional MLH1 epimutation. J Med Genet 55(4):240\u2013248","journal-title":"J Med Genet"},{"issue":"4","key":"482_CR17","doi-asserted-by":"publisher","first-page":"452","DOI":"10.1093\/carcin\/bgv015","volume":"36","author":"G Cini","year":"2015","unstructured":"Cini G et al (2015) Concomitant mutation and epimutation of the MLH1 gene in a Lynch syndrome family. Carcinogenesis 36(4):452\u2013458","journal-title":"Carcinogenesis"},{"issue":"1","key":"482_CR18","doi-asserted-by":"publisher","first-page":"95","DOI":"10.1186\/s13148-023-01511-y","volume":"15","author":"JE Joo","year":"2023","unstructured":"Joo JE et al (2023) Identifying primary and secondary MLH1 epimutation carriers displaying low-level constitutional MLH1 methylation using droplet digital PCR and genome-wide DNA methylation profiling of colorectal cancers. Clin Epigenetics 15(1):95","journal-title":"Clin Epigenetics"},{"issue":"1","key":"482_CR19","doi-asserted-by":"publisher","first-page":"46","DOI":"10.1186\/s13053-021-00203-z","volume":"19","author":"K Wallander","year":"2021","unstructured":"Wallander K et al (2021) Massive parallel sequencing in individuals with multiple primary tumours reveals the benefit of re-analysis. Hered Cancer Clin Pract 19(1):46","journal-title":"Hered Cancer Clin Pract"},{"issue":"1","key":"482_CR20","doi-asserted-by":"publisher","first-page":"23","DOI":"10.1186\/s13053-021-00181-2","volume":"19","author":"K Wallander","year":"2021","unstructured":"Wallander K et al (2021) Massive parallel sequencing in a family with rectal cancer. Hered Cancer Clin Pract 19(1):23","journal-title":"Hered Cancer Clin Pract"},{"issue":"1","key":"482_CR21","doi-asserted-by":"publisher","first-page":"171","DOI":"10.1186\/s13148-019-0762-6","volume":"11","author":"E D\u00e1maso","year":"2019","unstructured":"D\u00e1maso E et al (2019) Highly sensitive MLH1 methylation analysis in blood identifies a cancer patient with low-level mosaic MLH1 epimutation. Clin Epigenetics 11(1):171","journal-title":"Clin Epigenetics"},{"issue":"18","key":"482_CR22","doi-asserted-by":"publisher","first-page":"3094","DOI":"10.1093\/bioinformatics\/bty191","volume":"34","author":"H Li","year":"2018","unstructured":"Li H (2018) Minimap2: pairwise alignment for nucleotide sequences. Bioinformatics 34(18):3094\u20133100","journal-title":"Bioinformatics"},{"issue":"12","key":"482_CR23","doi-asserted-by":"publisher","first-page":"797","DOI":"10.1038\/s43588-022-00387-x","volume":"2","author":"Z Zheng","year":"2022","unstructured":"Zheng Z et al (2022) Symphonizing pileup and full-alignment for deep learning-based long-read variant calling. Nat Comput Sci 2(12):797\u2013803","journal-title":"Nat Comput Sci"},{"issue":"6","key":"482_CR24","doi-asserted-by":"publisher","first-page":"498","DOI":"10.1089\/cmb.2014.0157","volume":"22","author":"M Patterson","year":"2015","unstructured":"Patterson M et al (2015) WhatsHap: weighted haplotype assembly for future-generation sequencing reads. J Comput Biol 22(6):498\u2013509","journal-title":"J Comput Biol"},{"key":"482_CR25","doi-asserted-by":"crossref","unstructured":"Razaghi R et al (2022) Modbamtools: analysis of single-molecule epigenetic data for long-range profiling, heterogeneity, and clustering. bioRxiv, p 2022.07.07.499188","DOI":"10.1101\/2022.07.07.499188"},{"key":"482_CR26","doi-asserted-by":"crossref","unstructured":"Plazzer JP et al (2024) Mismatch repair gene specifications to the ACMG\/AMP classification criteria: consensus recommendations from the InSiGHT ClinGen Hereditary Colorectal Cancer \/ Polyposis Variant Curation Expert Panel. medRxiv, p 2024.05.13.24307108","DOI":"10.1101\/2024.05.13.24307108"},{"issue":"23","key":"482_CR27","doi-asserted-by":"publisher","first-page":"3847","DOI":"10.1093\/bioinformatics\/btv470","volume":"31","author":"SG Coetzee","year":"2015","unstructured":"Coetzee SG, Coetzee GA, Hazelett DJ (2015) motifbreakR: an R\/Bioconductor package for predicting variant effects at transcription factor binding sites. Bioinformatics 31(23):3847\u20133849","journal-title":"Bioinformatics"},{"issue":"4","key":"482_CR28","doi-asserted-by":"publisher","first-page":"869","DOI":"10.1002\/ijc.25422","volume":"128","author":"A Goel","year":"2011","unstructured":"Goel A et al (2011) De novo constitutional MLH1 epimutations confer early-onset colorectal cancer in two new sporadic Lynch syndrome cases, with derivation of the epimutation on the paternal allele in one. Int J Cancer 128(4):869\u2013878","journal-title":"Int J Cancer"},{"issue":"11","key":"482_CR29","doi-asserted-by":"publisher","first-page":"2060","DOI":"10.3390\/genes14112060","volume":"14","author":"IW Carnevali","year":"2023","unstructured":"Carnevali IW et al (2023) MLH1 promoter methylation could be the second hit in lynch syndrome carcinogenesis. Genes (Basel) 14(11):2060","journal-title":"Genes (Basel)"},{"issue":"1","key":"482_CR30","doi-asserted-by":"publisher","first-page":"112","DOI":"10.1038\/ng.283","volume":"41","author":"MJ Ligtenberg","year":"2009","unstructured":"Ligtenberg MJ et al (2009) Heritable somatic methylation and inactivation of MSH2 in families with Lynch syndrome due to deletion of the 3\u2019 exons of TACSTD1. Nat Genet 41(1):112\u2013117","journal-title":"Nat Genet"},{"key":"482_CR31","doi-asserted-by":"publisher","first-page":"26","DOI":"10.1186\/1756-8935-7-26","volume":"7","author":"H Saadeh","year":"2014","unstructured":"Saadeh H, Schulz R (2014) Protection of CpG islands against de novo DNA methylation during oogenesis is associated with the recognition site of E2f1 and E2f2. Epigenetics Chromatin 7:26","journal-title":"Epigenetics Chromatin"}],"container-title":["Familial Cancer"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/link.springer.com\/content\/pdf\/10.1007\/s10689-025-00482-8.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/link.springer.com\/article\/10.1007\/s10689-025-00482-8\/fulltext.html","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/link.springer.com\/content\/pdf\/10.1007\/s10689-025-00482-8.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,9,19]],"date-time":"2025-09-19T04:40:27Z","timestamp":1758256827000},"score":1,"resource":{"primary":{"URL":"https:\/\/link.springer.com\/10.1007\/s10689-025-00482-8"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,7,26]]},"references-count":31,"journal-issue":{"issue":"3","published-online":{"date-parts":[[2025,9]]}},"alternative-id":["482"],"URL":"https:\/\/doi.org\/10.1007\/s10689-025-00482-8","relation":{},"ISSN":["1573-7292"],"issn-type":[{"value":"1573-7292","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,7,26]]},"assertion":[{"value":"7 March 2025","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"15 June 2025","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"26 July 2025","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Declarations"}},{"value":"The authors declare no competing interests.","order":2,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing interests"}},{"value":"This study was performed in line with the principles of the Declaration of Helsinki. The study was approved by the Cedars-Sinai Medical Center Institutional Review Board for research involving human subjects (Hitchins #Pro00049624).","order":3,"name":"Ethics","group":{"name":"EthicsHeading","label":"Ethics approval"}}],"article-number":"59"}}