{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,14]],"date-time":"2026-10-14T11:51:39Z","timestamp":1791978699919,"version":"4.4.0"},"reference-count":35,"publisher":"Wiley","license":[{"start":{"date-parts":[[2026,10,14]],"date-time":"2026-10-14T00:00:00Z","timestamp":1791936000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2026,10,14]],"date-time":"2026-10-14T00:00:00Z","timestamp":1791936000000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"funder":[{"DOI":"10.13039\/100019687","name":"Hamad Bin Khalifa University","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100019687","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100019475","name":"Sidra Medicine","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100019475","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Intl Journal of Cancer"],"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:p>Lynch syndrome, a prevalent hereditary cancer predisposition, is frequently caused by pathogenic variants in the MutL homolog 1 (MLH1) mismatch repair gene. However, a significant proportion of MLH1 missense variants remain classified as variants of uncertain significance (VUS), creating clinical ambiguity. Functional validation assays in vertebrate models are crucial to assess the pathogenicity of these VUS. We previously reported the rare MLH1 NM_000249.4:c.335A&gt;G (p.His112Arg) variant in individuals with cancer histories within the Qatar Genome Program. To assess its functional impact, we established the MLH1 p.His112Arg variant in the human colorectal cancer cell line, COLO205, using CRISPR\u2010Cas9 technology. Isogenic wild\u2010type (WT) and variant\u2010carrying cells were xenotransplanted into the zebrafish. Tumor growth was quantified via fluorescence microscopy at 24 and 72\u2009h post\u2010injection. Zebrafish xenografted with MLH1 p.His112Arg cells exhibited a significant increase in tumor area compared to WT. This accelerated growth was associated with a significant upregulation of the proliferation marker MKI67 in variant\u2010carrying tumors, while cyclin D1 (CCND1) and proliferating cell nuclear antigen (PCNA) (key regulators of cell cycle progression and DNA replication, respectively) expression remained unchanged, despite a tendency for increased expression. Furthermore, our functional characterization of the MLH1 p.His112Arg variant in COLO205 cells showed an enhanced proliferation capacity compared to the WT cell line, as well as increased cell viability and decreased apoptotic susceptibility when subjected to methylation\u2010induced DNA damage. The MLH1 p.His112Arg variant confers a tumor\u2010promoting phenotype in an in\u00a0vivo zebrafish model, providing functional evidence that supports its reclassification from VUS to likely pathogenic.<\/jats:p>","DOI":"10.1002\/ijc.70790","type":"journal-article","created":{"date-parts":[[2026,10,14]],"date-time":"2026-10-14T11:32:14Z","timestamp":1791977534000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["<scp>MLH1<\/scp>\n                    p.\n                    <scp>His112Arg<\/scp>\n                    Variant Is Associated With Increased Cancer Susceptibility"],"prefix":"10.1002","author":[{"given":"Amal","family":"Elfatih","sequence":"first","affiliation":[{"name":"College of Health and Life Science, Genomics and Precision Medicine Hamad Bin Khalifa University  Doha Qatar"},{"name":"Qatar Precision Health Institute  Doha Qatar"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1019-3489","authenticated-orcid":false,"given":"Issam","family":"Tout","sequence":"additional","affiliation":[{"name":"Research Department Sidra Medicine  Doha Qatar"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Najwa Ahmad","family":"Doha","sequence":"additional","affiliation":[{"name":"College of Health and Life Science, Genomics and Precision Medicine Hamad Bin Khalifa University  Doha Qatar"},{"name":"Research Department Sidra Medicine  Doha Qatar"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tala Abu","family":"Arja","sequence":"additional","affiliation":[{"name":"Research Department Sidra Medicine  Doha Qatar"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Idris","family":"Mohammed","sequence":"additional","affiliation":[{"name":"Research Department Sidra Medicine  Doha Qatar"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Doua","family":"Abdelrhaman","sequence":"additional","affiliation":[{"name":"Research Department Sidra Medicine  Doha Qatar"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Waseem","family":"Hasan","sequence":"additional","affiliation":[{"name":"Research Department Sidra Medicine  Doha Qatar"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chadi","family":"Saad","sequence":"additional","affiliation":[{"name":"Qatar Precision Health Institute  Doha Qatar"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hamdi","family":"Mbarek","sequence":"additional","affiliation":[{"name":"Qatar Precision Health Institute  Doha Qatar"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Borbala","family":"Mifsud","sequence":"additional","affiliation":[{"name":"College of Health and Life Science, Genomics and Precision Medicine Hamad Bin Khalifa University  Doha Qatar"},{"name":"William Harvey Research Institute Queen Mary University London  London UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3438-5028","authenticated-orcid":false,"given":"Sahar Isa","family":"Da'as","sequence":"additional","affiliation":[{"name":"College of Health and Life Science, Genomics and Precision Medicine Hamad Bin Khalifa University  Doha Qatar"},{"name":"Research Department Sidra Medicine  Doha Qatar"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2026,10,14]]},"reference":[{"issue":"5","key":"e_1_2_13_2_1","doi-asserted-by":"crossref","first-page":"783","DOI":"10.1053\/j.gastro.2022.08.058","article-title":"Lynch Syndrome Genetics and Clinical Implications","volume":"164","author":"Peltom\u00e4ki P.","year":"2023","journal-title":"Gastroenterology"},{"issue":"9","key":"e_1_2_13_3_1","doi-asserted-by":"crossref","DOI":"10.1002\/hsr2.71286","article-title":"Hereditary Nonpolyposis Colon Cancer (Lynch Syndrome): An Emerging Public Health Concern","volume":"8","author":"Mohiuddin M.","year":"2025","journal-title":"Health Science Reports"},{"issue":"2","key":"e_1_2_13_4_1","doi-asserted-by":"crossref","DOI":"10.3390\/ijms25020850","article-title":"Classification of MLH1 Missense VUS Using Protein Structure\u2010Based Deep Learning\u2010Ramachandran Plot\u2010Molecular Dynamics Simulations Method","volume":"25","author":"Tam B.","year":"2024","journal-title":"International Journal of Molecular Sciences"},{"issue":"11","key":"e_1_2_13_5_1","doi-asserted-by":"crossref","first-page":"1576","DOI":"10.1002\/humu.22142","article-title":"Comprehensive Functional Assessment of MLH1 Variants of Unknown Significance","volume":"33","author":"Borr\u00e0s E.","year":"2012","journal-title":"Human Mutation"},{"issue":"6","key":"e_1_2_13_6_1","doi-asserted-by":"crossref","DOI":"10.1371\/journal.pone.0304141","article-title":"Functional Analysis of MMR Gene VUS From Potential Lynch Syndrome Patients","volume":"19","author":"Mahdouani M.","year":"2024","journal-title":"PLoS One"},{"issue":"12","key":"e_1_2_13_7_1","doi-asserted-by":"crossref","first-page":"2295","DOI":"10.1002\/humu.24462","article-title":"A Calibrated Cell\u2010Based Functional Assay to Aid Classification of MLH1 DNA Mismatch Repair Gene Variants","volume":"43","author":"Rath A.","year":"2022","journal-title":"Human Mutation"},{"issue":"1","key":"e_1_2_13_8_1","doi-asserted-by":"crossref","DOI":"10.1186\/s13073-019-0690-2","article-title":"Recommendations for Application of the Functional Evidence PS3\/BS3 Criterion Using the ACMG\/AMP Sequence Variant Interpretation Framework","volume":"12","author":"Brnich S. E.","year":"2019","journal-title":"Genome Medicine"},{"issue":"24","key":"e_1_2_13_9_1","doi-asserted-by":"crossref","DOI":"10.3390\/ijms222413356","article-title":"Use of Zebrafish Models to Boost Research in Rare Genetic Diseases","volume":"22","author":"Crouzier L.","year":"2021","journal-title":"International Journal of Molecular Sciences"},{"issue":"10","key":"e_1_2_13_10_1","doi-asserted-by":"crossref","first-page":"641","DOI":"10.1136\/jmedgenet-2018-105358","article-title":"Use of Zebrafish Models to Investigate Rare Human Disease","volume":"55","author":"Adamson K. I.","year":"2018","journal-title":"Journal of Medical Genetics"},{"issue":"16","key":"e_1_2_13_11_1","doi-asserted-by":"crossref","first-page":"2796","DOI":"10.1093\/hmg\/ddac073","article-title":"Analysis of Incidental Findings in Qatar Genome Participants Reveals Novel Functional Variants in LMNA and DSP","volume":"31","author":"Elfatih A.","year":"2022","journal-title":"Human Molecular Genetics"},{"key":"e_1_2_13_12_1","doi-asserted-by":"crossref","unstructured":"M.Vanhooydonck S.Debaenst E.Vanbelleghem et\u00a0al. \u201cEvaluating Variants of Uncertain Significance in Adult Knock\u2010In Zebrafish: A Proof of Concept With a COL1A2 Variant \u201d bioRxiv 2025:2025.2002 2011.637642.","DOI":"10.1101\/2025.02.11.637642"},{"issue":"11","key":"e_1_2_13_13_1","doi-asserted-by":"crossref","DOI":"10.3390\/genes10110935","article-title":"Zebrafish Models of Cancer\u2014New Insights on Modeling Human Cancer in a Non\u2010Mammalian Vertebrate","volume":"10","author":"Hason M.","year":"2019","journal-title":"Genes"},{"issue":"11","key":"e_1_2_13_14_1","doi-asserted-by":"crossref","first-page":"1465","DOI":"10.1038\/s41431-024-01656-1","article-title":"Analysis of 14,392 Whole Genomes Reveals 3.5% of Qataris Carry Medically Actionable Variants","volume":"32","author":"Elfatih A.","year":"2024","journal-title":"European Journal of Human Genetics"},{"key":"e_1_2_13_15_1","doi-asserted-by":"crossref","DOI":"10.3389\/fgene.2019.00736","article-title":"Sentieon DNASeq Variant Calling Workflow Demonstrates Strong Computational Performance and Accuracy","volume":"10","author":"Kendig K. 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J.","year":"2016","journal-title":"Nucleic Acids Research"},{"issue":"6","key":"e_1_2_13_19_1","first-page":"577","article-title":"Human Gene Mutation Database (HGMD): 2003 Update","volume":"21","author":"Stenson P. D.","year":"2003","journal-title":"Human Mutation"},{"issue":"9","key":"e_1_2_13_20_1","doi-asserted-by":"crossref","first-page":"1488","DOI":"10.1002\/humu.24051","article-title":"AutoPVS1: An Automatic Classification Tool for PVS1 Interpretation of Null Variants","volume":"41","author":"Xiang J.","year":"2020","journal-title":"Human Mutation"},{"issue":"4","key":"e_1_2_13_21_1","doi-asserted-by":"crossref","first-page":"877","DOI":"10.1016\/j.ajhg.2016.08.016","article-title":"REVEL: An Ensemble Method for Predicting the Pathogenicity of Rare Missense Variants","volume":"99","author":"Ioannidis N. 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E.Ashour E.Krekeler M. C.Bhowmik et\u00a0al. \u201cHuman MutL\u03b1 Activates Methylpurine DNA Glycosylase to Induce Alkylation Damage Cytotoxicity \u201d bioRxiv 2025.","DOI":"10.64898\/2025.12.15.694403"}],"container-title":["International Journal of Cancer"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/ijc.70790","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/full-xml\/10.1002\/ijc.70790","content-type":"application\/xml","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/ijc.70790","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,10,14]],"date-time":"2026-10-14T11:32:21Z","timestamp":1791977541000},"score":1,"resource":{"primary":{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/10.1002\/ijc.70790"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,10,14]]},"references-count":35,"alternative-id":["10.1002\/ijc.70790"],"URL":"https:\/\/doi.org\/10.1002\/ijc.70790","archive":["Portico"],"relation":{},"ISSN":["0020-7136","1097-0215"],"issn-type":[{"value":"0020-7136","type":"print"},{"value":"1097-0215","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,10,14]]},"assertion":[{"value":"2026-05-06","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2026-09-30","order":2,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2026-10-14","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}],"article-number":"ijc.70790"}}