{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T13:04:38Z","timestamp":1785416678016,"version":"3.56.0"},"posted":{"date-parts":[[2025,11,14]]},"group-title":"Cell Biology","reference-count":49,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2025,11,14]],"date-time":"2025-11-14T00:00:00Z","timestamp":1763078400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Stand Up To Cancer-Fanconi Cancer Foundation-Farrah Fawcett Foundation Head and Neck Cancer Research Team Grant"},{"DOI":"10.13039\/100002086","name":"Fanconi Cancer Foundation","doi-asserted-by":"crossref","id":[{"id":"10.13039\/100002086","id-type":"DOI","asserted-by":"crossref"}]},{"award":["T2019-013"],"award-info":[{"award-number":["T2019-013"]}],"id":[{"id":"https:\/\/ror.org\/00kbbk236","id-type":"ROR","asserted-by":"publisher"}]},{"award":["GM140400"],"award-info":[{"award-number":["GM140400"]}],"id":[{"id":"https:\/\/ror.org\/01cwqze88","id-type":"ROR","asserted-by":"publisher"}]},{"award":["F30CA268717"],"award-info":[{"award-number":["F30CA268717"]}],"id":[{"id":"https:\/\/ror.org\/01cwqze88","id-type":"ROR","asserted-by":"publisher"}]},{"award":["T32GM152349"],"award-info":[{"award-number":["T32GM152349"]}],"id":[{"id":"https:\/\/ror.org\/01cwqze88","id-type":"ROR","asserted-by":"publisher"}]},{"name":"The Rockefeller University Anderson Center for Cancer Research"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2025,11,14]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:p>The Fanconi anemia (FA) pathway is vital for the repair of DNA interstrand crosslinks (ICLs), which are caused by a variety of endogenous and exogenous genotoxins including reactive aldehydes. Patients with pathogenic variants in the FA pathway are predisposed to early-onset, aggressive malignancies, especially leukemia and head and neck, esophageal, and anogenital squamous cell carcinomas (SCCs). Prior studies have linked endogenous formaldehyde and acetaldehyde in hematopoietic stem cells with bone marrow failure and leukemia in FA-deficient mice. However, the genotoxic aldehydes specific to mucosal keratinocytes, precursors of FA-associated SCCs, remain to be identified.<\/jats:p>\n                <jats:p>\n                  Here, to identify alcohol dehydrogenases (ADHs) and aldehyde dehydrogenases (ALDHs) necessary for protection of keratinocyte genomes from endogenous metabolites, we used a sensitized background of FA-pathway deficiency. We systematically inactivated all highly expressed ADH and ALDHs genes in\n                  <jats:italic>FANCA<\/jats:italic>\n                  -deficient keratinocyte cell lines and identified genes required for their survival. We report that loss of ADH5 or ALDH3-family enzymes in FA pathway-deficient cells has a synthetic lethal effect and induces DNA damage markers, nominating these genes as important defense mechanisms in the detoxification of endogenous aldehydes and prevention of carcinogenesis in keratinocytes. Loss of ADH5 increases formaldehyde levels, while simultaneous loss of four functionally redundant ALDH3 isozymes is predicted to cause accumulation of lipid aldehydes. FA-deficient keratinocytes are more sensitive to treatment with formaldehyde and the lipid aldehyde 4-hydroxynonenal (4HNE) compared to FA-competent cells. In addition, the thiol-rich antioxidant N-acetyl-L-cysteine (NAC) partially rescued the growth of\n                  <jats:italic>\n                    FANCA\n                    <jats:sup>\u2212\/\u2212<\/jats:sup>\n                    ADH5\n                    <jats:sup>\u2212\/\u2212<\/jats:sup>\n                  <\/jats:italic>\n                  cells and lowered the level of endogenous DNA damage. This work identifies important defense mechanisms in keratinocytes and suggests cancer preventive strategies in patients with Fanconi anemia.\n                <\/jats:p>\n                <jats:sec>\n                  <jats:title>Significance<\/jats:title>\n                  <jats:p>This work shows that ADH5 and ALDH3-family enzymes function as tier 1 defense mechanisms against endogenous DNA-damaging metabolites in keratinocytes. Enhanced detoxification of endogenous aldehydes might prevent DNA damage and mutagenesis, which drive Fanconi anemia-associated head and neck and other mucosal squamous cell carcinomas.<\/jats:p>\n                <\/jats:sec>","DOI":"10.1101\/2025.11.13.688345","type":"posted-content","created":{"date-parts":[[2025,11,14]],"date-time":"2025-11-14T22:40:15Z","timestamp":1763160015000},"source":"Crossref","is-referenced-by-count":3,"title":["Genotoxic formaldehyde and lipid aldehydes are sources of DNA damage in keratinocytes"],"prefix":"10.64898","author":[{"given":"Nicolas J.","family":"Blobel","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yibing","family":"Yao","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marian C.","family":"Okondo","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6285-1562","authenticated-orcid":false,"given":"Agata","family":"Smogorzewska","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"54368","reference":[{"key":"2025111804350886000_2025.11.13.688345v1.1","doi-asserted-by":"publisher","DOI":"10.1038\/nature11863"},{"key":"2025111804350886000_2025.11.13.688345v1.2","doi-asserted-by":"publisher","DOI":"10.3324\/haematol.2017.178111"},{"key":"2025111804350886000_2025.11.13.688345v1.3","doi-asserted-by":"crossref","unstructured":"B. A. Harrison et al., Deficiency of the Fanconi anemia core complex protein FAAP100 results in severe Fanconi anemia. J Clin Invest 135, (2025).","DOI":"10.1172\/JCI185126"},{"key":"2025111804350886000_2025.11.13.688345v1.4","doi-asserted-by":"crossref","unstructured":"J. Kuehl et al., Genetic inactivation of FAAP100 causes Fanconi anemia due to disruption of the monoubiquitin ligase core complex. J Clin Invest 135, (2025).","DOI":"10.1172\/JCI187323"},{"key":"2025111804350886000_2025.11.13.688345v1.5","doi-asserted-by":"crossref","first-page":"272","DOI":"10.1002\/ajh.27520","article-title":"Prognostic significance of mutation type and chromosome fragility in Fanconi anemia","volume":"100","year":"2025","journal-title":"Am J Hematol"},{"key":"2025111804350886000_2025.11.13.688345v1.6","doi-asserted-by":"publisher","DOI":"10.3324\/haematol.2015.133520"},{"key":"2025111804350886000_2025.11.13.688345v1.7","unstructured":"A. Smogorzewska , in Fanconi Anemia Clinical Care Guidelines, I. 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Oka et al., Digenic mutations in ALDH2 and ADH5 impair formaldehyde clearance and cause a multisystem disorder, AMeD syndrome. Sci Adv 6, (2020).","DOI":"10.1126\/sciadv.abd7197"},{"key":"2025111804350886000_2025.11.13.688345v1.19","doi-asserted-by":"publisher","DOI":"10.1016\/j.molcel.2015.08.020"},{"key":"2025111804350886000_2025.11.13.688345v1.20","doi-asserted-by":"crossref","unstructured":"M. Jung et al., ALDH9A1 deficiency as a source of endogenous DNA damage that requires repair by the Fanconi anemia pathway. J Cell Biol 224, (2025).","DOI":"10.1083\/jcb.202407141"},{"key":"2025111804350886000_2025.11.13.688345v1.21","doi-asserted-by":"publisher","DOI":"10.1016\/j.molcel.2010.06.023"},{"key":"2025111804350886000_2025.11.13.688345v1.22","doi-asserted-by":"crossref","unstructured":"V. Xanthis , T. Mantso , A. Dimtsi , A. Pappa , V. E. Fadouloglou , Human Aldehyde Dehydrogenases: A Superfamily of Similar Yet Different Proteins Highly Related to Cancer. 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