{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,30]],"date-time":"2026-09-30T00:54:12Z","timestamp":1790729652973,"version":"4.1.0"},"reference-count":30,"publisher":"Wiley","issue":"5","license":[{"start":{"date-parts":[[2023,7,6]],"date-time":"2023-07-06T00:00:00Z","timestamp":1688601600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Haemophilia"],"published-print":{"date-parts":[[2023,9]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:sec><jats:title>Introduction<\/jats:title><jats:p>Conventional genetic investigation fails to identify the <jats:italic>F8<\/jats:italic> causal variant in 2.5%\u201010% of haemophilia A (HA) patients with non\u2010severe phenotypes. In these cases, <jats:italic>F8<\/jats:italic> deep intronic variants could be causal.<\/jats:p><\/jats:sec><jats:sec><jats:title>Aim<\/jats:title><jats:p>To identify pathogenic <jats:italic>F8<\/jats:italic> deep intronic variants in genetically unresolved families with non\u2010severe HA analysed in the haematology laboratory of the Hospices Civils de Lyon.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>The whole <jats:italic>F8<\/jats:italic> was analysed by next generation sequencing. The pathogenic impact of candidate variants identified was assessed using both in silico analysis (MaxEntScan and spliceAI) and functional analysis (RNA or minigene assay).<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>Sequencing was performed in 49\/55 families included for which a DNA sample from a male propositus was available. In total, 33 candidate variants from 43 propositi were identified. These variants corresponded to 31 single nucleotide substitutions, one 173\u2010bp deletion, and an 869\u2010bp tandem triplication. No candidate variant was found in six propositi. The most frequent variants found were the association of [c.2113+1154G&gt;C and c.5374\u2010304C&gt;T], identified in five propositi, and the c.2114\u20106529C&gt;G identified in nine propositi. Four variants had been previously described as HA\u2010causing. Splicing functional assay found a deleterious impact for 11 substitutions (c.671\u201094G&gt;A, c.788\u2010312A&gt;G, c.2113+1154G&gt;C, c.2114\u20106529C&gt;G, c.5999\u2010820A&gt;T, c.5999\u2010786C&gt;A, c.5999\u2010669G&gt;T, c.5999\u2010669G&gt;A, c.5999\u2010669G&gt;C, c.6900+4104A&gt;C, and c.6901\u20102992A&gt;G). The HA\u2010causing variant was identified in 33\/49 (67%) cases. In total, <jats:italic>F8<\/jats:italic> deep intronic variants caused 8.8% of the non\u2010severe HA among the 1643 families analysed in our laboratory.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusion<\/jats:title><jats:p>The results emphasise the value of whole <jats:italic>F8<\/jats:italic> gene sequencing combined with splicing functional analyses to improve the diagnosis yield for non\u2010severe HA.<\/jats:p><\/jats:sec>","DOI":"10.1111\/hae.14824","type":"journal-article","created":{"date-parts":[[2023,7,6]],"date-time":"2023-07-06T17:44:41Z","timestamp":1688665481000},"page":"1320-1333","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":7,"title":["Whole F8 gene sequencing combined with splicing functional analyses led to a substantial increase of the molecular diagnosis yield for non\u2010severe haemophilia A"],"prefix":"10.1111","volume":"29","author":[{"given":"Amy","family":"Dericquebourg","sequence":"first","affiliation":[{"name":"Hospices Civils de Lyon Groupe Hospitalier Est Service d'h\u00e9matologie biologique Bron France"},{"name":"Universit\u00e9 Claude Bernard Lyon 1 UR4609 H\u00e9mostase et thrombose Lyon France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mathilde","family":"Fretigny","sequence":"additional","affiliation":[{"name":"Hospices Civils de Lyon Groupe Hospitalier Est Service d'h\u00e9matologie biologique Bron France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alexandre","family":"Leuci","sequence":"additional","affiliation":[{"name":"Universit\u00e9 Claude Bernard Lyon 1 UR4609 H\u00e9mostase et thrombose Lyon France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Christophe","family":"Zawadzki","sequence":"additional","affiliation":[{"name":"P\u00f4le de Biologie Pathologie G\u00e9n\u00e9tique Institut d'H\u00e9matologie \u2010 Transfusion CHU Lille Lille France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yoann","family":"Huguenin","sequence":"additional","affiliation":[{"name":"Centre de Ressources et de Comp\u00e9tence des Maladies H\u00e9morragiques Constitutionnelles H\u00f4pital Pellegrin, CHU de Bordeaux Bordeaux France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sabine\u2010Marie","family":"Castet","sequence":"additional","affiliation":[{"name":"Centre de Ressources et de Comp\u00e9tence des Maladies H\u00e9morragiques Constitutionnelles H\u00f4pital Pellegrin, CHU de Bordeaux Bordeaux France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yesim","family":"Dargaud","sequence":"additional","affiliation":[{"name":"Universit\u00e9 Claude Bernard Lyon 1 UR4609 H\u00e9mostase et thrombose Lyon France"},{"name":"Unit\u00e9 d'H\u00e9mostase Clinique Centre National de Reference de l'H\u00e9mophilie H\u00f4pital Cardiologique Louis Pradel Universit\u00e9 Lyon Lyon France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Christine","family":"Vinciguerra","sequence":"additional","affiliation":[{"name":"Hospices Civils de Lyon Groupe Hospitalier Est Service d'h\u00e9matologie biologique Bron France"},{"name":"Universit\u00e9 Claude Bernard Lyon 1 UR4609 H\u00e9mostase et thrombose Lyon France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7408-6474","authenticated-orcid":false,"given":"Yohann","family":"Jourdy","sequence":"additional","affiliation":[{"name":"Hospices Civils de Lyon Groupe Hospitalier Est Service d'h\u00e9matologie biologique Bron France"},{"name":"Universit\u00e9 Claude Bernard Lyon 1 UR4609 H\u00e9mostase et thrombose Lyon 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