{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2023,9,20]],"date-time":"2023-09-20T19:06:49Z","timestamp":1695236809580},"reference-count":43,"publisher":"Wiley","issue":"3","license":[{"start":{"date-parts":[[2015,2,24]],"date-time":"2015-02-24T00:00:00Z","timestamp":1424736000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Haemophilia"],"published-print":{"date-parts":[[2015,5]]},"abstract":"<jats:title>Summary<\/jats:title><jats:p>This study aims to determine the way to predict the haemophilia A (<jats:styled-content style=\"fixed-case\">HA<\/jats:styled-content>) carrier status and the potential severity in six females with low FVIII:C levels (&lt;0.50\u00a0IU\u00a0mL<jats:sup>\u22121<\/jats:sup>), <jats:italic>F8<\/jats:italic> gene variations and without family history of <jats:styled-content style=\"fixed-case\">HA<\/jats:styled-content>. Except p.Ser577Tyr, <jats:italic>F8<\/jats:italic> gene variations that we reported have never been described (p.Leu107His, p.Pro521Leu, p.Val682Leu, p.Leu2032Pro, p.Ala315dup). Prediction of their potential causal impact was studied by two strategies: bioinformatics approaches and site\u2010directed mutagenesis followed by FVIII cellular expression into <jats:styled-content style=\"fixed-case\">COS<\/jats:styled-content>\u20101 cell. FVIII clotting assay (FVIII:C) and antigen (FVIII:Ag) were assayed <jats:italic>in vitro<\/jats:italic>. <jats:italic>In silico<\/jats:italic> analysis showed the probably damaging effect of all substitutions and the full conservation of the residues across mammalian species, except for p.Leu2032Pro. The <jats:italic>in vitro<\/jats:italic> variant expression model showed abnormal intra and\/or extracellular FVIII:C and FVIII:Ag levels for five mutations, which suggest their causality in <jats:styled-content style=\"fixed-case\">HA<\/jats:styled-content> and provide informations about the involved mechanism. We suspect a defect in synthesis and secretion for p.Leu107His, p.Ala315dup and p.Pro521Leu. The mutation p.Val682Leu only affects the FVIII function while p.Ser577Tyr alters function and synthesis. The variant p.Leu2032Pro is probably a polymorphism because no alteration of the FVIII protein expression was observed <jats:italic>in vitro<\/jats:italic>. <jats:italic>In vitro<\/jats:italic> results suggest that mutations p.Ser577Tyr and p.Ala315dup could led to a severe <jats:styled-content style=\"fixed-case\">HA<\/jats:styled-content> in men. This study demonstrates the ability of this <jats:italic>in vitro<\/jats:italic> cellular expression model to contribute to the diagnosis strategy for female suspected of being <jats:styled-content style=\"fixed-case\">HA<\/jats:styled-content> carrier, without <jats:styled-content style=\"fixed-case\">HA<\/jats:styled-content> family history and with a novel <jats:italic>F8<\/jats:italic> gene variation and to provide new criteria for the genetic counselling.<\/jats:p>","DOI":"10.1111\/hae.12651","type":"journal-article","created":{"date-parts":[[2015,2,24]],"date-time":"2015-02-24T06:25:22Z","timestamp":1424759122000},"source":"Crossref","is-referenced-by-count":7,"title":["Usefulness of an <i>in vitro<\/i> cellular expression model for haemophilia A carrier diagnosis: illustration with five novel mutations in the <i>F8<\/i> gene in women with isolated factor VIII:C deficiency"],"prefix":"10.1111","volume":"21","author":[{"given":"O.","family":"Roualdes","sequence":"first","affiliation":[{"name":"Hospices Civils de Lyon H\u00f4pital Edouard Herriot Service d'H\u00e9matologie Biologique Lyon"},{"name":"EAM 4174 H\u00e9mostase Inflammation et Sepsis Universit\u00e9 Claude Bernard Lyon 1 Lyon"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"C.","family":"Nougier","sequence":"additional","affiliation":[{"name":"Hospices Civils de Lyon H\u00f4pital Edouard Herriot Service d'H\u00e9matologie Biologique Lyon"},{"name":"EAM 4174 H\u00e9mostase Inflammation et Sepsis Universit\u00e9 Claude Bernard Lyon 1 Lyon"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"M.","family":"Fretigny","sequence":"additional","affiliation":[{"name":"Hospices Civils de Lyon H\u00f4pital Edouard Herriot Service d'H\u00e9matologie Biologique Lyon"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"E.","family":"Talagrand","sequence":"additional","affiliation":[{"name":"Hospices Civils de Lyon H\u00f4pital Edouard Herriot Service d'H\u00e9matologie Biologique Lyon"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"B.","family":"Durand","sequence":"additional","affiliation":[{"name":"Hospices Civils de Lyon H\u00f4pital de la Croix Rousse Laboratoire d'H\u00e9matologie Biologique Lyon"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"C.","family":"Negrier","sequence":"additional","affiliation":[{"name":"Hospices Civils de Lyon H\u00f4pital Edouard Herriot Service d'H\u00e9matologie Biologique Lyon"},{"name":"EAM 4174 H\u00e9mostase Inflammation et Sepsis Universit\u00e9 Claude Bernard Lyon 1 Lyon"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"C.","family":"Vinciguerra","sequence":"additional","affiliation":[{"name":"Hospices Civils de Lyon H\u00f4pital Edouard Herriot Service d'H\u00e9matologie Biologique Lyon"},{"name":"EAM 4174 H\u00e9mostase Inflammation et Sepsis Universit\u00e9 Claude Bernard Lyon 1 Lyon"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2015,2,24]]},"reference":[{"key":"e_1_2_7_2_1","doi-asserted-by":"publisher","DOI":"10.1055\/s-0037-1615621"},{"key":"e_1_2_7_3_1","doi-asserted-by":"publisher","DOI":"10.1055\/s-0038-1647493"},{"key":"e_1_2_7_4_1","doi-asserted-by":"publisher","DOI":"10.1016\/0049-3848(82)90131-1"},{"key":"e_1_2_7_5_1","doi-asserted-by":"publisher","DOI":"10.1182\/blood-2005-09-3879"},{"key":"e_1_2_7_6_1","doi-asserted-by":"publisher","DOI":"10.1016\/0049-3848(82)90130-X"},{"key":"e_1_2_7_7_1","first-page":"135","article-title":"Sex chromatin and gene action in the mammalian X\u2010chromosome","volume":"14","author":"Lyon MF","year":"1962","journal-title":"Am J Hum Genet"},{"key":"e_1_2_7_8_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1365-2141.2004.05275.x"},{"key":"e_1_2_7_9_1","doi-asserted-by":"publisher","DOI":"10.1182\/blood-2002-07-2081"},{"key":"e_1_2_7_10_1","doi-asserted-by":"publisher","DOI":"10.1016\/S1050-1738(00)00036-0"},{"key":"e_1_2_7_11_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.274.53.37685"},{"key":"e_1_2_7_12_1","first-page":"89","article-title":"Factor VIII and factor IX in a twin population. Evidence for a major effect of ABO locus on factor VIII level","volume":"37","author":"Orstavik KH","year":"1985","journal-title":"Am J Hum Genet"},{"key":"e_1_2_7_13_1","doi-asserted-by":"publisher","DOI":"10.1046\/j.1538-7836.2003.00314.x"},{"key":"e_1_2_7_14_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1365-2141.1981.00651.x"},{"key":"e_1_2_7_15_1","doi-asserted-by":"publisher","DOI":"10.1038\/ng1193-236"},{"key":"e_1_2_7_16_1","doi-asserted-by":"publisher","DOI":"10.1182\/blood.V99.1.168"},{"key":"e_1_2_7_17_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1538-7836.2006.01840.x"},{"key":"e_1_2_7_18_1","doi-asserted-by":"publisher","DOI":"10.1002\/(SICI)1098-1004(1998)11:1<1::AID-HUMU1>3.0.CO;2-O"},{"key":"e_1_2_7_19_1","doi-asserted-by":"publisher","DOI":"10.1002\/humu.10262"},{"key":"e_1_2_7_20_1","doi-asserted-by":"publisher","DOI":"10.1055\/s-0037-1616092"},{"key":"e_1_2_7_21_1","unstructured":"PolyPhen\u20102: prediction of functional effects of human nsSNPs. Available athttp:\/\/genetics.bwh.harvard.edu\/pph2\/. Accessed October 10 2013."},{"key":"e_1_2_7_22_1","doi-asserted-by":"publisher","DOI":"10.1038\/nmeth0410-248"},{"key":"e_1_2_7_23_1","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gks539"},{"key":"e_1_2_7_24_1","unstructured":"SIFT (Sorting Intolerant From Tolerant) \u2013 Tool to predict nonsynonmous\/missense variants. Available athttp:\/\/sift.bii.a-star.edu.sg\/. Accessed October 10 2013"},{"key":"e_1_2_7_25_1","unstructured":"International Agency for Research on Cancer (IRAC).Align GVGD. Available athttp:\/\/agvgd.iarc.fr\/. Accessed October 10 2013"},{"key":"e_1_2_7_26_1","doi-asserted-by":"publisher","DOI":"10.1002\/humu.21490"},{"key":"e_1_2_7_27_1","first-page":"115","article-title":"Novel human pathological mutations. Gene symbol: F8. Disease: haemophilia A","volume":"127","author":"Niceta M","year":"2010","journal-title":"Hum Genet"},{"key":"e_1_2_7_28_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.270.17.10297"},{"key":"e_1_2_7_29_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.273.14.8537"},{"key":"e_1_2_7_30_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.272.39.24121"},{"key":"e_1_2_7_31_1","doi-asserted-by":"publisher","DOI":"10.1021\/bi0520380"},{"key":"e_1_2_7_32_1","doi-asserted-by":"publisher","DOI":"10.1002\/humu.20345"},{"key":"e_1_2_7_33_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0021-9258(17)32024-0"},{"key":"e_1_2_7_34_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M008046200"},{"key":"e_1_2_7_35_1","doi-asserted-by":"publisher","DOI":"10.1182\/blood.V91.2.538"},{"key":"e_1_2_7_36_1","doi-asserted-by":"publisher","DOI":"10.1182\/blood-2001-12-0361"},{"key":"e_1_2_7_37_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.274.34.23734"},{"key":"e_1_2_7_38_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1365-2516.2011.02700.x"},{"key":"e_1_2_7_39_1","doi-asserted-by":"publisher","DOI":"10.1006\/geno.1993.1073"},{"key":"e_1_2_7_40_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1538-7836.2010.04174.x"},{"key":"e_1_2_7_41_1","doi-asserted-by":"publisher","DOI":"10.1182\/blood.V92.11.3983"},{"key":"e_1_2_7_42_1","doi-asserted-by":"publisher","DOI":"10.1055\/s-0037-1615834"},{"key":"e_1_2_7_43_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0168-1656(98)00035-2"},{"key":"e_1_2_7_44_1","doi-asserted-by":"publisher","DOI":"10.1182\/blood-2003-10-3591"}],"container-title":["Haemophilia"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/api.wiley.com\/onlinelibrary\/tdm\/v1\/articles\/10.1111%2Fhae.12651","content-type":"unspecified","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1111\/hae.12651","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/full-xml\/10.1111\/hae.12651","content-type":"application\/xml","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1111\/hae.12651","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,8,24]],"date-time":"2023-08-24T12:31:55Z","timestamp":1692880315000},"score":1,"resource":{"primary":{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/10.1111\/hae.12651"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2015,2,24]]},"references-count":43,"journal-issue":{"issue":"3","published-print":{"date-parts":[[2015,5]]}},"alternative-id":["10.1111\/hae.12651"],"URL":"https:\/\/doi.org\/10.1111\/hae.12651","archive":["Portico"],"relation":{},"ISSN":["1351-8216","1365-2516"],"issn-type":[{"value":"1351-8216","type":"print"},{"value":"1365-2516","type":"electronic"}],"subject":[],"published":{"date-parts":[[2015,2,24]]}}}