{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T01:18:19Z","timestamp":1760145499004,"version":"build-2065373602"},"reference-count":43,"publisher":"MDPI AG","issue":"15","license":[{"start":{"date-parts":[[2024,7,31]],"date-time":"2024-07-31T00:00:00Z","timestamp":1722384000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Romanian Society of Hematology"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Hemophilia A (HA) is an X-linked recessive bleeding disorder caused by mutations in the F8 gene, resulting in deficient or dysfunctional factor VIII (FVIII). This study aimed to characterize the mutational profile of HA in Romanian patients using next-generation sequencing (NGS) and multiplex ligation-dependent probe amplification (MLPA). A total of 107 patients were analyzed, revealing pathogenic or likely pathogenic variants in 96.3% of cases. The identified mutations included missense (30.5%), nonsense (9.1%), small deletions (6.4%), small insertions (2.1%), splice-site variants (4.3%), large deletions (1.6%), and large duplications (1.1%). Large intron inversion was previously found in 37.5% of the patients. Novel variants accounted for 21.5% of identified mutations, expanding the spectrum of F8 variants in this population. This study underscores the genetic heterogeneity of HA and provides insights into genotype\u2013phenotype correlations, aiding in clinical management and prenatal diagnosis.<\/jats:p>","DOI":"10.3390\/ijms25158366","type":"journal-article","created":{"date-parts":[[2024,8,1]],"date-time":"2024-08-01T12:38:20Z","timestamp":1722515900000},"page":"8366","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Mutational Profile in Romanian Patients with Hemophilia A"],"prefix":"10.3390","volume":"25","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-3422-5748","authenticated-orcid":false,"given":"Andra","family":"Grigore","sequence":"first","affiliation":[{"name":"Hematology (Clinic and Laboratory) Discipline-Fundeni Clinical Institute, \u201cCarol Davila\u201d University of Medicine and Pharmacy, 020021 Bucharest, Romania"},{"name":"Department of Hematology and Bone Marrow Transplant, Fundeni Clinical Institute, 022328 Bucharest, Romania"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8529-1107","authenticated-orcid":false,"given":"Mihaela","family":"Dragomir","sequence":"additional","affiliation":[{"name":"Department of Hematology and Bone Marrow Transplant, Fundeni Clinical Institute, 022328 Bucharest, Romania"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Onda-Tabita","family":"C\u0103lug\u0103ru","sequence":"additional","affiliation":[{"name":"Department of Hematology and Bone Marrow Transplant, Fundeni Clinical Institute, 022328 Bucharest, Romania"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Dumitru","family":"Jardan","sequence":"additional","affiliation":[{"name":"Molecular Biology Laboratory, Medlife, 010093 Bucharest, Romania"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Cerasela","family":"Jardan","sequence":"additional","affiliation":[{"name":"Department of Hematology and Bone Marrow Transplant, Fundeni Clinical Institute, 022328 Bucharest, Romania"},{"name":"Pediatrics Discipline-Fundeni Clinical Institute, \u201cCarol Davila\u201d University of Medicine and Pharmacy, 020021 Bucharest, Romania"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Melen","family":"Br\u00eenz\u0103","sequence":"additional","affiliation":[{"name":"Department of Hematology and Bone Marrow Transplant, Fundeni Clinical Institute, 022328 Bucharest, Romania"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Paul","family":"B\u0103l\u0103nescu","sequence":"additional","affiliation":[{"name":"Internal Medicine Discipline-Colentina Clinical Hospital, \u201cCarol Davila\u201d University of Medicine and Pharmacy, 020021 Bucharest, Romania"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Daniel","family":"Coriu","sequence":"additional","affiliation":[{"name":"Hematology (Clinic and Laboratory) Discipline-Fundeni Clinical Institute, \u201cCarol Davila\u201d University of Medicine and Pharmacy, 020021 Bucharest, Romania"},{"name":"Department of Hematology and Bone Marrow Transplant, Fundeni Clinical Institute, 022328 Bucharest, Romania"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2024,7,31]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1773","DOI":"10.1056\/NEJM200106073442307","article-title":"The Hemophilias\u2014From royal genes to gene therapy","volume":"344","author":"Mannucci","year":"2001","journal-title":"N. Engl. J. Med."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"8537","DOI":"10.1074\/jbc.273.14.8537","article-title":"Differential interaction of coagulation factor VIII and factor V with protein chaperones calnexin and calreticulin","volume":"273","author":"Pipe","year":"1998","journal-title":"J. Biol. Chem."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"36","DOI":"10.1111\/jth.12412","article-title":"Human liver sinusoidal endothelial cells but not hepatocytes contain factor VIII","volume":"12","author":"Shahani","year":"2014","journal-title":"J. Thromb. Haemost."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"337","DOI":"10.1038\/312337a0","article-title":"Structure of human factor VIII","volume":"312","author":"Vehar","year":"1984","journal-title":"Nature"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"10451","DOI":"10.1016\/S0021-9258(19)81537-5","article-title":"Association of the factor VIII light chain with von Willebrand factor","volume":"263","author":"Lollar","year":"1988","journal-title":"J. Biol. Chem."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"6352","DOI":"10.1016\/S0021-9258(18)68793-9","article-title":"Synthesis, processing, and secretion of recombinant human factor VIII expressed in mammalian cells","volume":"263","author":"Kaufman","year":"1988","journal-title":"J. Biol. Chem."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"505","DOI":"10.1021\/bi00350a035","article-title":"Proteolytic processing of human factor VIII. Correlation of specific cleavages by thrombin, factor Xa, and activated protein C with activation and inactivation of factor VIII coagulant activity","volume":"25","author":"Eaton","year":"1986","journal-title":"Biochemistry"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"326","DOI":"10.1038\/312326a0","article-title":"Characterization of the human factor VIII gene","volume":"312","author":"Gitschier","year":"1984","journal-title":"Nature"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"2206","DOI":"10.1182\/blood.V86.6.2206.bloodjournal8662206","article-title":"Factor VIII gene inversions in severe hemophilia A: Results of an international consortium study","volume":"86","author":"Antonarakis","year":"1995","journal-title":"Blood"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"133","DOI":"10.1111\/j.1365-2516.2004.01005.x","article-title":"Molecular basis of haemophilia A","volume":"10","author":"Oldenburg","year":"2004","journal-title":"Haemophilia"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1001","DOI":"10.1086\/302065","article-title":"Methylation levels at selected CpG Sites in the factor VIII and FGFR3 genes, in mature female and male germ cells: Implications for male-driven evolution","volume":"63","author":"Olek","year":"1998","journal-title":"Am. J. Hum. Genet."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"228","DOI":"10.1007\/s004390050810","article-title":"Variation of site-specific methylation patterns in the factor VIII (F8C) gene in human sperm DNA","volume":"103","author":"Millar","year":"1998","journal-title":"Hum. Genet."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"216","DOI":"10.1093\/nar\/26.1.216","article-title":"The factor VIII structure and mutation resource site: HAMSTeRS Version 4","volume":"26","author":"Tuddenham","year":"1998","journal-title":"Nucleic Acids Res."},{"key":"ref_14","unstructured":"(2024, June 12). EAHAD Factor VIII Variant Database. Available online: https:\/\/f8-db.eahad.org\/."},{"key":"ref_15","unstructured":"Carcao, M., and Goudemand, J. (2018). Inhibitors in Hemophilia: A Primer, World Federation of Hemophilia. [5th ed.]."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"382","DOI":"10.1055\/s-2007-1000659","article-title":"Factor VIII inhibitors in patients with hemophilia A: Epidemiology of inhibitor development and induction of immune tolerance for factor VIII","volume":"21","author":"Kreuz","year":"1995","journal-title":"Semin. Thromb. Hemost"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"82","DOI":"10.1053\/j.seminhematol.2003.11.016","article-title":"Environmental and genetic factors influencing inhibitor development","volume":"41","author":"Oldenburg","year":"2004","journal-title":"Semin. Hematol."},{"key":"ref_18","unstructured":"World Federation of Hemophilia (2023). World Federation of Hemophilia Report on the Annual Global Survey 2022, World Federation of Hemophilia."},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"Brinza, M., Grigore, A., Dragomir, M., Jardan, D., Jardan, C., Balanescu, P., Tarniceriu, C.C., Badulescu, O.V., Blag, C., and Tomuleasa, C. (2023). Large intron inversions in Romanian patients with hemophilia A\u2014First Report. Medicina, 59.","DOI":"10.3390\/medicina59101821"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"96","DOI":"10.1532\/IJH97.06012","article-title":"New Insight into the Molecular Basis of Hemophilia A","volume":"83","author":"Oldenburg","year":"2006","journal-title":"Int. J. Hematol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"S25","DOI":"10.5482\/HAMO-15-05-0017","article-title":"Mutational Spectrum and Deep Intronic Variants in the Factor VIII Gene of Haemophilia a Patients. Identification by next Generation Sequencing","volume":"36","author":"Bach","year":"2016","journal-title":"Hamostaseologie"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"2922","DOI":"10.1182\/blood-2011-09-379453","article-title":"F8 Gene Mutation Type and Inhibitor Development in Patients with Severe Hemophilia A: Systematic Review and Meta-Analysis","volume":"119","author":"Gouw","year":"2012","journal-title":"Blood"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"824","DOI":"10.1182\/bloodadvances.2016002923","article-title":"Novel approach to genetic analysis and results in 3000 hemophilia patients enrolled in the My Life, Our Future initiative","volume":"1","author":"Johnsen","year":"2017","journal-title":"Blood Adv."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"15","DOI":"10.1111\/j.1365-2516.2006.01361.x","article-title":"Genetic risk factors for inhibitors to factors VIII and IX","volume":"12","author":"Oldenburg","year":"2006","journal-title":"Haemophilia"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"488","DOI":"10.1038\/nrg1617","article-title":"Haemophilia A: From mutation analysis to new therapies","volume":"6","author":"Graw","year":"2005","journal-title":"Nat. Rev. Genet."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"11","DOI":"10.1055\/s-2003-37935","article-title":"Structure and function of the factor VIII gene and protein","volume":"29","author":"Thompson","year":"2003","journal-title":"Semin. Thromb. Hemost."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"1183","DOI":"10.1111\/j.1538-7836.2011.04268.x","article-title":"Most factor VIII B domain missense mutations are unlikely to be causative mutations for severe hemophilia A: Implications for genotyping","volume":"9","author":"Ogata","year":"2011","journal-title":"J. Thromb. Haemost."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"2242","DOI":"10.1182\/blood.V76.11.2242.2242","article-title":"The molecular genetic analysis of hemophilia A: A directed search strategy for the detection of point mutations in the human factor VIII gene","volume":"76","author":"Pattinson","year":"1990","journal-title":"Blood"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"1541","DOI":"10.1111\/j.1538-7836.2011.04408.x","article-title":"Deep intronic variations may cause mild hemophilia A","volume":"9","author":"Castaman","year":"2011","journal-title":"J. Thromb. Haemost."},{"key":"ref_30","first-page":"145","article-title":"Factor 8 gene mutation spectrum of 270 patients with hemophilia A: Identification of 36 novel mutations","volume":"37","author":"Atik","year":"2020","journal-title":"Turk. J. Hematol."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"78","DOI":"10.1160\/TH09-11-0795","article-title":"Spectrum of causative mutations in patients with haemophilia A in Austria","volume":"104","author":"Reitter","year":"2010","journal-title":"Thromb. Haemost."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"275","DOI":"10.1007\/s10038-007-0238-y","article-title":"AICE-Genetics Study Group. Identification of 217 unreported mutations in the F8 gene in a group of 1,410 unselected Italian patients with hemophilia A","volume":"53","author":"Santacroce","year":"2008","journal-title":"J. Hum. Genet."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"357","DOI":"10.1055\/s-0037-1613212","article-title":"11 hemophilia A patients without mutations in the factor VIII encoding gene","volume":"88","author":"Klopp","year":"2002","journal-title":"Thromb. Haemost."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"20","DOI":"10.1182\/blood.V75.1.20.20","article-title":"A new von Willebrand factor (vWF) defect in a patient with factor VIII (FVIII) deficiency but with normal levels and multimeric patterns of both plasma and platelet vWF. Characterization of abnormal vWF\/FVIII interaction","volume":"75","author":"Mazurier","year":"1990","journal-title":"Blood"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"1937","DOI":"10.1182\/blood.V77.9.1937.1937","article-title":"The \u201cNormandy\u201d variant of von Willebrand disease: Characterization of a point mutation in the von Willebrand factor gene","volume":"77","author":"Gaucher","year":"1991","journal-title":"Blood"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"61","DOI":"10.1016\/S0092-8674(00)81146-0","article-title":"Mutations in the ER\u2013Golgi intermediate compartment protein ERGIC-53 cause combined deficiency of coagulation factors V and VIII","volume":"93","author":"Nichols","year":"1998","journal-title":"Cell"},{"key":"ref_37","first-page":"2261","article-title":"ERGIC-53 gene structure and mutation analysis in 19 combined factors V and VIII deficiency families","volume":"93","author":"Nichols","year":"1999","journal-title":"Blood"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"220","DOI":"10.1038\/ng1153","article-title":"Bleeding due to disruption of a cargo-specific ER-To-Golgi transport complex","volume":"34","author":"Zhang","year":"2003","journal-title":"Nat. Genet."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"1402","DOI":"10.1055\/s-0038-1649954","article-title":"Haemophilia A: Mutation type determines risk of inhibitor formation","volume":"74","author":"Schwaab","year":"1995","journal-title":"Thromb. Haemost."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"844","DOI":"10.1055\/s-0037-1613931","article-title":"Relationship between Factor VIII mutation type and inhibitor development in a cohort of previously untreated patients treated with recombinant factor VIII (RecombinateTM). Recombinate PUP Study Group","volume":"83","author":"Goodeve","year":"2000","journal-title":"Thromb. Haemost."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"558","DOI":"10.1046\/j.1365-2516.1998.440558.x","article-title":"Factor VIII inhibitors in mild and moderate-severity haemophilia A","volume":"4","author":"Hay","year":"1998","journal-title":"Haemophilia"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"20","DOI":"10.1016\/j.thromres.2018.05.027","article-title":"Risk factors for inhibitor development in severe hemophilia A","volume":"168","author":"Garagiola","year":"2018","journal-title":"Thromb. Res."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"405","DOI":"10.1038\/gim.2015.30","article-title":"Standards and Guidelines for the Interpretation of Sequence Variants: A Joint Consensus Recommendation of the American College of Medical Genetics and Genomics and the Association for Molecular Pathology","volume":"17","author":"Richards","year":"2015","journal-title":"Genet Med."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/25\/15\/8366\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T15:27:18Z","timestamp":1760110038000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/25\/15\/8366"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,7,31]]},"references-count":43,"journal-issue":{"issue":"15","published-online":{"date-parts":[[2024,8]]}},"alternative-id":["ijms25158366"],"URL":"https:\/\/doi.org\/10.3390\/ijms25158366","relation":{},"ISSN":["1422-0067"],"issn-type":[{"type":"electronic","value":"1422-0067"}],"subject":[],"published":{"date-parts":[[2024,7,31]]}}}