{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,28]],"date-time":"2026-04-28T22:56:55Z","timestamp":1777417015115,"version":"3.51.4"},"reference-count":40,"publisher":"MDPI AG","issue":"3","license":[{"start":{"date-parts":[[2024,2,1]],"date-time":"2024-02-01T00:00:00Z","timestamp":1706745600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Sanofi Company","award":["TW20002"],"award-info":[{"award-number":["TW20002"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Sporadic hemophilia A (HA) enables the persistence of HA in the population. F8 gene inversion originates mainly in male germ cells during meiosis. To date, no studies have shown the origin and timing of HA sporadic noninversion variants (NIVs); herein, we assume that HA-sporadic NIVs are generated as a de novo variant. Of the 125 registered families with HA, 22 were eligible for inclusion. We conducted a linkage analysis using F8 gene markers and amplification refractory mutation system\u2013quantitative polymerase chain reaction to confirm the origin of the sporadic NIVs (~0% mutant cells) or the presence of a mosaic variant, which requires further confirmation of the origin in the parent. Nine mothers, four maternal grandmothers, and six maternal grandfathers were confirmed to be the origin of sporadic NIVs, which most likely occurred in the zygote within the first few cell divisions and in single sperm cells, respectively. Three mothers had mosaic variants, which most likely occurred early in postzygotic embryogenesis. All maternal grandparents were free from sporadic NIV. In conclusion, F8 NIVs in sporadic HA were found to be caused primarily by de novo variants. Our studies are essential for understanding the genetic pathogenesis of HA and improving current genetic counseling.<\/jats:p>","DOI":"10.3390\/ijms25031763","type":"journal-article","created":{"date-parts":[[2024,2,1]],"date-time":"2024-02-01T08:44:08Z","timestamp":1706777048000},"page":"1763","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["De Novo Noninversion Variants Implicated in Sporadic Hemophilia A: A Variant Origin and Timing Study"],"prefix":"10.3390","volume":"25","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-5076-2917","authenticated-orcid":false,"given":"Ming","family":"Chen","sequence":"first","affiliation":[{"name":"Department of Genomic Medicine, Changhua Christian Hospital, Changhua 500, Taiwan"},{"name":"Department of Obstetrics and Gynecology, Changhua Christian Hospital, Changhua 500, Taiwan"},{"name":"Department of Medical Genetics National Taiwan University Hospital, Taipei 100, Taiwan"},{"name":"Department of Obstetrics and Gynecology, National Taiwan University Hospital, Taipei 100, Taiwan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7789-2148","authenticated-orcid":false,"given":"Ming-Ching","family":"Shen","sequence":"additional","affiliation":[{"name":"Department of Laboratory Medicine, National Taiwan University Hospital, Taipei 100, Taiwan"},{"name":"Department of Internal Medicine, National Taiwan University Hospital, Taipei 100, Taiwan"},{"name":"Hemophilia Treatment and Thrombosis Center, Department of Internal Medicine, Changhua Christian Hospital, Changhua 500, Taiwan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Shun-Ping","family":"Chang","sequence":"additional","affiliation":[{"name":"Department of Genomic Medicine, Changhua Christian Hospital, Changhua 500, Taiwan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4925-7101","authenticated-orcid":false,"given":"Gwo-Chin","family":"Ma","sequence":"additional","affiliation":[{"name":"Department of Genomic Medicine, Changhua Christian Hospital, Changhua 500, Taiwan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Dong-Jay","family":"Lee","sequence":"additional","affiliation":[{"name":"Department of Genomic Medicine, Changhua Christian Hospital, Changhua 500, Taiwan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Adeline","family":"Yan","sequence":"additional","affiliation":[{"name":"Department of Genomic Medicine, Changhua Christian Hospital, Changhua 500, Taiwan"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2024,2,1]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1111\/hae.14046","article-title":"WFH guidelines for the management of hemophilia, 3rd ed","volume":"26","author":"Srivastava","year":"2020","journal-title":"Haemophilia"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"3433","DOI":"10.1016\/S0021-9258(19)69627-4","article-title":"The role of phospholipid and factor VIIIa in the activation of bovine factor X","volume":"256","author":"Tans","year":"1981","journal-title":"J. Biol. Chem."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"17012","DOI":"10.1016\/S0021-9258(18)41886-8","article-title":"The active site of factor IXa is located far above the membrane surface and its conformation is altered upon association with factor VIIIa. A fluorescence study","volume":"267","author":"Mutucumarana","year":"1992","journal-title":"J. Biol. Chem."},{"key":"ref_4","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_5","doi-asserted-by":"crossref","first-page":"236","DOI":"10.1038\/ng1193-236","article-title":"Inversions disrupting the factor VIII gene are a common cause of severe haemophilia A","volume":"5","author":"Lakich","year":"1993","journal-title":"Nat. Genet."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"1773","DOI":"10.1093\/hmg\/2.11.1773","article-title":"Characteristic mRNA abnormality found in half the patients with severe haemophilia A is due to large DNA inversions","volume":"2","author":"Naylor","year":"1993","journal-title":"Hum. Mol. Genet."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"4851","DOI":"10.1093\/nar\/22.22.4851","article-title":"Haemophilia A: Database of nucleotide substitutions, deletions, insertions and rearrangements of the factor VIII gene. 2nd ed","volume":"22","author":"Tuddenham","year":"1994","journal-title":"Nucleic Acids Res."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"90","DOI":"10.1111\/j.1365-2516.2006.01397.x","article-title":"Prevalence of sporadic and familial haemophilia","volume":"13","author":"Kasper","year":"2007","journal-title":"Haemophilia"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"403","DOI":"10.1016\/j.jtha.2022.12.029","article-title":"Hemophilia treatment innovation: 50 years of progress and more to come","volume":"21","author":"Mannucci","year":"2023","journal-title":"J. Thromb. Haemost."},{"key":"ref_10","first-page":"1229","article-title":"Direct and indirect estimation of the sex ratio of mutation frequencies in hemophilia A","volume":"53","author":"Oldenburg","year":"1993","journal-title":"Am. J. Hum. Genet."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1035","DOI":"10.1093\/hmg\/3.7.1035","article-title":"Factor VIII gene inversions causing severe hemophilia A originate almost exclusively in male germ cells","volume":"3","author":"Rossiter","year":"1994","journal-title":"Hum. Mol. Genet."},{"key":"ref_12","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_13","doi-asserted-by":"crossref","first-page":"1064","DOI":"10.3390\/genes5041064","article-title":"Somatic mosaicism in the human genome","volume":"5","author":"Freed","year":"2014","journal-title":"Genes"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"5403","DOI":"10.1111\/jcmm.17563","article-title":"Origin and timing of de novo variants implicated in type 2 von Willebrand disease","volume":"26","author":"Chen","year":"2022","journal-title":"J. Cell Mol. Med."},{"key":"ref_15","first-page":"657","article-title":"Characterization of the factor VIII defect in 147 patients with sporadic hemophilia A: Family studies indicate a mutation type-dependent sex ratio of mutation frequencies","volume":"58","author":"Becker","year":"1996","journal-title":"Am. J. Hum. Genet."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"870","DOI":"10.1046\/j.1365-2141.1999.01631.x","article-title":"Origin of mutation in sporadic cases of haemophilia A","volume":"106","author":"Ljung","year":"1999","journal-title":"Br. J. Haematol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"291","DOI":"10.1111\/hae.13402","article-title":"Spectrum and origin of mutations in sporadic cases of haemophilia A in China","volume":"24","author":"Lu","year":"2018","journal-title":"Haemophilia"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"375","DOI":"10.1111\/hae.13975","article-title":"Genetic mosaicism in haemophilia: A practical review to help evaluate the risk of transmitting the disease","volume":"26","author":"Lannoy","year":"2020","journal-title":"Haemophilia"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"468","DOI":"10.1016\/j.tjog.2011.10.012","article-title":"Preimplantation and prenatal genetic diagnosis of aromatic L-amino acid decarboxylase deficiency with an amplification refractory mutation system-quantitative polymerase chain reaction","volume":"50","author":"Kuo","year":"2011","journal-title":"Taiwan J. Obstet. Gynecol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"299","DOI":"10.1016\/j.gene.2014.07.039","article-title":"Validating a rapid, real-time, PCR-based direct mutation detection assay for preimplantation genetic diagnosis","volume":"548","author":"Chen","year":"2014","journal-title":"Gene"},{"key":"ref_21","doi-asserted-by":"crossref","unstructured":"Chen, H.L., Lin, P.H., Chiang, Y.T., Huang, W.J., Lin, C.F., Ma, G.C., Chang, S.P., Fan, J.Y., Lin, S.Y., and Wu, C.C. (2021). Preimplantation genetic diagnosis in hereditary hearing impairment. Diagnostics, 11.","DOI":"10.3390\/diagnostics11122395"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"548","DOI":"10.1038\/nm.3519","article-title":"An ultrasensitive method for quantitating circulating tumor DNA with broad patient coverage","volume":"20","author":"Newman","year":"2014","journal-title":"Nat. Med."},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Burgstaller, J.P., Schinogl, P., Dinnyes, A., M\u00fcller, M., and Steinborn, R. (2007). Mitochondrial DNA heteroplasmy in ovine fetuses and sheep cloned by somatic cell nuclear transfer. BMC Dev. Biol., 7.","DOI":"10.1186\/1471-213X-7-141"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"9","DOI":"10.1097\/PDM.0b013e31821e59dc","article-title":"Comparison of a PNA clamp PCR and an ARMS\/Scorpion PCR assay for the detection of K-ras mutations","volume":"21","author":"Oltedal","year":"2012","journal-title":"Diagn. Mol. Pathol."},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Stadler, J., Eder, J., Pratscher, B., Brandt, S., Schneller, D., M\u00fcllegger, R., Vogl, C., Trautinger, F., Brem, G., and Burgstaller, J.P. (2015). SNPase-ARMS qPCR: Ultrasensitive Mutation-Based Detection of Cell-Free Tumor DNA in Melanoma Patients. PLoS ONE, 10.","DOI":"10.1371\/journal.pone.0142273"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"63","DOI":"10.1111\/cge.12709","article-title":"Origin of mutation in sporadic cases of severe haemophilia A in Sweden","volume":"90","author":"Ivarsson","year":"2016","journal-title":"Clin. Genet."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"75","DOI":"10.1086\/321285","article-title":"Somatic mosaicism in hemophilia A: A fairly common event","volume":"69","author":"Leuer","year":"2001","journal-title":"Am. J. Hum. Genet."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"11640","DOI":"10.1038\/s41598-021-91142-1","article-title":"Confirming putative variants at \u22645% allele frequency using allele enrichment and Sanger sequencing","volume":"11","author":"Yan","year":"2021","journal-title":"Sci. Rep."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"425","DOI":"10.2353\/jmoldx.2010.090188","article-title":"Comparison of Sanger sequencing, pyrosequencing, and melting curve analysis for the detection of KRAS mutations: Diagnostic and clinical implications","volume":"12","author":"Tsiatis","year":"2010","journal-title":"J. Mol. Diagn."},{"key":"ref_30","doi-asserted-by":"crossref","unstructured":"Wang, Y., Chen, Y., Wang, S.M., Liu, X., Gu, Y.N., and Feng, Z. (2020). Prenatal diagnosis of Duchenne muscular dystrophy revealed a novel mosaic mutation in dystrophin gene: A case report. BMC Med. Genet., 21.","DOI":"10.1186\/s12881-020-01157-0"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"231","DOI":"10.1159\/000512800","article-title":"Somatic mosaicism in the pathogenesis of de novo cerebral arteriovenous malformations: A paradigm shift implicating the RAS-MAPK signaling cascade","volume":"50","author":"Peterson","year":"2021","journal-title":"Cerebrovasc. Dis."},{"key":"ref_32","first-page":"629","article-title":"Germline origins in the human F9 gene: Frequent G:C\u2192A:T mosaicism and increased mutations with advanced maternal age","volume":"105","author":"Ketterling","year":"1999","journal-title":"Hum. Genet."},{"key":"ref_33","doi-asserted-by":"crossref","unstructured":"Cochard, L.R. (2012). Early Embryonic Development and the Placenta. Netter\u2019s Atlas of Human Embryology, Elsevier Health Sciences. [updated ed.].","DOI":"10.1016\/B978-1-4557-3977-6.00002-8"},{"key":"ref_34","unstructured":"Carlson, B.M. (2019). Human Embryology and Developmental Biology, Elsevier Health Sciences."},{"key":"ref_35","doi-asserted-by":"crossref","unstructured":"Shen, M.C., Chang, S.P., Lee, D.J., Lin, W.H., Chen, M., and Ma, G.C. (2022). Skewed X-chromosome inactivation and parental gonadal mosaicism are implicated in X-linked recessive female hemophilia patients. Diagnostics, 12.","DOI":"10.3390\/diagnostics12102267"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"787","DOI":"10.1111\/j.1365-2516.2008.01687.x","article-title":"The spectrum of the factor 8 (F8) defects in Taiwanese patients with haemophilia A","volume":"14","author":"Ma","year":"2008","journal-title":"Haemophilia"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"297","DOI":"10.1111\/j.1365-2516.2008.01866.x","article-title":"High-resolution combined linkage physical map of short tandem repeat loci on human chromosome band Xq28 for indirect haemophilia A carrier detection","volume":"15","author":"Machado","year":"2009","journal-title":"Haemophilia"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"783","DOI":"10.1111\/j.1538-7836.2010.03768.x","article-title":"Preimplantation genetic diagnosis for hemophilia A using indirect linkage analysis and direct genotyping approaches","volume":"8","author":"Laurie","year":"2010","journal-title":"J. Thromb. Haemost."},{"key":"ref_39","doi-asserted-by":"crossref","unstructured":"Acuna-Hidalgo, R., Veltman, J.A., and Hoischen, A. (2016). New insights into the generation and role of de novo mutations in health and disease. Genome Biol., 17.","DOI":"10.1186\/s13059-016-1110-1"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"29","DOI":"10.1007\/978-3-030-21664-1_2","article-title":"Age-dependent de novo mutations during spermatogenesis and their consequences","volume":"1166","author":"Cioppi","year":"2019","journal-title":"Adv. Exp. Med. Biol."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/25\/3\/1763\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T13:52:59Z","timestamp":1760104379000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/25\/3\/1763"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,2,1]]},"references-count":40,"journal-issue":{"issue":"3","published-online":{"date-parts":[[2024,2]]}},"alternative-id":["ijms25031763"],"URL":"https:\/\/doi.org\/10.3390\/ijms25031763","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,2,1]]}}}