Spectrum of F8 Genotype and Genetic Impact on Inhibitor Development in Patients with Hemophilia A from Multicenter Cohort Studies (J-HIS) in Japan

Abstract

Some genetic and treatment-related factors are risk factors for inhibitor development in patients with hemophilia A (PwHA). However, the genotype distribution of the factor VIII gene (F8) and genetic impact on inhibitor development in Japanese PwHA remain unknown. In 2007, the Japan Hemophilia Inhibitor Study 2 (J-HIS2) was organized to establish a nationwide registry system for hemophiliacs and to elucidate risk factors for inhibitor development, designed for prospective investigation following a retrospective study (J-HIS1) which had already finished. Patients, newly diagnosed after January 2007, were enrolled in J-HIS2 and followed up for inhibitor development and clinical environments since 2008 onward. In the present study, F8 genotypes of PwHA were investigated in the patients recruited from the J-HIS2 cohort as well as those with inhibitor from the J-HIS1 cohort. F8 variants identified in 59 PwHA with inhibitor in J-HIS1 were: 20 intron-22 inversions, 5 intron-1 inversions, 9 large deletions, 4 nonsense, 8 missense, 11 small in/del, and 2 splice-site variants. F8 variants identified in 267 (67 with inhibitor) PwHA in J-HIS2 were: 76(28) intron–22 inversions, 3(2) intron–1 inversion, 1(0) duplication, 8(5) large deletions, 21(7) nonsense, 109(7) missense, 40(11) small in/del, and 9(7) splice-site variants. Forty variants were novel. The cumulative inhibitor incidence rate in the severe group with null changes was 42.4% (95% confidence interval [CI]: 33.7–50.8), higher than that with nonnull changes (15.6% [95%CI: 6.8–27.8]), in J-HIS2. Relative risk for inhibitor development of null changes was 2.89. The spectrum of F8 genotype and genetic impact on inhibitor development in Japanese PwHA were consistent with the previous reports.

Keywords

hemophilia A - inhibitor - F8 - null variants - nonnull variants

Study Group of J-HIS1 and J-HIS2 Consisting of Steering Committee and Collaborative Institutes

Steering Committee Members of J-HIS 1 and J-HIS 2

Teruhisa Fujii (Hiroshima University, Hiroshima), Katsuyuki Fukutake* (Tokyo Medical University, Tokyo), Isao Hamaguchi (National Institute of Infectious Diseases, Tokyo), Hideji Hanabusa** (Ogikubo Hospital, Tokyo), Satoshi Higasa (Hyogo College of Medicine, Hyogo), Yasuo Horikoshi (Shizuoka Children's Hospital, Shizuoka), Masue Imaizumi (Miyagi Children's Hospital, Miyagi), Fuminori Iwasaki (Kanagawa Children's Medical Center, Kanagawa), Kiyoshi Kawakami (Kagoshima City Hospital, Kagoshima), Masao Kobayashi (Hiroshima University, Hiroshima), Tetsuhito Kojima (Nagoya University, Aichi), Takeshi Matsumoto (Mie University Hospital, Mie), Yoko Matsuo (Kurume University Hospital, Fukuoka), Tadashi Matsushita (Nagoya University, Aichi), Hisaya Nakadate (National Center for Child Health and Development, Tokyo), Keiji Nogami (Nara Medical University, Nara), Shouichi Ohga (Kyushu University, Fukuoka), Yoshitoshi Otsuka (Hyogo College of Medicine, Hyogo), Toshiaki Oka (Sapporo Tokushukai Hospital, Hokkaido), Yuri Okimoto (Chiba Children's Hospital, Chiba), Michio Sakai (Hospital of the University of Occupational and Environmental Health, Fukuoka), Midori Shima (Nara Medical University, Nara), Keiko Shinozawa (Tokyo Medical University, Tokyo), Akira Shirahata (Kitakyushu Yahata Higashi Hospital, Fukuoka), Takashi Suzuki (Ogikubo Hospital, Tokyo), Junki Takamatsu (Japan Red Cross Aichi blood Center, Aichi), Hideyuki Takedani (The Institute of Medical Science, The University of Tokyo, Tokyo), Masashi Taki (St. Marianna University School of Medicine Yokohama City Seibu Hospital, Kanagawa), Maiko Taneichi (National Institute of Infectious Diseases, Tokyo), Koji Yada (Nara Medical University, Nara), and Akira Yoshioka (Nara Medical University, Nara).

*K.F. was a program supervisor of AMED and has resigned the committee before initiation of a grant from AMED in 2017. **We would like to offer our deepest sympathies to Dr. Hideji Hanabusa who passed away untimely in October 2016.

Collaborative Institutes (65)

Aichi Sannomaru Hospital (Aichi), Anjo Kosei Hospital (Aichi), Nagoya University (Aichi), Akita University Hospital (Akita), Odate Municipal General Hospital (Akita), Aihara Internal Medical Clinic (Aomori), Kuroishi General Hospital (Aomori), Kameda General Hospital (Chiba), Matsudo City General Hospital (Chiba), Ehime University Hospital (Ehime), Tsuruga Medical Center (Fukui), Fukuoka University Hospital (Fukuoka), Hospital of the University of Occupational and Environmental Health Japan (Fukuoka), Kurume University Hospital (Fukuoka), Kyushu University Hospital (Fukuoka), Hiroshima University Hospital (Hiroshima), Asahikawa Medical University Hospital (Hokkaido), Iwamizawa Municipal General Hospital (Hokkaido), Kushiro Red Cross Hospital (Hokkaido), Sapporo Hokuyu Hospital (Hokkaido), Sapporo Medical University Hospital (Hokkaido), Sapporo Tokushukai Hospital (Hokkaido), Hyogo College of Medicine (Hyogo), Hyogo Prefectural Amagasaki General Medical Center (Hyogo), Kobe University Hospital (Hyogo), Ibaraki Children's Hospital (Ibaraki), Iwate Prefectural Ofunato Hospital (Iwate), Shikoku Medical Center for Children and Adults (Kagawa), Kagoshima City Hospital (Kagoshima), St. Marianna University School of Medicine Yokohama City Seibu Hospital (Kanagawa), St. Marianna University School of Medicine Hospital (Kanagawa), Japanese Red Cross Kochi Hospital (Kochi), Japanese Red Cross Kumamoto Hospital (Kumamoto), Ayabe City Hospital (Kyoto), Kyoto University Hospital (Kyoto), Mie University Hospital (Mie), Hasegawa Pediatric Clinic (Miyagi), Miyagi Children's Hospital (Miyagi), Sendai Medical Center (Miyagi), Sendai Nishitaga Hospital (Miyagi), Tohoku University Hospital (Miyagi), University of Miyazaki Hospital (Miyazaki), Japanese Red Cross Nagano Hospital (Nagano), Nara Medical University (Nara), Niigata Cancer Center Hospital (Niigata), Oita Memorial Hospital (Oita), Oita Prefectural Hospital (Oita), Kitano Hospital (Osaka), Osaka Rosai Hospital (Osaka), Saga University Hospital (Saga), Japanese Red Cross Otsu Hospital (Shiga), Seirei Hamamatsu General University (Shizuoka), Shizuoka Children's Hospital (Shizuoka), Dokkyo Medical University Hospital (Tochigi), Japanese Red Cross Ashikaga Hospital (Tochigi), National Center for Child Health and Development (Tokyo), National Center for Global Health and Medicine (Tokyo), Ogikubo Hospital (Tokyo), Teikyo University Hospital (Tokyo), University of Tokyo, Institute of Medical Science (Tokyo), Tokyo Medical University (Tokyo), Tokyo Metropolitan Children's Medical Center (Tokyo), Japanese Red Cross Wakayama Medical Center, (Wakayama), Naga Municipal Hospital (Wakayama), Tsuruoka Municipal Shonai Hospital (Yamagata).

Note

This study was presented in an abstract form at the Congress of World Federation of Hemophilia, Orlando, Florida, United States, July 2016, and the Congress of International Society on Thrombosis and Haemostasis (ISTH), Melbourne, Australia, July 2019.

Authors' Contributions

K.S.: genotyping, clinical laboratory testing, analyzed data, interpreted the data, wrote manuscript; K.Y.: clinical work, blood sampling, genotyping, interpreted the data, made figures and tables, analyzed data, wrote manuscript; T.K.: clinical work, blood sampling, advised the study, and edited manuscript; K.N.: clinical work, interpreted the data, wrote and edited manuscript; M.T.: clinical work, blood sampling; K.F.: clinical work, blood sampling, advised the study, study design, interpretation of data, and revision of the manuscript; A.Y. and A.S.: organized and initiated the study; M.S.: clinical work, supervised the study. K.S. and K.Y. equally contributed to this work, and both of them have the right to list herself (himself) first in bibliography.

^* These authors contributed equally to this study as first authors.

Publication History

Received: 05 June 2020

Accepted: 21 October 2020

Article published online:
30 November 2020

• References

• 1 White II GC, Rosendaal F, Aledort LM, Lusher JM, Rothschild C, Ingerslev J. Factor VIII and Factor IX Subcommittee. Definitions in hemophilia. Recommendation of the scientific subcommittee on factor VIII and factor IX of the scientific and standardization committee of the International Society on Thrombosis and Haemostasis. Thromb Haemost 2001; 85 (03) 560
• 2 Gouw SC, van den Berg HM, Fischer K. et al; PedNet and Research of Determinants of INhibitor development (RODIN) Study Group. Intensity of factor VIII treatment and inhibitor development in children with severe hemophilia A: the RODIN study. Blood 2013; 121 (20) 4046-4055
• 3 Gouw SC, Van Der Bom JG, Van Den Berg HM, Zewald RA, Ploos Van Amstel JK, Mauser-Bunschoten EP. Influence of the type of F8 gene mutation on inhibitor development in a single centre cohort of severe haemophilia A patients. Haemophilia 2011; 17 (02) 275-281
• 4 Astermark J, Wang X, Oldenburg J, Berntorp E, Lefvert AK. MIBS Study Group. Polymorphisms in the CTLA-4 gene and inhibitor development in patients with severe hemophilia A. J Thromb Haemost 2007; 5 (02) 263-265
• 5 Astermark J, Oldenburg J, Pavlova A, Berntorp E, Lefvert AK. MIBS Study Group. Polymorphisms in the IL10 but not in the IL1beta and IL4 genes are associated with inhibitor development in patients with hemophilia A. Blood 2006; 107 (08) 3167-3172
• 6 Gouw SC, van den Berg HM, Oldenburg J. et al. F8 gene mutation type and inhibitor development in patients with severe hemophilia A: systematic review and meta-analysis. Blood 2012; 119 (12) 2922-2934
• 7 Eckhardt CL, van Velzen AS, Peters M. et al; INSIGHT Study Group. Factor VIII gene (F8) mutation and risk of inhibitor development in nonsevere hemophilia A. Blood 2013; 122 (11) 1954-1962
• 8 Tuddenham EG, McVey JH. The genetic basis of inhibitor development in haemophilia A. Haemophilia 1998; 4 (04) 543-545
• 9 McVey JH, Rallapalli PM, Kemball-Cook G. et al. The European Association for Haemophilia and Allied Disorders (EAHAD) coagulation factor variant databases: Important resources for haemostasis clinicians and researchers. Haemophilia 2020; 26 (02) 306-313
• 10 Lakich D, Kazazian Jr HH, Antonarakis SE, Gitschier J. Inversions disrupting the factor VIII gene are a common cause of severe haemophilia A. Nat Genet 1993; 5 (03) 236-241
• 11 Bagnall RD, Waseem N, Green PM, Giannelli F. Recurrent inversion breaking intron 1 of the factor VIII gene is a frequent cause of severe hemophilia A. Blood 2002; 99 (01) 168-174
• 12 Project entrusted by Ministry of Health, Labour And Welfare. Nationwide Survey on Coagulation Disorders 2019. Published by Japan Foundation for AIDS Prevention. Accessed November 5, 2020 at: https://api-net.jfap.or.jp/library/alliedEnt/02/backNumber.html
• 13 Inaba H, Shibata H, Yoshida S. et al. Inversion in Japanese patients with hemophilia A. Thromb Haemost 1995; 74 (02) 810
• 14 Fukuda K, Naka H, Morichika S. et al. Inversions of the factor VIII gene in Japanese patients with severe hemophilia A. Int J Hematol 2004; 79 (03) 303-306
• 15 Yada K, Nogami K, Kawamura T, Minami H, Shima M. The first case of int1h-related inversion in Japanese haemophilia A patients. Haemophilia 2014; 20 (06) e408-e410
• 16 Iioka F, Shimomura D, Nakamura F. et al. Long-term treatment course of a patient with mild haemophilia A who developed a high titre factor VIII inhibitor. Haemophilia 2014; 20 (06) e402-e404
• 17 Yada K, Nogami K, Takeyama M, Ogiwara K, Wakabayashi H, Shima M. Mild hemophilia A patient with novel Pro1809Leu mutation develops an anti-C2 antibody inhibiting allogeneic but not autologous factor VIII activity. J Thromb Haemost 2015; 13 (10) 1843-1853
• 18 Shirahata A, Fukutake K, Higasa S. et al; Study Group on Factors Involved in Formation of Inhibitors to Factor VIII and IX Preparations. An analysis of factors affecting the incidence of inhibitor formation in patients with congenital haemophilia in Japan. Haemophilia 2011; 17 (05) 771-776
• 19 den Dunnen JT, Dalgleish R, Maglott DR. et al. HGVS Recommendations for the Description of Sequence Variants: 2016 Update. Hum Mutat 2016; 37 (06) 564-569
• 20 Kanda Y. Investigation of the freely available easy-to-use software 'EZR' for medical statistics. Bone Marrow Transplant 2013; 48 (03) 452-458
• 21 Spena S, Garagiola I, Cannavò A. et al; SIPPET Study Group. Prediction of factor VIII inhibitor development in the SIPPET cohort by mutational analysis and factor VIII antigen measurement. J Thromb Haemost 2018; 16 (04) 778-790
• 22 Lu Y, Xin Y, Dai J. et al. Spectrum and origin of mutations in sporadic cases of haemophilia A in China. Haemophilia 2018; 24 (02) 291-298
• 23 Kim HJ, Chung HS, Kim SK. et al. Mutation spectrum and inhibitor risk in 100 Korean patients with severe haemophilia A. Haemophilia 2012; 18 (06) 1008-1013
• 24 Lin SY, Su YN, Hung CC. et al. Mutation spectrum of 122 hemophilia A families from Taiwanese population by LD-PCR, DHPLC, multiplex PCR and evaluating the clinical application of HRM. BMC Med Genet 2008; 9: 53
• 25 Johnsen JM, Fletcher SN, Huston H. et al. Novel approach to genetic analysis and results in 3000 hemophilia patients enrolled in the My Life, Our Future initiative. Blood Adv 2017; 1 (13) 824-834
• 26 Payne AB, Miller CH, Kelly FM, Michael Soucie J, Craig Hooper W. The CDC Hemophilia A Mutation Project (CHAMP) mutation list: a new online resource. Hum Mutat 2013; 34 (02) E2382-E2391
• 27 Byers PH. Killing the messenger: new insights into nonsense-mediated mRNA decay. J Clin Invest 2002; 109 (01) 3-6
• 28 Nakaya S, Liu ML, Thompson AR. Some factor VIII exon 14 frameshift mutations cause moderately severe haemophilia A. Br J Haematol 2001; 115 (04) 977-982
• 29 Young M, Inaba H, Hoyer LW, Higuchi M, Kazazian Jr HH, Antonarakis SE. Partial correction of a severe molecular defect in hemophilia A, because of errors during expression of the factor VIII gene. Am J Hum Genet 1997; 60 (03) 565-573
• 30 Onsori H, Hosseinpour MA, Montaser-Kouhsari S, Asgharzadeh M, Hosseinpour AA. Identification of a novel missense mutation in exon 4 of the human factor VIII gene associated with sever hemophilia A patient. Pak J Biol Sci 2007; 10 (23) 4299-4302
• 31 Citron M, Godmilow L, Ganguly T, Ganguly A. High throughput mutation screening of the factor VIII gene (F8C) in hemophilia A: 37 novel mutations and genotype-phenotype correlation. Hum Mutat 2002; 20 (04) 267-274
• 32 Liu ML, Nakaya S, Thompson AR. Non-inversion factor VIII mutations in 80 hemophilia A families including 24 with alloimmune responses. Thromb Haemost 2002; 87 (02) 273-276
• 33 Bogdanova N, Markoff A, Pollmann H. et al. Spectrum of molecular defects and mutation detection rate in patients with severe hemophilia A. Hum Mutat 2005; 26 (03) 249-254
• 34 Guillet B, Lambert T, d'Oiron R. et al. Detection of 95 novel mutations in coagulation factor VIII gene F8 responsible for hemophilia A: results from a single institution. Hum Mutat 2006; 27 (07) 676-685
• 35 Tagariello G, Belvini D, Salviato R. et al. Experience of a single Italian center in genetic counseling for hemophilia: from linkage analysis to molecular diagnosis. Haematologica 2000; 85 (05) 525-529
• 36 Bicocchi MP, Pasino M, Lanza T. et al. Analysis of 18 novel mutations in the factor VIII gene. Br J Haematol 2003; 122 (05) 810-817
• 37 Lannoy N, Abinet I, Bosmans A, Lambert C, Vermylen C, Hermans C. Computational and molecular approaches for predicting unreported causal missense mutations in Belgian patients with haemophilia A. Haemophilia 2012; 18 (03) e331-e339
• 38 Fay PJ. Activation of factor VIII and mechanisms of cofactor action. Blood Rev 2004; 18 (01) 1-15
• 39 Soeda T, Nogami K, Nishiya K. et al. The factor VIIIa C2 domain (residues 2228-2240) interacts with the factor IXa Gla domain in the factor Xase complex. J Biol Chem 2009; 284 (06) 3379-3388
• 40 Shen BW, Spiegel PC, Chang CH. et al. The tertiary structure and domain organization of coagulation factor VIII. Blood 2008; 111 (03) 1240-1247
• 41 Pratt KP, Shen BW, Takeshima K, Davie EW, Fujikawa K, Stoddard BL. Structure of the C2 domain of human factor VIII at 1.5 A resolution. Nature 1999; 402 (6760): 439-442
• 42 Stoylova SS, Lenting PJ, Kemball-Cook G, Holzenburg A. Electron crystallography of human blood coagulation factor VIII bound to phospholipid monolayers. J Biol Chem 1999; 274 (51) 36573-36578
• 43 Nogami K, Freas J, Manithody C, Wakabayashi H, Rezaie AR, Fay PJ. Mechanisms of interactions of factor X and factor Xa with the acidic region in the factor VIII A1 domain. J Biol Chem 2004; 279 (32) 33104-33113
• 44 Takeyama M, Nogami K, Sasai K, Furukawa S, Shima M. Contribution of factor VIII A2 domain residues 400-409 to a factor X-interactive site in the factor Xase complex. Thromb Haemost 2018; 118 (05) 830-841
• 45 Takeyama M, Wakabayashi H, Fay PJ. Factor VIII light chain contains a binding site for factor X that contributes to the catalytic efficiency of factor Xase. Biochemistry 2012; 51 (03) 820-828
• 46 Shepherd AJ, Skelton S, Sansom CE, Gomez K, Moss DS, Hart DP. A large-scale computational study of inhibitor risk in non-severe haemophilia A. Br J Haematol 2015; 168 (03) 413-420
• 47 Ikeda N, Kojima H, Nishikawa M. et al. Determination of HLA-A, -C, -B, -DRB1 allele and haplotype frequency in Japanese population based on family study. Tissue Antigens 2015; 85 (04) 252-259