Summary Bypassing therapy is essential for the haemostatic management of patients with haemophilia A with inhibitor ( PWHA ‐inh), but the therapeutic effects are inconsistent. We previously reported that activated prothrombin complex concentrates ( aPCC ) activated factor (F) VIII in vitro, and was mediated mainly by the activated FVII ( FVII a) contained in aPCC . We have extended those studies to assess global coagulation in whole blood from 18 PWHA ‐inh in the co‐presence of aPCC and FVIII using Ca 2+ ‐triggered rotational thromboelastometry. The clot times ( CT s) in the presence of both aPCC (0·05 iu/ml) and recombinant (r) FVIII (1 iu/ml) ex vivo were shortened compared to the aPCC alone ( P < 0·01). These enhancing effects of rFVIII were observed, irrespective of recognizing inhibitor epitopes; however, the clot formation time and ‘α’‐angle were not significantly different. In samples from 7 PWHA ‐inh post‐infusion of aPCC (70‐80 iu/kg), only the CT s were shortened in the presence of rFVIII ex vivo compared to its absence ( P < 0·05), indicating that the enhanced activity centred on the initiation phase of coagulation. Furthermore, experiments in the co‐presence of rFVII a and rFVIII demonstrated that FVIII accelerated only the CT s. We concluded that FVIII / FVII a‐related coagulation mechanism enhanced global haemostatic function by the co‐presence of bypassing agents and FVIII in PWHA‐inh.

Thromboelastometry

10.1111/bjh.15209

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Citations (4)

Bleeding and safety outcomes in persons with haemophilia A without inhibitors: Results from a prospective non‐interventional study in a real‐world setting

Haemophilia (2019)

Rebecca Kruse‐Jarres Johannes Oldenburg Elena Santagostino Midori Shima Christine L. Kempton

Introduction Prospectively collected real‐world data on bleeds, haemophilia treatment and safety in persons with haemophilia A (PwHA) without factor VIII (FVIII) inhibitors are limited. A global, non‐interventional study (NIS; NCT02476942) prospectively collected real‐world data in PwHA who were treated per local routine clinical practice. Aim Assess annualized bleeding rate (ABR), haemophilia treatment practices and adverse events (AEs) in adult/adolescent PwHA without inhibitors. Methods Eligible participants aged ≥12 years with severe HA without history of inhibitors prospectively collected bleeding and treatment information. Results Ninety‐four participants were enrolled (median [range] age, 34 [12‐76] years) and monitored for a median (range) of 29.8 (12.4‐47.7) weeks. In the episodic (n = 45) and prophylactic (n = 49) treatment groups, respectively, 872/1066 (81.8%) and 151/189 (79.9%), bleeds were treated; ABRs (95% confidence interval) were 36.1 (30.8‐42.3) and 5.0 (3.3‐7.5), respectively, for treated bleeds and 43.1 (36.5‐50.9) and 6.2 (4.2‐9.2), respectively, for all bleeds, and median (interquartile range) ABRs were 31.1 (19.8‐51.6) and 1.9 (0.0‐8.2), respectively, for treated bleeds and 35.3 (21.7‐62.9) and 2.7 (0.0‐9.4), respectively, for all bleeds. Half of the participants on FVIII prophylaxis had relatively high adherence to treatment, using 2.9 and 2.1 median doses/wk of standard and extended half‐life FVIII, respectively. Serious AEs included gastrointestinal polyp haemorrhage and haemarthrosis; the most common AE was viral upper respiratory tract infection. Conclusion PwHA without inhibitors continue to bleed on prophylaxis, consistent with the literature, and require treatment for breakthrough bleeds. This prospective NIS demonstrates the need for more efficacious haemostatic approaches.

Interquartile range

10.1111/hae.13655

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Citations (36)

Properties of Factor VIII in the recombinant Factor VIII Concentrates (BAY w 6240)

Japanese Journal of Thrombosis and Hemostasis (1990)

Akira Yoshioka Ichiro Tanaka Midori Shima Toshiharu FUJIWARA S Terada

Properties of factor VIII (FVIII) in recombinant FVIII concentrates (rFVIII, BAY w 6240) were investigated using 6 kinds of monoclonal antibodies to FVIII.The FVIII: C levels obtained by the one-stage method using severe hemophilia A plasma as a substrate were identical with those indicated on the label (100u/ml). The FVIII: C levels obtained by the chromogenic method were relatively higher than those by the one-stage method.The FVIII: Ag levels assayed by polyclonal ELISA agreed with those by NMC-VIII/1 (recognizes the 80KDa fragment of L-chain) or NMC-VIII/5 (the 70KDa of L-chain) monoclonal ELISA, whereas the FVIII: Ag levels by C 5 (the 54KDa of H-chain) monoclonal ELISA were 2.5-3.0 times higher than those by other monoclonal ELISAs. The ratio between FVIII: Ag and FVIII: C was 1.4. It was considered that FVIII in the recombinant concentrates lost its activity less than FVIII in the heat-treated concentrates commercially available.No von Willebrand factor was detectable by the sandwich ELISA system. SDS-PAGE pattern showed almost no contamination except albumin as a stabilizer.Immunoblot analysis revealed that FVIII in recombinant concentrates could react not only with monoclonal antibodies to L-chain and H-chain, but also with monoclonal antibodies to middle portion of FVIII molecule.These results suggest that the fundamental structure of FVIII molecule for coagulant activity is preserved in the rFVIII concentrates, and that it is expected to be useful for the treatment of hemophiliacs.

Polyclonal antibodies

Chromogenic

10.2491/jjsth.1.212

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Delivery of Full-Length Factor VIII Using a piggyBac Transposon Vector to Correct a Mouse Model of Hemophilia A

PLoS ONE (2014)

Hideto Matsui Naoko Fujimoto Noriko Sasakawa Yasuhide Ohinata Midori Shima

Viral vectors have been used for hemophilia A gene therapy. However, due to its large size, full-length Factor VIII (FVIII) cDNA has not been successfully delivered using conventional viral vectors. Moreover, viral vectors may pose safety risks, e.g., adverse immunological reactions or virus-mediated cytotoxicity. Here, we took advantages of the non-viral vector gene delivery system based on piggyBac DNA transposon to transfer the full-length FVIII cDNA, for the purpose of treating hemophilia A. We tested the efficiency of this new vector system in human 293T cells and iPS cells, and confirmed the expression of the full-length FVIII in culture media using activity-sensitive coagulation assays. Hydrodynamic injection of the piggyBac vectors into hemophilia A mice temporally treated with an immunosuppressant resulted in stable production of circulating FVIII for over 300 days without development of anti-FVIII antibodies. Furthermore, tail-clip assay revealed significant improvement of blood coagulation time in the treated mice.piggyBac transposon vectors can facilitate the long-term expression of therapeutic transgenes in vitro and in vivo. This novel gene transfer strategy should provide safe and efficient delivery of FVIII.

Factor IX

HEK 293 cells

Transduction (biophysics)

10.1371/journal.pone.0104957

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Citations (48)

FVIII Heavy Chain Enhances Tenase Activity Induced By FVIIIa Mimicking Bispesific Antibody, ACE910

Blood (2014)

Hiroaki Minami Keiji Nogami Takehisa Kitazawa Kunihiro Hattori Midori Shima

Factor IXa

Sepharose

Prothrombinase

Factor IX

10.1182/blood.v124.21.1481.1481

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Effect of Depolymerized Holothurian Glycosaminoglycan (DHG) on the Activation of Factor VIII and Factor V by Thrombin

The Journal of Biochemistry (1996)

Hideki Nagase Keiichi Enjyoji Midori Shima K. Kitazato Akira Yoshioka

Our previous study has shown that depolymerized holothurian glycosaminoglycan (DHG) has two different inhibitory activities in the blood coagulation cascade: heparin cofactor II-dependent thrombin inhibition; and antithrombin III- and heparin cofactor II-indepen-dent inhibition of the intrinsic factor Xase complex [Nagase et al. (1995) Blood 85,15271534]. In the present study, the effect of DHG on the activation of factor VIII and factor V by thrombin was examined with purified human components. DHG inhibited the activation of factor VIII by thrombin at concentrations exceeding 80 nM, but not the activation of factor V by thrombin at concentrations of up to 8 μM. On Western blot analysis, DHG inhibited the cleavage of factor VIII light chain at concentrations exceeding 0.8 μM. The interaction between DHG and factors VIII and V and thrombin was examined with a DHG-cellulofine column. DHG had strong affinity for factor V and thrombin, but slight affinity for factor VIII. The interaction of DHG with thrombin was analyzed, using fluorescein isothiocyan-ate-labeled DHG. One mole of DHG bound 2 mol of thrombin, with a dissociation constant (Kd) of 3.04 ×10−6 M. These results suggest that DHG interferes with the interaction between thrombin and factor VTII, probably by making a binary complex through the anionic binding exosite II of thrombin.

Factor (programming language)

Factor IX

10.1093/oxfordjournals.jbchem.a021217

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Citations (27)

Comparison of the properties of phospholipid surfaces formed on HPA and L1 biosensor chips for the binding of the coagulation factor VIII

Journal of Chromatography A (2001)

Evgueni L. Saenko Andrey G. Sarafanov Natalya M. Ananyeva Evan Behre Midori Shima

Prothrombinase

Factor V

10.1016/s0021-9673(01)00601-x

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Citations (35)