Effect of F8 B domain gene variants on synthesis, secretion, activity and stability of factor VIII protein

2014 ◽  
Vol 111 (01) ◽  
pp. 58-66 ◽  
Author(s):  
Saskia Pahl ◽  
Anna Pavlova ◽  
Julia Driesen ◽  
Johannes Oldenburg
Keyword(s):  
Intracellular Transport ◽  
Coagulation Factor ◽  
Haemophilia A ◽  
Missense Mutations ◽  
Severe Haemophilia ◽  
Wild Type ◽  
Mild Phenotype ◽  
Cell Lysate ◽  
Fviii Activity

SummaryThe B domain of the coagulation factor (F)VIII comprises some unique characteristics. Though the B domain is important for processing, intracellular transport and secretion of FVIII protein, its role in the coagulation still remains unclear. This study aims to investigate the influence of 19 reported B domain variants on quantity and quality of expressed FVIII protein. F8 variants were transiently expressed in HEK293T cells. Media and cell lysates were collected after 72 hours. FVIII synthesis, relative secretion, activity and thermostability were analysed in comparison to FVIII wild-type. Eleven of 19 analysed B domain variants showed normal FVIII activity (FVIII:C), and antigen values (40–150 %). Eight variants exhibited a decreased FVIII:C, corresponding to a mild phenotype most likely due to impaired expression and secretion mechanism, reduced thermostability or combined mechanisms. One variant, p.His1066Tyr, showed markedly reduced FVIII antigen in cell lysate. The variants p.Asp845Glu, p.His998Gln, and p.Ala1610Ser revealed a significantly decreased relative secretion. Additionally, six B domain variants significantly reduced stability of FVIII. In conclusion, none of the analysed missense mutations was causative for a severe haemophilia A (HA) phenotype. Nevertheless, the mutations p.Asp845Glu, p.Pro947Arg, p.Glu1057Lys, p.His1066Tyr, p.Arg1126Trp, p.Arg1329His, p.Leu1481Pro, and p.Ala1610Ser resulted in decreased FVIII:C values that may explain mild HA phenotypes.

scholarly journals Efficacy and safety of the drug Octofactor in prophylactic treatment in patients with severe haemophilia A

2018 ◽  
Vol 5 (3) ◽  
pp. 60-73 ◽  
Author(s):  
T. A. Andreeva ◽  
V. Yu. Zorenko ◽  
I. L. Davydkin ◽  
V. N. Konstantinova ◽  
O. E. Zalepukhina ◽  
...  
Keyword(s):  
Hemophilia A ◽  
Bleeding Episode ◽  
Coagulation Factor ◽  
Preventive Treatment ◽  
Haemophilia A ◽  
Severe Haemophilia ◽  
Efficacy And Safety ◽  
Spontaneous Bleeding ◽  
Bleeding Episodes ◽  
Blood Coagulation Factor Viii

Relevance.The development of a new recombinant blood coagulation factor VIII preparation is a promising step towards optimizing the treatment of hemophilia A. An introduction of a new medication into clinical practice precedes a clinical trials to evaluate the efficacy and safety.Materials and methods.The efficacy and safety of the domestic recombinant B-domain deleted blood coagulation factor VIII (FVIII) (moroctocog alfa, Octofactor®, JSC “GENERIUM”) were studied in the preventive treatment of 31 patients aged 21 to 52 years with severe haemophilia A. The Octofactor was administered in doses of 40 ± 5 IU/kg 3 times per week at intervals of at least 48 hours for 21 ± 1 weeks.Results.The efficacy of therapy was evaluated in 30 patients, since 1 patient refused to participate in the trial after the first injection of the study medication. There were registered 43 episodes of bleeding among 11 patients in the course of the preventive treatment with Octofactor. The average number of bleeding episodes was 1.4 ± 2.58. There were 43 bleeding episodes, 9 (20.9 %) of them were posttraumatic, 34 (79.1 %) of them were spontaneous. The average number of the spontaneous bleeding episodes (a major criterion of the efficacy) was 1.13 ± 2.19, which showed a low incidence of exacerbations of the hemorrhagic syndrome in the course of preventive treatment with Octofactor. Among all registered bleeding episodes there were 6 (14 %) mild episodes, 37 (86 %) moderate episodes. Among all spontaneous bleedings there were 6 mild episodes (17.6 %), 28 (82.4 %) moderate episodes. All posttraumatic bleedings were moderate. The vast majority (36, or 83.7 %) of bleeding episodes were stopped with administration of the Octofactor. The average number of administrations of the Octofactor for arresting 1 bleeding episode was 1.2 ± 0.56, for 1 spontaneous bleeding episode – 1.2 ± 0.59. On average, it was required to administer 3534.9 ± 2329.02 IU of the Octofactor to stop 1 episode of bleeding. In the vast majority of patients with severe hemophilia A (83.3–86.7 %),  the remaining activity FVIII was 1 % or more after the administration of the Octofactor in 48 hours. The total amount of the Octofactor, introduced for the prevention of bleeding, was 6,107,000 IU, to stop bleeding – 152,000 IU. The safety of therapy was evaluated in 31 patients. There were recorded 25 adverse events (AE) in 17 patients. Among them the laboratory ones prevailed in 23 (92 %) cases, which is not associated with the use of the trial medication. There were noted nausea and an unpleasant aftertaste in the mouth in 1 patient during the first administration of the Octofactor, and therefore he refused to continue to participate in the trial. Causality 2 AE with the study drug was regarded as definite. Such AE are expected and described in the instructions to the preparation. All AE were not serious and mild and resolved without outcomes. There were no presented thromboembolic events and immunogenic reactions.Conclusions.The obtained data testify to the efficacy and safety of the Octofactor both for preventive measures and for stopping bleeding in adult patients with severe hemophilia A.

scholarly journals RESULTS FROM A 52-WEEK, NONINTERVENTIONAL STUDY OF BRAZILIAN INDIVIDUALS WITH SEVERE HAEMOPHILIA A RECEIVING PROPHYLAXIS: RATES OF BLEEDING, FVIII USE, AND QUALITY OF LIFE

2021 ◽  
Vol 43 ◽  
pp. S232-S233
Author(s):  
GG Yamaguti-Hayakawa ◽  
PR Villaça ◽  
C Lorenzato ◽  
MH Cerqueira ◽  
LCO Oliveira ◽  
...  
Keyword(s):  
Quality Of Life ◽  
Haemophilia A ◽  
Severe Haemophilia ◽  
Noninterventional Study

scholarly journals Dental management of patients with haemophilia in the era of recombinant treatments: increased efficacy and decreased clinical risk

2019 ◽  
Vol 12 (4) ◽  
pp. e227974
Author(s):  
Antonio Liras ◽  
Luis Romeu
Keyword(s):  
Factor Viii ◽  
Conventional Treatment ◽  
Coagulation Factors ◽  
Clotting Factor ◽  
Haemophilia A ◽  
Severe Haemophilia ◽  
Optimal Result ◽  
Dental Management ◽  
Efficient Access

Haemophilia is a hereditary X-linked recessive disorder caused by a deficiency of either clotting factor VIII (haemophilia A) or IX (haemophilia B). Conventional treatment is currently based on the use of either plasma derived or recombinant coagulation factors. This paper reports on the case of a patient with severe haemophilia who presented with mesial decay and interproximal tartar build-up, for which extraction and scaling to remove tartar deposits were indicated. Following extraction, the usual haemostasis techniques were applied, and postoperative prophylactic antihaemophilic treatment was indicated for 2 or 3 days. The patient presented with moderate bleeding for a few minutes immediately after the procedure. Administration of factor VIII before surgery as well as the patient’s favourable pharmacokinetic response allowed for an optimal result. This treatment has afforded patients with haemophilia a better quality of life, and safe and efficient access to invasive surgical procedures.

Pharmacokinetics, Coagulation Factor Consumption, and Product Efficacy in Patients Crossing Over from Full-Length FVIII to B-Domain Deleted R-FVIII and Back to Full-Length FVIII.

Blood ◽  
2008 ◽  
Vol 112 (11) ◽  
pp. 2269-2269
Author(s):  
Catherine J Rea ◽  
Alex Dunkerley ◽  
Benny Sorensen ◽  
Savita Rangarajan
Keyword(s):  
Clinical Efficacy ◽  
Half Life ◽  
Coagulation Factor ◽  
Outcome Data ◽  
Full Length ◽  
Haemophilia A ◽  
Severe Haemophilia ◽  
Efficacy And Safety ◽  
Significant Difference ◽  
Recovery Times

Abstract Introduction: Treatment with B-domain deleted recombinant factor VIII (BDD-rFVIII) has proven to be effective and safe both in clinical trials and post marketing surveillance studies. However, intermittent concerns have been raised regarding the pharmacokinetic performance, efficacy and incidence of inhibitor formation with the BDD-rFVIII product. Aims: The objective of the present study was to perform a retrospective survey of half-life measurements, clinical efficacy and safety in patients with severe Haemophilia A, when switching treatment from full-length FVIII (FL-FVIII) to BDD- rFVIII and then back to full-length FVIII. We hypothesized that the biological half-life of FVIII would be equal regardless of the product used. Furthermore, we hypothesized that the total factor consumption and bleeding frequency would be indistinguishable irrespective of product used. Finally, we report on safety as evaluated by development of inhibitors and clinical outcome following surgery. Methods: Patients treated with BDD-rFVIII (between 1998 and 2008) were identified from an in-house database. Data collected included annual half-life (T/2) and recovery times (K values), total coagulation factor consumption, and number of bleeds per year as measures of clinical efficacy. Safety data consisted of surgical outcome data and incidence of inhibitor formation. The information was extracted from electronic databases and verified by a review of patients’ clinical notes. The outcome data was non-parametric, hence, paired analysis was performed using Wilcoxon signed rank test and Friedman ANOVA. Data is given as a median value and range. P-value < 0.05 was set as level of statistical significance. Results: In total, 70 patients received BDD-rFVIII on at least one occasion. Following a specified list of criteria, evaluable data was obtainable for 15 males, all with verified severe Haemophilia A (FVIII:C <1%). The average age was 10.2 years (median 10, range 4–17). The median duration on BDD-rFVIII was 30 months (range 20–54). Using the one-way non-parametric ANOVA, no statistically significant difference was detected between the half-life and recovery times recorded during the switch from FL-FVIII (T/2 median 9.15 hours, range 6.4–22; K median 2.7, range 2.0–3.4) to BDD-rFVIII (T/2 median 9.7, range 4.7–16.8; K median 1.8, range 1.0–3.5) and back to FL-FVIII (T/2 median 9.0, range 5.0–19.5; K median 2.0, range 1.6–2.8). Furthermore, there was no significant difference in coagulation factor usage (BDD-rFVIII median 4803 iu/kg/year, range 659–11304; FL-FVIII median 5349, range 1691–10146), nor number of reported bleeds (BDDrFVIII median 6, range 0–24; FL-FVIII median 5.0, range 0–17). None of the 15 patients developed an inhibitor on BDD-rFVIII. Of the total 70 patients, 11 received BDD-rFVIII to cover surgical procedures (8 minor, 3 major interventions). There were no reports of excess bleeding. Conclusions: In this retrospective survey, BDD-rFVIII was found to be equivalent to other FVIII products in terms of half-life measurements, clinical efficacy and safety.

scholarly journals Characterization of Leu777Pro and Ile865Thr type IIA von Willebrand disease mutations

Blood ◽  
1994 ◽  
Vol 83 (6) ◽  
pp. 1551-1557 ◽  
Author(s):  
SE Lyons ◽  
KA Cooney ◽  
P Bockenstedt ◽  
D Ginsburg
Keyword(s):  
Molecular Weight ◽  
Mammalian Cells ◽  
Intracellular Transport ◽  
Von Willebrand Disease ◽  
Normal Donor ◽  
Missense Mutations ◽  
Wild Type ◽  
Full Spectrum ◽  
Group I ◽  
Von Willebrand

Abstract Type IIA von Willebrand disease (vWD) is an autosomal dominant bleeding disorder characterized by a qualitative defect in von Willebrand factor (vWF). A number of missense mutations responsible for type IIA vWD have recently been identified. This report examines the type IIA vWD mutations Leu777-->Pro and Ile865-->Thr by expression of recombinant vWF containing mutant and wild-type (WT) sequences. Recombinant vWF containing the L777P mutation (vWFL777P) showed markedly impaired secretion compared with that for wild-type vWF (vWFWT) after DNA transfection into mammalian cells. Multimer analysis of secreted vWFL777P showed predominantly low molecular weight forms. In contrast, recombinant vWF containing the I865T mutation (vWFI865T) was processed in a pattern similar to vWFWT, with secretion of the full spectrum of vWF multimers. Thus, L777P and I865T are subclassified as type IIA group I and group II mutations, respectively. Analysis of platelet vWF from a patient heterozygous for the L777P mutation shows reduced large vWF multimers in a pattern similar to plasma, consistent with the intracellular transport defect predicted for a group I mutation. An increase in the proportion of high molecular weight multimers observed in type IIA vWD patient plasma, after renal transplantation from a normal donor, suggests that the kidney endothelium may be a major source of plasma vWF.

Physical fitness, functional ability and quality of life in children with severe haemophilia: a pilot study

Haemophilia ◽  
2006 ◽  
Vol 12 (5) ◽  
pp. 494-499 ◽  
Author(s):  
J. VAN DER NET ◽  
R. C. VOS ◽  
R. H. H. ENGELBERT ◽  
M. H. VAN DEN BERG ◽  
P. J. M. HELDERS ◽  
...  
Keyword(s):  
Quality Of Life ◽  
Pilot Study ◽  
Physical Fitness ◽  
Functional Ability ◽  
Haemophilia A ◽  
Severe Haemophilia

F VIII Inhibitors in Haemophilia A Patients after Continuous Infusion of Factor VIII Concentrates.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 3100-3100
Author(s):  
Ch. von Auer ◽  
J. Oldenburg ◽  
M. von Depka ◽  
C. Escuriola-Ettinghausen ◽  
W. Kreuz ◽  
...  
Keyword(s):  
Factor Viii ◽  
Continuous Infusion ◽  
Orthopaedic Surgery ◽  
Coagulation Factor ◽  
Haemophilia A ◽  
Missense Mutations ◽  
Factor Viii Concentrates ◽  
Coagulation Factor Concentrates ◽  
Major Orthopaedic Surgery ◽  
Mild Haemophilia

Abstract Continuous infusion (CI) of coagulation factor concentrates has been used since the early 1990s. Recent reports of the occurrence of an inhibitor (inh) after CI have raised concerns about this method of treatment. We conduct a retrospective study to investigate the development of inh after CI of FVIII concentrates in Germany. 99 haemophilia treating physicians in Germany were contacted and asked to answer a questionnaire. So far data of 13 departments have been reported and analyzed. Three of these 13 centers never conducted a CI, in 5 no inh were detected and in 5 haemophilia centers 10 patients with inh development after CI were registered. 5 of these patients were suffering from severe, 1 from moderate and 4 from mild haemophilia (age between 7 months and 57 years). Indications for treatment were major bleeds and surgical procedures. Plasma derived (6) and recombinant (4) factor concentrates were given in various infusion sets. Data concerning infused amount (4300 to >100000 IE), exposure days (1 to >100) and inh characteristic (alloantibodies, 3 LR, 7 HR) were collected. Regarding the genotype, we found 4 missense-mutations, 2 intron-22-inversions, 1 small deletion, 3 were unknown. In our own center we found no inh in 81 patients with major orthopaedic surgery and bolus infusion of factor VIII concentrate compared to 2 inh in 8 patients with major orthopaedic surgery and CI of FVIII. In conclusion we found only in 2 patients the typical gene mutation for inh development. Strikingly the inh developed very often in patients with mild haemophilia. These findings agree with published results. There might be an uncommon inh-pathomechanism due to CI or patients with mild haemophilia might exhibit a much higher prevalence of inhibitor development when treated with an “intensive FVIII-treatment” such as CI.

Physical function and quality of life in adolescents with haemophilia (SO-FIT study)

2013 ◽  
Vol 1 (2) ◽  
pp. 11-14 ◽  
Author(s):  
Kate Khair ◽  
Melanie Bladen ◽  
Michael Holland
Keyword(s):  
Quality Of Life ◽  
Clinical Practice ◽  
Functional Outcome ◽  
Physical Function ◽  
Mobile Devices ◽  
Haemophilia A ◽  
Severe Haemophilia ◽  
Quality Of Life Measures

Abstract Introduction: Self-completed measures of physical function and quality of life are increasingly being used in clinical practice yet little is known about how these measures correlate with joint scores in boys with severe haemophilia. In addition, it is not known whether currently used measures of functional outcome correlate with quality of life measures, which measure of physical function is most accurate and whether these measures are acceptable to a well treated contemporary cohort of boys. Methods: The Study Of physical Function In adolescenTs with haemophilia (SO-FIT) is a multicenter, randomized cross-over study designed to answer these questions, and to determine whether these self-reported measures are completed more fully and frequently if made available on mobile devices rather than with conventional pen and paper questionnaires. The study is being run by haemophilia nurse specialists and physiotherapists in UK haemophilia centres and will recruit 120 boys with severe haemophilia A or B, with or without inhibitors aged 8-16 years. Results:The SO-FIT study is now underway, and is expected to complete by the end of 2014.

Sign in / Sign up

Export Citation Format

Share Document