Affinage

F8

Coagulation factor VIII · UniProt P00451

Round 2 corrected
Length
2351 aa
Mass
267.0 kDa
Annotated
2026-04-28
130 papers in source corpus 25 papers cited in narrative 24 extracted findings

Mechanistic narrative

Synthesis pass · prose summary of the discoveries below

Factor VIII (F8) is a plasma glycoprotein procofactor essential for the intrinsic pathway of blood coagulation, serving as the critical cofactor that dramatically increases the catalytic efficiency (kcat) of factor IXa for factor X activation within the intrinsic tenase complex (PMID:9834200, PMID:14684146). FVIII circulates as a metal ion-dependent (Ca²⁺, Cu²⁺) heterodimer of heavy and light chains in a noncovalent complex with von Willebrand factor (vWF), which binds via the sulfated Tyr1680 residue and the C2 domain of the FVIII light chain and protects FVIII from proteolytic degradation and immune uptake (PMID:3134349, PMID:1898735, PMID:9218428, PMID:27587878); thrombin or factor Xa activate FVIII by cleaving at Arg372, Arg740, and Arg1689, releasing it from vWF, while activated protein C inactivates FVIIIa by cleavage at Arg336 and Arg562 with consequent A2 domain dissociation (PMID:14684146, PMID:1939075). ER-to-Golgi secretion of FVIII requires the LMAN1–MCFD2 cargo receptor complex, in which MCFD2 mediates direct cargo binding while LMAN1 serves as the shuttling carrier (PMID:12717434, PMID:36490287). Loss-of-function mutations in F8—including intron 22 inversions accounting for ~45% of severe cases—cause hemophilia A, while immune tolerance to therapeutic FVIII is maintained by PD-L1⁺ regulatory T cells that eliminate FVIII-specific B cells via PD-1 ligation (PMID:8275087, PMID:36107620).

Mechanistic history

Synthesis pass · year-by-year structured walk · 13 steps
  1. 1984 High

    Cloning the full-length F8 cDNA resolved the primary structure, domain architecture, and homology to factor V and ceruloplasmin, establishing the molecular identity of the coagulation cofactor and enabling recombinant expression that corrected hemophiliac plasma clotting.

    Evidence cDNA cloning, sequencing, and expression in mammalian cells with functional clotting assay

    PMID:6438526 PMID:6438528

    Open questions at the time
    • B domain function remained unclear
    • post-translational processing pathway not yet defined
  2. 1985 High

    Identification of point mutations and deletions in the F8 gene as causes of hemophilia A established the heterogeneous mutational basis of the disease and linked genotype to phenotype.

    Evidence Southern blotting and direct sequencing in 92 hemophilia A patients

    PMID:2987704

    Open questions at the time
    • most common severe mutation (intron 22 inversion) not yet discovered
    • genotype–phenotype correlations incomplete
  3. 1991 High

    Defining the vWF-binding determinants on the FVIII light chain—particularly the essential role of sulfated Tyr1680 in the acidic region—and the APC inactivation sites (Arg336, Arg562) established the principal regulatory switches controlling FVIII stability and shutdown.

    Evidence Site-directed mutagenesis with COS-1 expression, vWF binding assays, and in vitro APC proteolysis with N-terminal sequencing of fragments

    PMID:1898735 PMID:1939075 PMID:3134349

    Open questions at the time
    • structural basis of A2 domain dissociation not determined
    • relative contribution of C2 domain to vWF binding not yet quantified
  4. 1993 High

    Discovery that intrachromosomal inversions at intron 22 account for ~45% of severe hemophilia A identified the single most common causative mutation and transformed molecular diagnostics.

    Evidence Southern blot detection of inversion-specific restriction patterns in hemophilia A cohorts

    PMID:8275087

    Open questions at the time
    • mechanism favoring inversion in male meiosis not fully elucidated
    • intron 1 inversions not yet characterized
  5. 1997 High

    Demonstrating that both the acidic region and C2 domain contribute to high-affinity vWF binding refined the two-site model for the FVIII–vWF interaction.

    Evidence SPR binding kinetics of defined FVIII light chain fragments with vWF, monoclonal antibody competition

    PMID:9218428

    Open questions at the time
    • atomic-resolution structure of the FVIII–vWF interface not determined
  6. 1998 High

    Kinetic characterization established that FVIIIa increases factor IXa activity primarily by elevating kcat rather than Km for factor X, defining the cofactor mechanism within the tenase complex.

    Evidence In vitro kinetic analysis of reconstituted tenase complex

    PMID:9834200

    Open questions at the time
    • structural basis of kcat enhancement by FVIIIa not resolved
    • role of membrane surface in cofactor function incompletely defined
  7. 2003 High

    Identification of MCFD2 mutations in combined FV/FVIII deficiency patients revealed that the LMAN1–MCFD2 complex functions as the ER-to-Golgi cargo receptor for FVIII (and FV), solving the secretion pathway question.

    Evidence Patient mutation analysis, co-immunoprecipitation, yeast two-hybrid, subcellular co-localization

    PMID:12717434

    Open questions at the time
    • relative contributions of LMAN1 vs MCFD2 to cargo binding not dissected
    • whether the complex is rate-limiting for secretion unknown
  8. 2008 High

    The X-ray crystal structure of B domain-deleted FVIII revealed the spatial arrangement of the five domains with bound Ca²⁺ and Cu²⁺ ions, and in silico docking provided the first model of the FVIIIa–FIXa interface.

    Evidence X-ray crystallography of BDD-FVIII, computational docking with FIXa

    PMID:18400180

    Open questions at the time
    • no experimental structure of the FVIIIa–FIXa complex
    • conformational changes upon thrombin activation not captured
  9. 2016 High

    Identification of an immunodominant HLA-DRB1*01:01-restricted T-cell epitope in the FVIII C2 domain (residues 2194–2213) with a remarkably narrow TCR repertoire explained a key driver of inhibitor development, and subsequent mutagenesis showed that F2196A and M2199A abolish T-cell recognition while preserving procoagulant activity.

    Evidence MHC class II tetramer staining, T-cell cloning, TCR sequencing, peptide–MHCII binding assays, recombinant FVIII mutagenesis with activity measurement

    PMID:27471234 PMID:29444872

    Open questions at the time
    • whether deimmunized FVIII variants are tolerated long-term in vivo not tested
    • epitopes restricted by other HLA alleles not fully mapped
  10. 2019 Medium

    Discovery that miRNAs (miR-374b-5p, miR-30c-5p, miR-19b-3p, miR-186-5p) target the F8 3′UTR and suppress FVIII production established a post-transcriptional regulatory layer for F8 gene expression.

    Evidence miRNA sequencing, gain- and loss-of-function experiments in FVIII-expressing cell lines, RNA-affinity purification

    PMID:31785023 PMID:32850803

    Open questions at the time
    • physiological relevance of miRNA-mediated regulation of endogenous FVIII levels not demonstrated in vivo
    • no reporter assay confirming direct 3′UTR interaction for all four miRNAs
  11. 2021 High

    Demonstration that PD-L1⁺ Tregs eliminate FVIII-specific B cells via PD-1 ligation revealed the cellular mechanism maintaining peripheral tolerance to FVIII and explained why immune tolerance induction (ITI) can resolve inhibitors.

    Evidence Mouse knockout and ITI models, human patient samples, PD-1/PD-L1 blockade, B cell apoptosis assays, flow cytometry

    PMID:36107620

    Open questions at the time
    • whether PD-L1+ Treg mechanism applies across all HLA backgrounds not established
    • signals driving PD-L1 upregulation on Tregs during ITI not identified
  12. 2021 High

    Identification of an endothelial-specific enhancer element within F8 intron 1 that drives markedly elevated FVIII levels when duplicated provided the first cis-regulatory explanation for familial thrombophilia due to FVIII excess.

    Evidence Genetic analysis of familial thrombophilia, quantitative RT-PCR, luciferase reporter assay in endothelial cells

    PMID:33275657

    Open questions at the time
    • transcription factors binding the enhancer element not identified
    • chromatin context of enhancer activity in non-endothelial cells not explored
  13. 2023 High

    Mechanistic dissection of the LMAN1–MCFD2 complex showed that MCFD2 performs direct cargo binding while LMAN1 serves as the cycling carrier, and that this complex is not rate-limiting for FVIII secretion, revising the earlier model.

    Evidence LMAN1- and MCFD2-deficient cell lines with rescue experiments and mutant LMAN1/MCFD2 overexpression across HEK293T, HepG2, and HCT116 cells

    PMID:36490287

    Open questions at the time
    • identity of the rate-limiting step in FVIII ER-to-Golgi transport remains unknown
    • structural basis of MCFD2–FVIII interaction not determined

Open questions

Synthesis pass · forward-looking unresolved questions
  • Key unresolved questions include the atomic-resolution structure of the FVIIIa–FIXa tenase complex, the identity of transcription factors binding the intron 1 enhancer, the in vivo physiological relevance of miRNA regulation of FVIII levels, and whether deimmunized FVIII variants can achieve long-term tolerance in patients.
  • no experimental FVIIIa–FIXa co-crystal or cryo-EM structure
  • cis-regulatory architecture of the F8 locus incompletely defined
  • clinical translation of deimmunized FVIII not tested

Mechanism profile

Synthesis pass · controlled-vocabulary classification · explore literature graph →
Molecular activity
GO:0098772 molecular function regulator activity 3 GO:0008289 lipid binding 1
Localization
GO:0005576 extracellular region 4 GO:0005783 endoplasmic reticulum 4 GO:0005794 Golgi apparatus 3 GO:0031410 cytoplasmic vesicle 1
Pathway
R-HSA-109582 Hemostasis 5 R-HSA-9609507 Protein localization 4 R-HSA-1643685 Disease 3 R-HSA-168256 Immune System 3
Partners
F10F9LMAN1MCFD2PROCVWF
Complex memberships
FVIII–vWF complexintrinsic tenase complex (FVIIIa–FIXa–FX)

Evidence

Reading pass · 24 per-paper findings extracted from the source corpus
Year Finding Method Journal Conf PMIDs
1984 The complete mRNA sequence encoding human coagulation factor VIII was cloned and expressed in mammalian cells, revealing a single-chain precursor of 2,351 amino acids (Mr ~267 kDa) with a domain structure showing sequence repeats and structural relatedness to factor V and ceruloplasmin; recombinant protein corrected clotting time in hemophiliac plasma. cDNA cloning, sequencing, and expression in cultured mammalian cells; functional clotting assay Nature High 6438526 6438528
1985 Point mutations (nonsense) and partial deletions in the F8 gene were identified as molecular causes of hemophilia A, establishing the heterogeneous mutational basis of the disease. Southern blotting and direct sequencing of F8 gene in 92 hemophilia A patients Nature High 2987704
1988 The FVIII light chain (80 kDa), but not the heavy chain, binds to von Willebrand factor (vWF) with a stoichiometry of one light chain per vWF subunit; thrombin cleavage removing an acidic 41-residue N-terminal peptide from the light chain completely abolishes vWF binding, and intact FVIII bound to vWF is fully released after thrombin proteolysis. Analytical velocity sedimentation of purified porcine FVIII chains with multimeric vWF; thrombin proteolysis experiments The Journal of biological chemistry High 3134349
1991 Sulfation of Tyr1680 in the acidic region of the FVIII light chain is essential for interaction with vWF; site-directed mutagenesis replacing Tyr1680 with Phe completely abolished vWF binding, and expression in the presence of chlorate (sulfation inhibitor) also abrogated binding. Site-directed mutagenesis, expression in COS-1 cells, vWF binding assay, cell-free sulfation studies with tyrosylprotein sulfotransferase The Journal of biological chemistry High 1898735
1991 Activated protein C (APC) inactivates factor VIII and factor VIIIa by proteolytic cleavage within the heavy chain at Arg336 and Arg562 (and proposed Arg740); cleavage at Arg562 most closely correlates with loss of cofactor activity and promotes dissociation of the A2 domain from the A1/light chain dimer. In vitro proteolysis of purified human FVIII/FVIIIa with APC, NH2-terminal sequencing of cleavage fragments, anti-A2 monoclonal antibody reactivity, gel filtration analysis The Journal of biological chemistry High 1939075
1993 Inversions disrupting the factor VIII gene, resulting from intrachromosomal recombination between a homologous sequence in intron 22 (int22h) and upstream copies of that sequence, account for approximately 45% of severe hemophilia A cases. Southern blot assay detecting inversion-specific restriction fragment patterns in hemophilia A patient cohorts Nature genetics High 8275087
1997 Both the acidic region (residues 1649–1689, including sulfated Tyr1680) and the C2 domain of the FVIII light chain are directly involved in forming the high-affinity vWF-binding site; the acidic region is also required to maintain the optimal conformation of the vWF-binding site within C2. Limited V8 protease digestion of FVIII light chain to generate defined fragments; surface plasmon resonance binding measurements; anti-C2 and anti-acidic region monoclonal antibody assays The Journal of biological chemistry High 9218428
1998 FVIII circulates as a metal ion-dependent heterodimer of heavy chain and light chain; activation by thrombin or factor Xa involves limited proteolysis at three sites yielding factor VIIIa, which dramatically increases the catalytic efficiency of factor IXa in activating factor X primarily by increasing kcat. Biochemical characterization review synthesizing in vitro proteolysis, activity assays, and structural data Blood High 9834200
2003 Mutations in MCFD2, an EF-hand domain protein that forms a calcium-dependent heteromeric complex with LMAN1 in the ERGIC, cause combined FV and FVIII deficiency (F5F8D), establishing that the LMAN1-MCFD2 complex functions as a cargo receptor for ER-to-Golgi transport of both FV and FVIII. Patient mutation analysis, subcellular co-localization, co-immunoprecipitation demonstrating calcium-dependent MCFD2-LMAN1 interaction, yeast two-hybrid Nature genetics High 12717434
2004 Factor VIIIa acts as a cofactor by markedly increasing the catalytic rate constant (kcat) of factor IXa for factor X activation; thrombin or factor Xa activate FVIII by cleaving at three defined sites in the heavy chain (Arg372, Arg740) and light chain (Arg1689), altering covalent structure and conformation. In vitro proteolysis with thrombin/FXa, kinetic assays of FIXa/FVIIIa complex activity, fragment characterization Blood reviews High 14684146
2008 X-ray crystal structure of B domain-deleted human factor VIII revealed five globular domains with one Ca2+ and two Cu2+ ions; A1 and A3 domains form the base of a triangular A-domain heterotrimer; C1 and C2 domains contain membrane-binding features; in silico docking with factor IXa suggested an extended interface spanning both heavy and light chains of FVIII. X-ray crystallography (crystal structure determination) of BDD-FVIII; in silico docking modeling with factor IXa based on biochemical constraints Structure High 18400180
2009 The LMAN1-MCFD2 complex serves as a cargo receptor for ER-to-Golgi transport of FV and FVIII; MCFD2 missense mutations in EF-hand domains abolish the calcium-dependent interaction with LMAN1; the B domain of FVIII may be important in mediating its interaction with the LMAN1-MCFD2 complex. Patient mutation analysis in F5F8D, biochemical characterization of MCFD2-LMAN1 interaction, review of functional studies British journal of haematology High 19183188
2011 LMAN1-deficient mice show ~50% reduction in plasma FV and FVIII and platelet FV, confirming the ER-to-Golgi cargo receptor role of the LMAN1-MCFD2 complex for FV and FVIII; ER in Lman1-/- hepatocytes is distended with accumulation of α1-antitrypsin and GRP78, indicating ER stress. Mouse knockout model (Lman1-/- mice), plasma factor activity assays, electron microscopy, immunohistochemistry, in vitro COPII vesicle formation assay Blood High 21795745
2013 Human mesenchymal stem cells (MSC) from lung, liver, brain, and bone marrow express FVIII mRNA and secrete functional FVIII protein; in MSC, FVIII localizes to the perinuclear region rather than being stored in granules, in contrast to endothelial cells. Quantitative RT-PCR, confocal microscopy with FVIII-specific antibody, aPTT and chromogenic functional assays of MSC supernatants and lysates Journal of cellular physiology Medium 23042590
2006 Regulated secretion of both VWF and FVIII from endothelial storage granules (Weibel-Palade bodies) occurs only when there is endogenous co-synthesis of FVIII together with VWF; VWF serves as the carrier protecting FVIII from proteolytic degradation in plasma. DDAVP stimulation experiments, cell biology studies of VWF/FVIII co-expression and co-storage Pediatric blood & cancer Medium 16470522
2016 VWF inhibits uptake of FVIII by immature dendritic cells and activation of FVIII-specific T cells in a dose-dependent manner; recombinant VWF lacking the FVIII-binding domain did not inhibit T-cell activation, indicating that VWF reduces FVIII immunogenicity by shielding it from antigen-presenting cells through direct binding. In vitro dendritic cell uptake assay, T-cell activation assay with VWF and VWF mutant lacking FVIII-binding domain Blood Medium 27587878
2019 miR-374b-5p and miR-30c-5p target the 3'UTR of F8 mRNA; overexpression of these miRNAs in cell lines constitutively expressing FVIII decreased FVIII expression, while an miR-30c-5p inhibitor partially restored FVIII expression, establishing a miRNA-based mechanism for F8 gene regulation. miRNA sequencing, overexpression of miRNAs in FVIII-expressing cell lines, miRNA inhibitor experiments, FVIII activity assay Transfusion Medium 31785023
2020 miR-19b-3p and miR-186-5p directly interact with F8 mRNA (identified by RNA-affinity purification) and suppress FVIII protein levels when overexpressed in mammalian cells, providing further evidence that miRNAs targeting the F8 3'UTR can modulate FVIII production. In vivo RNA-affinity purification, miRNA overexpression in mammalian cells, FVIII activity measurement Frontiers in cell and developmental biology Medium 32850803
2023 LMAN1 primarily serves as a shuttling carrier for MCFD2, while MCFD2 carries out the actual cargo binding and transport of FV and FVIII; LMAN1 carbohydrate-binding activity is not essential for FV/FVIII transport; overexpression of MCFD2 alone is sufficient to rescue FV/FVIII secretion in LMAN1-deficient cells; the LMAN1-MCFD2 complex is not rate-limiting for ER-Golgi transport of FV/FVIII. LMAN1- and MCFD2-deficient cell lines (HEK293T, HepG2, HCT116), FV/FVIII secretion assays, rescue experiments with wild-type and mutant LMAN1/MCFD2 overexpression Blood advances High 36490287
2021 Immune tolerance against FVIII is maintained by PD-L1-expressing regulatory T cells (Tregs) that ligate PD-1 on FVIII-specific B cells causing their apoptosis; FVIII-deficient mice lack such Tregs and develop inhibitors; repetitive FVIII injection (ITI) induces FVIII-specific PD-L1+ Tregs and re-engages B cell elimination; FVIII-specific Tregs exist in humans and upregulate PD-L1 after successful ITI. Mouse knockout model, flow cytometry, ITI mouse model, human patient samples, PD-1/PD-L1 blockade experiments, B cell apoptosis assays The Journal of clinical investigation High 36107620
2021 A 23.4-kb tandem duplication of the proximal F8 gene (promoter, exon 1, and part of intron 1) causes markedly elevated FVIII levels (>400%) and familial thrombophilia; the duplication produces twofold upregulation of F8 mRNA; a 927-bp region within the duplicated F8 intron 1 contains an enhancer element driving >45-fold increased reporter activity in endothelial cells. Genetic analysis, quantitative RT-PCR, luciferase reporter assay in endothelial cells, chromatin accessibility analysis Blood High 33275657
2016 A single immunodominant HLA-DRA*01-DRB1*01:01-restricted epitope in FVIII (peptide 2194–2213, C2 domain) is recognized by CD4+ T-effector cells from both severe and mild hemophilia A subjects with inhibitors; high-avidity T-cell clones from multiple subjects share the same T-cell receptor beta (TCRB) gene, indicating a remarkably narrow TCR repertoire driving inhibitor responses. MHC class II tetramer staining, T-cell clone isolation, cytokine secretion assays, TCRB gene sequencing, high-throughput immunosequencing Blood High 27471234
2018 Amino acid substitution F2196A or M2199A within the immunodominant FVIII C2-domain epitope (residues 2194–2205) abrogates HLA-DRB1*01:01-restricted T-cell proliferation while retaining normal procoagulant activity and expression levels, establishing these residues as critical for MHC class II binding and T-cell recognition. Peptide-MHCII binding assays, T-cell proliferation assays with clones and polyclonal lines, production of recombinant BDD-FVIII mutant proteins with FVIII activity measurement Blood advances High 29444872
2010 A point mutation causing Leu176Pro substitution in the A1 domain of rat FVIII (F8 gene, autosomal chromosome 18 in rats) disrupts the tertiary structure of FVIII and causes hemophilia A-like FVIII deficiency; the defect is corrected by human plasma or recombinant human FVIII administration. F8 cDNA sequencing, coagulation factor activity assays, structural prediction, human FVIII replacement experiment in WAG/RijYcb rats Journal of thrombosis and haemostasis Medium 20626616

Source papers

Stage 0 corpus · 130 papers · ranked by NIH iCite citations
Year Title Journal Citations PMID
2002 Generation and initial analysis of more than 15,000 full-length human and mouse cDNA sequences. Proceedings of the National Academy of Sciences of the United States of America 1479 12477932
1984 Molecular cloning of a cDNA encoding human antihaemophilic factor. Nature 834 6438528
2012 Genetically distinct subsets within ANCA-associated vasculitis. The New England journal of medicine 726 22808956
1993 Inversions disrupting the factor VIII gene are a common cause of severe haemophilia A. Nature genetics 689 8275087
2004 The human plasma proteome: a nonredundant list developed by combination of four separate sources. Molecular & cellular proteomics : MCP 658 14718574
2011 Phylogenetic-based propagation of functional annotations within the Gene Ontology consortium. Briefings in bioinformatics 656 21873635
1984 Expression of active human factor VIII from recombinant DNA clones. Nature 620 6438526
2003 The dynamics of thrombin formation. Arteriosclerosis, thrombosis, and vascular biology 387 12524220
2005 Human plasma N-glycoproteome analysis by immunoaffinity subtraction, hydrazide chemistry, and mass spectrometry. Journal of proteome research 350 16335952
1998 The life cycle of coagulation factor VIII in view of its structure and function. Blood 333 9834200
2010 Novel associations of multiple genetic loci with plasma levels of factor VII, factor VIII, and von Willebrand factor: The CHARGE (Cohorts for Heart and Aging Research in Genome Epidemiology) Consortium. Circulation 295 20231535
2012 F8 gene mutation type and inhibitor development in patients with severe hemophilia A: systematic review and meta-analysis. Blood 287 22282501
2003 Bleeding due to disruption of a cargo-specific ER-to-Golgi transport complex. Nature genetics 234 12717434
1991 Sulfation of Tyr1680 of human blood coagulation factor VIII is essential for the interaction of factor VIII with von Willebrand factor. The Journal of biological chemistry 220 1898735
2005 Polymorphisms in the IL10 but not in the IL1beta and IL4 genes are associated with inhibitor development in patients with hemophilia A. Blood 212 16380445
2011 Toward an understanding of the protein interaction network of the human liver. Molecular systems biology 207 21988832
1985 Detection and sequence of mutations in the factor VIII gene of haemophiliacs. Nature 203 2987704
1988 Association of the factor VIII light chain with von Willebrand factor. The Journal of biological chemistry 202 3134349
2008 Crystal structure of human factor VIII: implications for the formation of the factor IXa-factor VIIIa complex. Structure (London, England : 1993) 185 18400180
2013 Factor VIII gene (F8) mutation and risk of inhibitor development in nonsevere hemophilia A. Blood 181 23926300
2016 Life in the shadow of a dominant partner: the FVIII-VWF association and its clinical implications for hemophilia A. Blood 180 27587878
2003 No evidence of association between prothrombotic gene polymorphisms and the development of acute myocardial infarction at a young age. Circulation 177 12615788
1998 Factor VIII inhibitors in mild and moderate-severity haemophilia A. UK Haemophilia Centre Directors Organisation. Thrombosis and haemostasis 174 9569189
1991 Activated protein C-catalyzed inactivation of human factor VIII and factor VIIIa. Identification of cleavage sites and correlation of proteolysis with cofactor activity. The Journal of biological chemistry 169 1939075
2004 Activation of factor VIII and mechanisms of cofactor action. Blood reviews 165 14684146
2009 Gene-centric association signals for lipids and apolipoproteins identified via the HumanCVD BeadChip. American journal of human genetics 164 19913121
1997 The acidic region of the factor VIII light chain and the C2 domain together form the high affinity binding site for von willebrand factor. The Journal of biological chemistry 159 9218428
2012 Distinct characteristics of antibody responses against factor VIII in healthy individuals and in different cohorts of hemophilia A patients. Blood 158 23243272
2010 Codon optimization of human factor VIII cDNAs leads to high-level expression. Blood 157 21041718
1987 FVIII inhibitor IgG subclass and FVIII polypeptide specificity determined by immunoblotting. Blood 155 2436689
1991 Molecular characterization of severe hemophilia A suggests that about half the mutations are not within the coding regions and splice junctions of the factor VIII gene. Proceedings of the National Academy of Sciences of the United States of America 153 1908096
2011 F8 and F9 mutations in US haemophilia patients: correlation with history of inhibitor and race/ethnicity. Haemophilia : the official journal of the World Federation of Hemophilia 110 22103590
2003 Efficient production of human FVIII in hemophilic mice using lentiviral vectors. Molecular therapy : the journal of the American Society of Gene Therapy 95 12718905
1999 Conditional stability of the HemA protein (glutamyl-tRNA reductase) regulates heme biosynthesis in Salmonella typhimurium. Journal of bacteriology 87 9973348
1989 Cloning, genetic characterization, and nucleotide sequence of the hemA-prfA operon of Salmonella typhimurium. Journal of bacteriology 85 2544564
2005 Resin monomer 2-hydroxyethyl methacrylate (HEMA) is a potent inducer of apoptotic cell death in human and mouse cells. Journal of dental research 83 15668336
2014 HEMA-induced cytotoxicity: oxidative stress, genotoxicity and apoptosis. International endodontic journal 69 24355064
2005 Pattern of cell death after in vitro exposure to GDMA, TEGDMA, HEMA and two compomer extracts. Dental materials : official publication of the Academy of Dental Materials 69 16223522
1989 Isolation, nucleotide sequence, and preliminary characterization of the Escherichia coli K-12 hemA gene. Journal of bacteriology 69 2548996
1990 Cloning and characterization of the hemA region of the Bacillus subtilis chromosome. Journal of bacteriology 59 2110138
2019 Curing hemophilia A by NHEJ-mediated ectopic F8 insertion in the mouse. Genome biology 55 31843008
2008 Gene transfer to hemophilia A mice via oral delivery of FVIII-chitosan nanoparticles. Journal of controlled release : official journal of the Controlled Release Society 55 18634839
2015 Parameters influencing FVIII pharmacokinetics in patients with severe and moderate haemophilia A. Haemophilia : the official journal of the World Federation of Hemophilia 54 25582282
2004 Antioxidative vitamins decrease cytotoxicity of HEMA and TEGDMA in cultured cell lines. Archives of oral biology 53 14693206
2009 Recent developments in the understanding of the combined deficiency of FV and FVIII. British journal of haematology 48 19183188
2014 Diagnosis and management challenges in patients with mild haemophilia A and discrepant FVIII measurements. Haemophilia : the official journal of the World Federation of Hemophilia 45 24517184
1993 5-Aminolevulinic acid availability and control of spectral complex formation in hemA and hemT mutants of Rhodobacter sphaeroides. Journal of bacteriology 45 8468291
2011 Mice deficient in LMAN1 exhibit FV and FVIII deficiencies and liver accumulation of α1-antitrypsin. Blood 44 21795745
1989 Isolation and nucleotide sequence of the hemA gene of Escherichia coli K12. Molecular & general genetics : MGG 44 2664455
2009 Immune response to FVIII in hemophilia A: an overview of risk factors. Clinical reviews in allergy & immunology 42 19148784
2005 Inhibitor development in patients with hemophilia A after continuous infusion of FVIII concentrates. Annals of the New York Academy of Sciences 42 16126990
2017 Noninvasive detection of F8 int22h-related inversions and sequence variants in maternal plasma of hemophilia carriers. Blood 41 28490568
2008 Combined FV and FVIII deficiency. Haemophilia : the official journal of the World Federation of Hemophilia 41 19141160
2007 HEMA reduces cell proliferation and induces apoptosis in vitro. Dental materials : official publication of the Academy of Dental Materials 40 17920114
1993 G6PD haplotypes spanning Xq28 from F8C to red/green color vision. Genomics 40 8104869
2019 Synthesis and Characterization of Silicone Contact Lenses Based on TRIS-DMA-NVP-HEMA Hydrogels. Polymers 38 31159172
2006 Regulated release of VWF and FVIII and the biologic implications. Pediatric blood & cancer 38 16470522
1996 5-Aminolevulinate production by Escherichia coli containing the Rhodobacter sphaeroides hemA gene. Applied and environmental microbiology 38 8837411
2020 Electrospun GelMA fibers and p(HEMA) matrix composite for corneal tissue engineering. Materials science & engineering. C, Materials for biological applications 34 33545871
2019 Cell Loaded GelMA:HEMA IPN hydrogels for corneal stroma engineering. Journal of materials science. Materials in medicine 32 31811387
2010 Expression, engineering and characterization of the tumor-targeting heterodimeric immunocytokine F8-IL12. Protein engineering, design & selection : PEDS 32 20551083
2002 The identification and classification of 41 novel mutations in the factor VIII gene (F8C). Human mutation 32 11857744
1987 Recombination between the factor VIII gene and the DXS52 locus gives the most probable genetic order as centromere-fra(X)-DXS15-DXS52-F8C-telomere. American journal of medical genetics 32 3105317
1995 Regulation of the hemA gene during 5-aminolevulinic acid formation in Pseudomonas aeruginosa. Journal of bacteriology 31 7883699
2015 Assessment of HEMA and TEGDMA induced DNA damage by multiple genotoxicological endpoints in human lymphocytes. Dental materials : official publication of the Academy of Dental Materials 30 26025483
2016 Targeting of the human F8 at the multicopy rDNA locus in Hemophilia A patient-derived iPSCs using TALENickases. Biochemical and biophysical research communications 28 26921444
2016 T cells from hemophilia A subjects recognize the same HLA-restricted FVIII epitope with a narrow TCR repertoire. Blood 28 27471234
2021 Partial F8 gene duplication (factor VIII Padua) associated with high factor VIII levels and familial thrombophilia. Blood 26 33275657
2008 Expression of a hemA gene from Agrobacterium radiobacter in a rare codon optimizing Escherichia coli for improving 5-aminolevulinate production. Applied biochemistry and biotechnology 26 18800199
2002 The Pseudomonas aeruginosa hemA promoter is regulated by Anr, Dnr, NarL and Integration Host Factor. Molecular genetics and genomics : MGG 26 12073043
2023 Thermoresponsive Smart Copolymer Coatings Based on P(NIPAM-co-HEMA) and P(OEGMA-co-HEMA) Brushes for Regenerative Medicine. ACS biomaterials science & engineering 25 37874897
2013 Mesenchymal stem cells contribute to endogenous FVIII:c production. Journal of cellular physiology 25 23042590
2019 Clinical manifestation of hemophilia A in the absence of mutations in the F8 gene that encodes FVIII: role of microRNAs. Transfusion 24 31785023
2018 FVIII proteins with a modified immunodominant T-cell epitope exhibit reduced immunogenicity and normal FVIII activity. Blood advances 24 29444872
2015 Phenotype-genotype correlations in hemophilia A carriers are consistent with the binary role of the phase between F8 and X-chromosome inactivation. Journal of thrombosis and haemostasis : JTH 24 25611311
2013 Stem cell suspension injected HEMA-lactate-dextran cryogels for regeneration of critical sized bone defects. Artificial cells, nanomedicine, and biotechnology 24 23477355
2013 Characterization of large deletions in the F8 gene using multiple competitive amplification and the genome walking technique. Journal of thrombosis and haemostasis : JTH 24 23551875
2020 B Cell Depletion Eliminates FVIII Memory B Cells and Enhances AAV8-coF8 Immune Tolerance Induction When Combined With Rapamycin. Frontiers in immunology 23 32670285
2014 Fc-fusion technology and recombinant FVIII and FIX in the management of the hemophilias. Drug design, development and therapy 23 24729686
2020 The Immune Response to the fVIII Gene Therapy in Preclinical Models. Frontiers in immunology 22 32351497
2023 Comprehensive Analysis of Hemophilia A (CAHEA): Towards Full Characterization of the F8 Gene Variants by Long-Read Sequencing. Thrombosis and haemostasis 21 37285902
2019 Solvent and HEMA Increase Adhesive Toxicity and Cytokine Release from Dental Pulp Cells. Materials (Basel, Switzerland) 21 31461952
2015 Cell toxicity of 2-hydroxyethyl methacrylate (HEMA): the role of oxidative stress. European journal of oral sciences 21 25968591
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