Shortened Lifespan and Lethal Hemorrhage in a Hemophilia A Mouse Model

Staber, Janice M.; Pollpeter, Molly

doi:10.1371/journal.pone.0154857

Cited by 7 publications

(7 citation statements)

References 13 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Both have a shortened lifespan attributed to hemorrhagic events. 26 However, like the other coagulation factor knockouts, the severity was greater in mice, with ∼40% overall survival at one year 26 versus ∼75% at the same timepoint in our study. Concordantly, while FVIII-deficient mice exhibit impaired clot formation when challenged, we were surprised to find that zebrafish larvae demonstrated no visible hemorrhage and normal hemostasis when challenged with endothelial injury.…”

Section: Discussionsupporting

confidence: 41%

“…Hemophilia A mouse models exhibit reduced survival and exhibit excessive bleeding after injury. 25,26 An F8 -/rat model created using genome editing zinc finger nucleases demonstrates spontaneous bleeding including central nervous system hemorrhage, which is the most concerning bleeding in hemophilia A patients. 27,28 Vwf-deficient mice exhibit VWD, including spontaneous hemorrhage, prolonged bleeding times, and reduced FVIII levels and activity.…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Loss of factor VIII in zebrafish rebalances antithrombin deficiency but has a limited bleeding diathesis

Richter,

Raghunath,

Griffin

et al. 2024

Preprint

View full text Add to dashboard Cite

Deficiencies in coagulation factor VIII (FVIII, F8) result in the bleeding disorder hemophilia A. An emerging novel therapeutic strategy for bleeding disorders is to enhance hemostasis by limiting natural anticoagulants, such as antithrombin (AT3). To study pro/anticoagulant hemostatic balance in an in vivo model, we used genome editing to create null alleles for f8 and von Willebrand factor (vwf) in zebrafish, a model organism with a high degree of homology to the mammalian hemostatic system and unique attributes, including external development and optical transparency. f8 homozygous mutant larvae surprisingly formed normal thrombi when subjected to laser-mediated endothelial injury, had no overt signs of hemorrhage, but had a modest increase in mortality. We have previously shown that at3-/- larvae develop disseminated intravascular coagulation (DIC), with spontaneous thrombosis and fibrinogen consumption, resulting in bleeding phenotype marked by secondary lack of induced thrombus formation upon endothelial injury. We found that with loss of FVIII (f8-/-;at3-/-), larvae no longer developed spontaneous fibrin thrombi and did produce clots in response to endothelial injury. However, homozygous loss of zebrafish Vwf failed to rescue the at3 DIC phenotype. These studies demonstrate an altered balance of natural anticoagulants that mitigates FVIII deficiency in zebrafish, similar to human clinical pipeline products. The data also suggest that zebrafish FVIII might circulate independently of Vwf. Further study of this unique balance could provide new insights for management of hemophilia A and von Willebrand disease.

show abstract

Section: Discussionsupporting

confidence: 41%

Section: Introductionmentioning

confidence: 99%

Loss of factor VIII in zebrafish rebalances antithrombin deficiency but has a limited bleeding diathesis

Richter,

Raghunath,

Griffin

et al. 2024

Preprint

View full text Add to dashboard Cite

show abstract

“…Given this enhanced in vitro clotting activity, we also determined the in vivo clotting activity of FVIII-ΔF with 2 distinct hemostatic challenges in severe HA mice: the tail-clip and ferric chloride (FeCl 3 ) assays. These animals have undetectable circulating murine FVIII activity (< 1% of normal) and have excessive, often fatal, bleeding in response to trauma, which despite gentle handling, results in shortened life spans, comparable with humans with severe hemophilia without treatment (39). HA mice received 10 μg/kg of either FVIII-ΔF or FVIII-BDD purified protein immediately prior to both challenges.…”

Section: Resultsmentioning

confidence: 99%

Circumventing furin enhances factor VIII biological activity and ameliorates bleeding phenotypes in hemophilia models

et al. 2016

View full text Add to dashboard Cite

Processing by the proprotein convertase furin is believed to be critical for the biological activity of multiple proteins involved in hemostasis, including coagulation factor VIII (FVIII). This belief prompted the retention of the furin recognition motif (amino acids 1645-1648) in the design of B-domain-deleted FVIII (FVIII-BDD) products in current clinical use and in the drug development pipeline, as well as in experimental FVIII gene therapy strategies. Here, we report that processing by furin is in fact deleterious to FVIII-BDD secretion and procoagulant activity. Inhibition of furin increases the secretion and decreases the intracellular retention of FVIII-BDD protein in mammalian cells. Our new variant (FVIII-ΔF), in which this recognition motif is removed, efficiently circumvents furin. FVIII-ΔF demonstrates increased recombinant protein yields, enhanced clotting activity, and higher circulating FVIII levels after adeno-associated viral vector-based liver gene therapy in a murine model of severe hemophilia A (HA) compared with FVIII-BDD. Moreover, we observed an amelioration of the bleeding phenotype in severe HA dogs with sustained therapeutic FVIII levels after FVIII-ΔF gene therapy at a lower vector dose than previously employed in this model. The immunogenicity of FVIII-ΔF did not differ from that of FVIII-BDD as a protein or a gene therapeutic. Thus, contrary to previous suppositions, FVIII variants that can avoid furin processing are likely to have enhanced translational potential for HA therapy.

show abstract

“…This study could not explain the development of hemorrhage only in some rats (<20%). Shortened lifespan and lethal hemorrhage have been reported in a hemophilia A mouse model; however, actual mortality due to hemorrhage is constant after birth, and the survival period reaches 2 years at longest 27 . It is considered that the occurrence of severe bleeding in animals with potential coagulation abnormalities can depend on both endogenous pathophysiological background and exogenous factors (e.g., activity of individual animals).…”

Section: Discussionmentioning

confidence: 99%

Analyses of hemorrhagic diathesis in high-iron diet-fed rats

et al. 2021

View full text Add to dashboard Cite

Iron overload has been well recognized to cause oxidant-mediated cellular/tissue injury; however, little is known about the effects of iron overload on the blood coagulation system. We encountered an unexpected bleeding tendency in rats fed a high-iron diet in a set of studies using iron-modified diets. In this study, we investigated the mechanism of hemorrhagic diathesis induced by dietary iron overload in rats. Six-week-old F344/DuCrlCrlj male rats were fed a standard (containing 0.02% iron) or a high-iron diet (containing 1% iron) for 6 weeks and were then sampled for hematological, blood biochemical, coagulation, and pathological examinations. Serum and liver iron levels increased in rats fed the high-iron diet (Fe group) and serum transferrin was almost saturated with iron. However, serum transaminase levels did not increase. Moreover, plasma prothrombin time and activated partial thromboplastin time were significantly prolonged, regardless of the presence of hemorrhage. The activity of clotting factors II and VII (vitamin K-dependent coagulation factors) decreased significantly, whereas that of factor VIII was unaltered. Blood platelet levels were not influenced by dietary iron overload, suggesting that the bleeding tendency in iron-overloaded rats is caused by secondary hemostasis impairment. In addition, hemorrhage was observed in multiple organs in rats fed diets containing more than 0.8% iron. Our results suggest that iron overload can increase the susceptibility of coagulation abnormalities caused by latent vitamin K insufficiency.

show abstract

Shortened Lifespan and Lethal Hemorrhage in a Hemophilia A Mouse Model

Cited by 7 publications

References 13 publications

Loss of factor VIII in zebrafish rebalances antithrombin deficiency but has a limited bleeding diathesis

Loss of factor VIII in zebrafish rebalances antithrombin deficiency but has a limited bleeding diathesis

Circumventing furin enhances factor VIII biological activity and ameliorates bleeding phenotypes in hemophilia models

Analyses of hemorrhagic diathesis in high-iron diet-fed rats

Contact Info

Product

Resources

About