Targeted mutation of zebrafish fga models human congenital afibrinogenemia

Fish, Richard J.; Sanza, Corinne Di; Neerman-Arbez, Marguerite

doi:10.1182/blood-2013-12-547182

Cited by 31 publications

(51 citation statements)

References 16 publications

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“…In contrast to other coagulation protein deficiencies , loss of fga results in a relatively mild hemorrhagic phenotype in zebrafish. As seen previously , we also did not detect grossly visible bleeding by o‐dianisidine staining in fga − / − larval mutants (data not shown). At 41 dpf juvenile fga − / − and fga +/+ siblings were randomly selected for histological analyses.…”

Section: Resultssupporting

confidence: 89%

Loss of fibrinogen in zebrafish results in an asymptomatic embryonic hemostatic defect and synthetic lethality with thrombocytopenia

Lavik

Liu

et al. 2019

Journal of Thrombosis and Haemostasis

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Loss of fibrinogen in zebrafish has been previously shown to result in adult onset hemorrhage Hemostatic defects were discovered in early fga−/− embryos but well tolerated until adulthood Afibrinogenemia and thrombocytopenia results in synthetic lethality in zebrafish. Testing human FGA variants of uncertain significance in zebrafish identified causative mutations Summary BackgroundMutations in the alpha chain of fibrinogen (FGA), such as deficiencies in other fibrinogen subunits, lead to rare inherited autosomal recessive hemostatic disorders. These range from asymptomatic to catastrophic life‐threatening bleeds and the molecular basis of inherited fibrinogen deficiencies is only partially understood. Zinc finger nucleases have been used to produce mutations in zebrafish fga, resulting in overt adult‐onset hemorrhage and reduced survival. ObjectivesTo determine the age of onset of hemostatic defects in afibrinogenemic zebrafish and model human fibrinogen deficiencies. MethodsTALEN genome editing (transcription activator‐like effector nucleases) was used to generate a zebrafish fga mutant. Hemostatic defects were assessed through survival, gross anatomical and histological observation and laser‐induced endothelial injury. Human FGA variants with unknown pathologies were engineered into the orthologous positions in zebrafish fga. ResultsLoss of Fga decreased survival and resulted in synthetic lethality when combined with thrombocytopenia. Zebrafish fga mutants exhibit a severe hemostatic defect by 3 days of life, but without visible hemorrhage. Induced thrombus formation through venous endothelial injury was completely absent in mutant embryos and larvae. This hemostatic defect was restored by microinjection of wild‐type fga cDNA plasmid or purified human fibrinogen. This system was used to determine whether unknown human variants were pathological by engineering them into fga. ConclusionsThese studies confirm that loss of fibrinogen in zebrafish results in the absence of hemostasis from the embryonic period through adulthood. When combined with thrombocytopenia, zebrafish exhibit synthetic lethality, demonstrating that thrombocytes are necessary for survival in response to hemorrhage.

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Section: Resultssupporting

confidence: 89%

Loss of fibrinogen in zebrafish results in an asymptomatic embryonic hemostatic defect and synthetic lethality with thrombocytopenia

Lavik

Liu

et al. 2019

Journal of Thrombosis and Haemostasis

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“…157,162 The recent introduction of the 146 Gregory et al, 147 Hughes et al, 154 Day et al, 155 Khandekar and Jagadeeswaran, 156 Tournoij et al, 158 Williams et al, 159 and O'Connor et al 161 …”

Section: Future Directionsmentioning

confidence: 99%

Animal Models of Thrombosis From Zebrafish to Nonhuman Primates

Jagadeeswaran

Cooley

Gross

et al. 2016

Circulation Research

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Thrombosis is a leading cause of morbidity and mortality worldwide. Animal models are used to understand the pathological pathways involved in thrombosis and to test the efficacy and safety of new antithrombotic drugs. In this review, we will first describe the central role a variety of animal models of thrombosis and hemostasis has played in the development of new antiplatelet and anticoagulant drugs. These include the widely used P2Y 12 antagonists and the recently developed orally available anticoagulants that directly target factor Xa or thrombin. Next, we will describe the new players, such as polyphosphate, neutrophil extracellular traps, and microparticles, which have been shown to contribute to thrombosis in mouse models, particularly venous thrombosis models. Other mouse studies have demonstrated roles for the factor XIIa and factor XIa in thrombosis. This has spurred the development of strategies to reduce their levels or activities as a new approach for preventing thrombosis. Finally, we will discuss the emergence of zebrafish as a model to study thrombosis and its potential use in the discovery of novel factors involved in thrombosis and hemostasis. Animal models of thrombosis from zebrafish to nonhuman primates are vital in identifying pathological pathways of thrombosis that can be safely targeted with a minimal effect on hemostasis. Future studies should focus on understanding the different triggers of thrombosis and the best drugs to prevent each type of thrombotic event. (Circ Res.

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“…A knockout of fga using ZFNs has been described [41•], and complete ablation of fibrinogen was demonstrated using a panel of anti-zebrafish fibrinogen antibodies. Overt hemorrhage was observed in adult homozygous mutant fish but not in embryos and larvae suggesting variable roles for fibrinogen during development.…”

Section: Conservation Of the Coagulation Cascade In Zebrafishmentioning

confidence: 99%

Modeling Disorders of Blood Coagulation in the Zebrafish

2015

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Hemostasis, the process of blood clot formation and resolution in response to vascular injury, and thrombosis, the dysregulation of hemostasis leading to pathological clot formation, are widely studied. However, the genetic variability in hemostatic and thrombotic disorders is incompletely understood, suggesting that novel mediators have yet to be uncovered. The zebrafish is developing into a powerful in vivo model to study hemostasis, and its features as a model organism are well suited to (a) develop high-throughput screens to identify novel mediators of hemostasis and thrombosis, (b) validate candidate genes identified in human populations, and (c) characterize the structure/function relationship of gene products. In this review, we discuss conservation of the zebrafish hemostatic system, highlight areas for future study, and outline the utility of this model to study blood coagulation and its dysregulation.

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Targeted mutation of zebrafish fga models human congenital afibrinogenemia

Cited by 31 publications

References 16 publications

Loss of fibrinogen in zebrafish results in an asymptomatic embryonic hemostatic defect and synthetic lethality with thrombocytopenia

Loss of fibrinogen in zebrafish results in an asymptomatic embryonic hemostatic defect and synthetic lethality with thrombocytopenia

Animal Models of Thrombosis From Zebrafish to Nonhuman Primates

Modeling Disorders of Blood Coagulation in the Zebrafish

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