Thrombin-deficient mutant of medaka, a model fish, displays serious retardation in blood coagulation

ABSTRACT At the last stage of the blood coagulation cascade, thrombin plays a central role in the processing of fibrinogen for the polymerization and in the additional activation of Factor XIII for the stable cross-linking of fibrin. In addition, thrombin carries out possible multiple roles via processing or interaction with various functional proteins. Several studies conducted in order to elucidate additional physiological significance are ongoing. To clarify further significance of thrombin and to establish an associated disease model, we characterized the orthologue gene for medaka (Oryzias latipes), a research model fish. Tissue distribution of medaka prothrombin has been immunotechnically analyzed. Furthermore, thrombin-deficient medaka mutants were viably established by utilizing a genome-editing method. The established gene-deficient mutants exhibited retarded blood coagulation even in the heterozygous fish. Taking advantage of their ease of handling, this specific model is useful for further investigation in medical research areas on human coagulation diseases.

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Differential Drug Target Selection in Blood Coagulation: What can we get from Computational Systems Biology Models?

Current Pharmaceutical Design ◽

10.2174/1381612826666200406091807 ◽

2020 ◽

Vol 26 (18) ◽

pp. 2109-2115 ◽

Cited By ~ 1

Author(s):

Mikhail A. Panteleev ◽

Anna A. Andreeva ◽

Alexey I. Lobanov

Keyword(s):

Blood Coagulation ◽

Biological Network ◽

Target Selection ◽

Anticoagulation Therapy ◽

Differential Regulation ◽

Coagulation Cascade ◽

Computational Systems Biology ◽

Analysis Strategies ◽

Optimal Target ◽

Computational Systems

Discovery and selection of the potential targets are some of the important issues in pharmacology. Even when all the reactions and the proteins in a biological network are known, how does one choose the optimal target? Here, we review and discuss the application of the computational methods to address this problem using the blood coagulation cascade as an example. The problem of correct antithrombotic targeting is critical for this system because, although several anticoagulants are currently available, all of them are associated with bleeding risks. The advantages and the drawbacks of different sensitivity analysis strategies are considered, focusing on the approaches that emphasize: 1) the functional modularity and the multi-tasking nature of this biological network; and 2) the need to normalize hemostasis during the anticoagulation therapy rather than completely suppress it. To illustrate this effect, we show the possibility of the differential regulation of lag time and endogenous thrombin potential in the thrombin generation. These methods allow to identify the elements in the blood coagulation cascade that may serve as the targets for the differential regulation of this system.

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Introduction to the blood coagulation cascade and cloning of blood coagulation factors

Journal of Protein Chemistry ◽

10.1007/bf01025423 ◽

1986 ◽

Vol 5 (4) ◽

pp. 247-253 ◽

Cited By ~ 11

Author(s):

Earl W. Davie

Keyword(s):

Blood Coagulation ◽

Coagulation Factors ◽

Coagulation Cascade ◽

Blood Coagulation Factors

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A genome-wide screen uncovers multiple roles for mitochondrial nucleoside diphosphate kinase D in inflammasome activation

Science Signaling ◽

10.1126/scisignal.abe0387 ◽

2021 ◽

Vol 14 (694) ◽

pp. eabe0387

Author(s):

Orna Ernst ◽

Jing Sun ◽

Bin Lin ◽

Balaji Banoth ◽

Michael G. Dorrington ◽

...

Keyword(s):

Nucleoside Diphosphate Kinase ◽

Multiple Roles ◽

Metabolic Reprogramming ◽

Inflammasome Activation ◽

Ros Production ◽

Genome Wide ◽

A Genome ◽

Mouse Macrophages ◽

Genome Scale ◽

Mitochondrial Dna Synthesis

Noncanonical inflammasome activation by cytosolic lipopolysaccharide (LPS) is a critical component of the host response to Gram-negative bacteria. Cytosolic LPS recognition in macrophages is preceded by a Toll-like receptor (TLR) priming signal required to induce transcription of inflammasome components and facilitate the metabolic reprograming that fuels the inflammatory response. Using a genome-scale arrayed siRNA screen to find inflammasome regulators in mouse macrophages, we identified the mitochondrial enzyme nucleoside diphosphate kinase D (NDPK-D) as a regulator of both noncanonical and canonical inflammasomes. NDPK-D was required for both mitochondrial DNA synthesis and cardiolipin exposure on the mitochondrial surface in response to inflammasome priming signals mediated by TLRs, and macrophages deficient in NDPK-D had multiple defects in LPS-induced inflammasome activation. In addition, NDPK-D was required for the recruitment of TNF receptor–associated factor 6 (TRAF6) to mitochondria, which was critical for reactive oxygen species (ROS) production and the metabolic reprogramming that supported the TLR-induced gene program. NDPK-D knockout mice were protected from LPS-induced shock, consistent with decreased ROS production and attenuated glycolytic commitment during priming. Our findings suggest that, in response to microbial challenge, NDPK-D–dependent TRAF6 mitochondrial recruitment triggers an energetic fitness checkpoint required to engage and maintain the transcriptional program necessary for inflammasome activation.

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The Blood Coagulation Cascade in a Perfusion Experiment: Example from the Pharmaceutical Industry

Evolution Equations Arising in the Modelling of Life Sciences ◽

10.1007/978-3-0348-0615-2_6 ◽

2012 ◽

pp. 195-207

Author(s):

Messoud Efendiev

Keyword(s):

Pharmaceutical Industry ◽

Blood Coagulation ◽

Coagulation Cascade ◽

Perfusion Experiment

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343 Tissue factor expressed on monocytes induces inter-cellular gap formation of vascular endothelial cells via the extrinsic blood coagulation cascade

Journal of Investigative Dermatology ◽

10.1016/j.jid.2019.07.345 ◽

2019 ◽

Vol 139 (9) ◽

pp. S273

Author(s):

R. Saito ◽

Y. Yanase ◽

T. Kawaguchi ◽

K. Uchida ◽

M. Hide

Keyword(s):

Endothelial Cells ◽

Tissue Factor ◽

Blood Coagulation ◽

Vascular Endothelial Cells ◽

Coagulation Cascade ◽

Vascular Endothelial ◽

Gap Formation

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Bioactive Lipids Metabolites in Amanita Virosa

Natural Product Communications ◽

10.1177/1934578x1200701121 ◽

2012 ◽

Vol 7 (11) ◽

pp. 1934578X1200701 ◽

Cited By ~ 1

Author(s):

Francesca Cateni ◽

Marina Zacchigna ◽

Bojan Doljak ◽

Marko Anderluh ◽

Giuseppe Procida ◽

...

Keyword(s):

Fatty Acids ◽

Free Fatty Acids ◽

Blood Coagulation ◽

Inhibitory Activity ◽

Complex Mixture ◽

Serine Proteinase ◽

Coagulation Cascade ◽

Bioactive Lipids ◽

Thrombin Inhibition ◽

Suitable Target

Thrombin is the key serine proteinase of the coagulation cascade and, therefore, a suitable target for inhibition of blood coagulation. An extract of Amanita virosa considerably inhibited thrombin (48%), but showed no inhibitory activity on trypsin. On the basis of inhibition selectivity between thrombin and trypsin and potency of thrombin inhibition, A. virosa constitutes a good starting material for the isolation of further compounds that are active against thrombin. Bioassay oriented fractionation of the extract of A. virosa led to the isolation of a complex mixture of triglycerides (TGs), monoacylglycerols (MAGs), free fatty acids (FAs) and ergosterol. The structures of the isolated lipids metabolites were determined on the basis of chemical and spectroscopic evidences.

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