Role of coagulation factor XIII in fibrin clot structure and thrombosis

Lead Research Organisation: University of Leeds
Department Name: School of Medicine

Abstract

Clot formation within a blood vessel is the predominant cause of heart attacks, stroke and deep vein thrombosis. The clot is composed of a protein called fibrin which undergoes cross-linking by an enzyme called transglutaminase (factor XIII). The role of factor XIII in clot structure and thrombosis is not well understood. Our research is aimed at discovering how the thrombus is modulated by factor XIII, using model systems of clot formation, engineered proteins and in vivo models. This study will develop better understanding of processes involved in clot stability and in the future will help to design new anti-clotting drugs.

Technical Summary

The thrombus is composed of a mesh of fibrin fibres, with platelets, erythrocytes and other cells embedded in this network. The clot is stabilised by cross-linking activity of transglutaminase FXIIIa. The effect(s) of cross-linking on fibrin clot structure and on risk for thrombosis are unknown. The aims of this project are to characterise 1) effects of cross-linking on fibrin structure and mechanisms by which effects occurs, 2) the relationship between fibrin cross-linking and FXIII activation, and 3) the contribution of cross-linking to in vivo thrombus formation. Understanding the mechanisms that regulate clot stability will facilitate discovery of new treatments for thrombosis.

Publications

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Ariëns RA (2013) Counting 1 fibrin molecule at a time. in Blood

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Ariëns RA (2013) Fibrin(ogen) and thrombotic disease. in Journal of thrombosis and haemostasis : JTH

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Ariëns RA (2011) Elevated fibrinogen causes thrombosis. in Blood

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Bridge KI (2014) Clot properties and cardiovascular disease. in Thrombosis and haemostasis

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Casini A (2015) Dysfibrinogenemia: from molecular anomalies to clinical manifestations and management. in Journal of thrombosis and haemostasis : JTH

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Cilia La Corte AL (2011) Role of fibrin structure in thrombosis and vascular disease. in Advances in protein chemistry and structural biology

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Duval C (2017) Fibrinogen splice variation and cross-linking: Effects on fibrin structure/function and role of fibrinogen ?' as thrombomobulin II. in Matrix biology : journal of the International Society for Matrix Biology

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Duval C (2016) Factor XIII A-Subunit V34L Variant Affects Thrombus Cross-Linking in a Murine Model of Thrombosis. in Arteriosclerosis, thrombosis, and vascular biology

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Hethershaw EL (2014) The effect of blood coagulation factor XIII on fibrin clot structure and fibrinolysis. in Journal of thrombosis and haemostasis : JTH

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Kohler HP (2011) Diagnosis and classification of factor XIII deficiencies. in Journal of thrombosis and haemostasis : JTH

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Scott DJ (2011) Clot architecture is altered in abdominal aortic aneurysms and correlates with aneurysm size. in Arteriosclerosis, thrombosis, and vascular biology

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Smith EL (2013) Elimination of coagulation factor XIII from fibrinogen preparations. in Journal of thrombosis and haemostasis : JTH

 
Description Official ISTH SSC publication on FXIII deficiency
Geographic Reach Multiple continents/international 
Policy Influence Type Implementation circular/rapid advice/letter to e.g. Ministry of Health
 
Description Official ISTH SSC publication on standard clot permeation method
Geographic Reach Multiple continents/international 
Policy Influence Type Implementation circular/rapid advice/letter to e.g. Ministry of Health
 
Description Genome Editing Mice For Medicine (GEMM) Initiative
Amount £30,000 (GBP)
Organisation Research Complex at Harwell 
Sector Public
Country United Kingdom
Start 10/2016 
End 09/2017
 
Description Programme Grant
Amount £1,145,224 (GBP)
Funding ID RG/13/3/30104 
Organisation British Heart Foundation (BHF) 
Sector Charity/Non Profit
Country United Kingdom
Start 10/2013 
End 09/2018
 
Title AFM study of fibrin polymerisation 
Description We study structural and kynetic processes involved fibrin polymerisation at sub-molecular level using atomic force microscopy (AFM). 
Type Of Material Model of mechanisms or symptoms - in vitro 
Year Produced 2013 
Provided To Others? Yes  
Impact We are currently the only laboratory in the world studying fibrin polymerisation at high molecular resolution using the AFM. We have interest from several peers (Aberdeen and Geneva) who wish to collaborate and learn this methodology. 
 
Title In vivo thrombosis model 
Description We have developed a murine FeCl3 and laser induced thrombosis model of thrombosis in the femoral vessels and cremaster arterioles. We also have factor XIII deficient and fibinogen deficient mice available for studies with recombinant FXIII and fibrinogen variants. 
Type Of Material Model of mechanisms or symptoms - mammalian in vivo 
Provided To Others? No  
Impact Our in vivo thrombosis model is unique in that we have developed excvellent quntification tools for kinetic analysis of thrombus growth, size and fibrinolysis rates. 
 
Title Laser scanning confocal imaging of fibrin polymerisation 
Description As part of the methods devloped for this grant, we have developed a powerful new method of Laser scanning confocal imaging of fibrin polymerisation. 
Type Of Material Technology assay or reagent 
Year Produced 2012 
Provided To Others? Yes  
Impact The new method of Laser scanning confocal imaging of fibrin polymerisation enables us to integrate fibrin polymerisation data from light scattering with morphological analysis of fibrin formation. 
 
Title Magnetic Tweezers 
Description Magnetic tweezers have been developed in our laboratory to study the visco-elastic properties of the fibrin clot. 
Type Of Material Model of mechanisms or symptoms - in vitro 
Provided To Others? No  
Impact We are the only laboratory currently using this methodology for fibrin clots worldwide. 
 
Title Recombinant FXIII expression 
Description We express recombinant human blood coagulation factor XIII and variants thereof in E Coli. 
Type Of Material Cell line 
Provided To Others? No  
Impact We are one of a few laboratories around the world currently expression recombinant human FXIII 
 
Title Recombinant fibrinogen expression 
Description We express recombinant human fibrinogen and variants thereof in CHO and BHK cells. 
Type Of Material Cell line 
Year Produced 2013 
Provided To Others? Yes  
Impact We are one of a few laboratories around the world currently expressing recombinant human fibrinogen. 
 
Description Alisa Wolberg 
Organisation University of North Carolina at Chapel Hill
Department Department of Pathology and Laboratory Medicine
Country United States 
Sector Academic/University 
PI Contribution Dr Cedric Duval and I contributed invaluable proteins and cells for the research, assisted in research design and interpreted data.
Collaborator Contribution We collaborate with the Wolberg lab by exchange of proteins and cells, which leads to joint publications in high-impact journals.
Impact Factor XIIIa-dependent retention of red blood cells in clots is mediated by fibrin a-chain crosslinking. Byrnes JR, Duval C, Wang Y, Hansen CE, Ahn B, Mooberry MJ, Clark MA, Johnsen JM, Lord ST, Lam WA, Meijers JC, Ni H, Ariëns RA, Wolberg AS. Blood. 2015 Oct 15;126(16):1940-8.
Start Year 2014
 
Description Elisabetta Castoldi 
Organisation Maastricht University (UM)
Country Netherlands 
Sector Academic/University 
PI Contribution My laboratory has provided invaluable research reagents (protein), assisted in study design, obtained joint grant income, and interpreted data.
Collaborator Contribution Development of new research techniques and new research area.
Impact Fibrinogen ?' increases the sensitivity to activated protein C in normal and factor V Leiden plasma. Omarova F, Uitte de Willige S, Simioni P, Ariëns RA, Bertina RM, Rosing J, Castoldi E. Blood. 2014 Aug 28;124(9):1531-8. doi: 10.1182/blood-2014-02-554055. Epub 2014 Jun 20. Inhibition of thrombin-mediated factor V activation contributes to the anticoagulant activity of fibrinogen ?'. Omarova F, Uitte De Willige S, Ariëns RA, Rosing J, Bertina RM, Castoldi E. J Thromb Haemost. 2013 Sep;11(9):1669-78. doi: 10.1111/jth.12354.
Start Year 2012
 
Description Hugo ten Cate 
Organisation Maastricht University (UM)
Country Netherlands 
Sector Academic/University 
PI Contribution The ten Cate laboratory and my laboratory exchange samples, reagents and researchers for several collaborative studies.
Collaborator Contribution The ten Cate laboratory and my laboratory exchange samples, reagents and researchers for several collaborative studies.
Impact Clot structure and fibrinolytic potential in patients with post thrombotic syndrome. Bouman AC, McPherson H, Cheung YW, Ten Wolde M, Ten Cate H, Ariëns RA, Ten Cate-Hoek AJ. Thromb Res. 2016 Jan;137:85-91. doi: 10.1016/j.thromres.2015.11.013. Epub 2015 Nov 10. The role of activated coagulation factor XII in overall clot stability and fibrinolysis. Konings J, Hoving LR, Ariëns RS, Hethershaw EL, Ninivaggi M, Hardy LJ, de Laat B, Ten Cate H, Philippou H, Govers-Riemslag JW. Thromb Res. 2015 Aug;136(2):474-80. doi: 10.1016/j.thromres.2015.06.028. Epub 2015 Jun 19. Factor XIIa regulates the structure of the fibrin clot independently of thrombin generation through direct interaction with fibrin. Konings J, Govers-Riemslag JW, Philippou H, Mutch NJ, Borissoff JI, Allan P, Mohan S, Tans G, Ten Cate H, Ariëns RA. Blood. 2011 Oct 6;118(14):3942-51. doi: 10.1182/blood-2011-03-339572.
Start Year 2010
 
Description FASEB 2013 Regulation of fibrin structure and its role in thrombosis 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Type Of Presentation Keynote/Invited Speaker
Geographic Reach International
Primary Audience Other academic audiences (collaborators, peers etc.)
Results and Impact This was an invited presentation at the 2013 FASEB meeting on proteases in haemostasis by Ariens.

The meeting allowed exposure of our most recent studies to international peer colleagues and USA colleagues in particular.
Year(s) Of Engagement Activity 2013
URL https://secure.faseb.org/FASEB/meetings/summrconf/Programs/11531.pdf
 
Description ISTH 2013 State-of-the-Art Lecture on the Role of Fibrin(ogen) in Thrombostic Disease 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Type Of Presentation Keynote/Invited Speaker
Geographic Reach International
Primary Audience Other academic audiences (collaborators, peers etc.)
Results and Impact The PI was invited to give a lecture on Fibrin(ogen) and Thrombotic Disease at the 2013 ISTH Congress in Amsterdam, June 29 - July 4

This was a state-of-the-art lecture at the main international conference on Thrombosis and Haemostasis (ISTH) by the PI. Ariens also wrote a manuscript and there were a few of oral communication sessions on the topic at the meeting.
Year(s) Of Engagement Activity 2013
 
Description On Blood Clotting and Thrombosis: The Clot Thickens 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? Yes
Geographic Reach National
Primary Audience Public/other audiences
Results and Impact Increased awareness of research group's activity

Invited to speak at other occasions
Year(s) Of Engagement Activity 2014
 
Description Press release and publication 
Form Of Engagement Activity A magazine, newsletter or online publication
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Media (as a channel to the public)
Results and Impact Our new BHF research programme award featured in the Yorkshire Post and Yorkshire Evening Post, July 2013.


Raising local public awareness of our research on clot structure.
Year(s) Of Engagement Activity 2013
 
Description Sky television interview 
Form Of Engagement Activity A press release, press conference or response to a media enquiry/interview
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Media (as a channel to the public)
Results and Impact The PI (Ariens) discussed thrombosis on live Sky Television (Chrissy B show, August 2013)

Raising awareness of thrombosis and clotting with the general public
Year(s) Of Engagement Activity 2013
URL http://www.youtube.com/watch?v=Y68kntUqkZY