Hemostasis & Thrombosis
The haemostasis and thrombosis research within the Department of Haematology is driven by the collaborative interests of Prof David Lane, Prof Mike Laffan, Dr Jim Crawley and Dr Carolyn Millar. Thrombosis, bleeding disorders and diseases of blood vessels are among the most prevalent causes of premature death in Western society and a major cause of morbidity. Advances in the basic science underlying the function of the enzymes, cofactors and inhibitory components of haemostasis are urgently needed to find ways of preventing the onset and progression of disease, and to improve the treatment of affected individuals.
Prof David Lane, Prof Mike Laffan, Dr Jim Crawley and Dr Carolyn Millar have all had long-standing interests in haemostatic mechanisms.
The major physiological anticoagulant pathways, involving tissue factor pathway inhibitor (TFPI), protein C and antithrombin pathways, are important for normal blood fluidity. Inherited deficiency of components of these pathways caused by gene defects can predispose individuals towards thrombosis.
Fig 1: Model highlighting different residues involved in mediating the anticoagulant and cytoprotective functions of APC
Previous studies have involved how protein C interacts with the endothelial cell protein C receptor (EPCR), and its cofactor, protein S, and also the study of the gene regulation of the EPCR, and how its expression is specifically directed to large vessel endothelial cells.
Current work includes projects that focus on the characterisation of the cytoprotective signalling function of APC. Activated protein C (APC) is a natural anticoagulant protein that in recent years has been found to transduce cytoprotective signals to endothelial cells. This signalling function has potentially major therapeutic implications for the treatment of patients with severe sepsis, or that have suffered from a stroke. Other anticoagulant research interests include structure/function studies on protein S and tissue factor pathway inhibitor (TFPI) and the physiological importance of their anticoagulant function.
The Haemostasis group also conducts research projects on von Willebrand factor (VWF) - a key protein that mediates platelet tethering at sites of vascular injury, and ADAMTS13 - a metalloprotease that cleaves VWF and thus modulates its function.
Fig 2: Model of ADAMTS13 metalloprotease domain highlighting the location of the active site (blue) that coordinates a Zn2+ ion, and residues involved in a high affinity Ca2+ binding site (purple) that influences ADAMTS13 function.
Abnormalities or deficiencies of these important proteins may result in thrombotic or haemorrhagic disorders such as thrombotic thrombocytopenic purpura and von Willebrand disease, respectively and are also risk factors for cardiovascular disease.
Current ADAMTS13 research activity includes structure/function studies that aim to delineate how ADAMTS13 interacts with and proteolyses VWF, how calcium modulates the activity of ADAMTS13, how the high degree of substrate specificity of ADAMTS13 is conferred, and how plasma ADAMTS13 levels/activity influence thrombotic risk.
Plasma levels of VWF are closely dependent on its numerous O- and N-linked sugar chains. Current research projects are exploring the functional consequences of N- and O-linked glycosylation on VWF conformation and on its many interactions with other molecules. Collaborative projects are producing a detailed glycan map of VWF and exploring the role of VWF in endothelial cell function.
Biographical sketch
Prof David Lane is a Professor of Molecular Haematology in the Department of Haematology. He obtained his PhD in 1974 from the University of London. He joined Imperial College (then Charing Cross and Westminster Medical School) in 1979. He is Editor-in-Chief of the Journal of Thrombosis & Haemostasis.
Prof Mike Laffan a Professor of Haemostasis and Thrombosis in the Department of Haematology. He qualified in medicine from Oxford University in 1981 and obtained his DM in 1993. Prof Mike Laffan joined Imperial College in 1992 and is also director of the Hammersmith Hospital Haemophilia Centre.
Dr Jim Crawley is a non-clinical lecturer. He graduated from Durham University in with a BSc in Molecular Biology and Biochemistry, and obtained his PhD from Imperial College London in 2001. He joined the Haematology Department in 2002. Dr Jim Crawley is Scientific Editor for the Journal of Thrombosis & Haemostasis.
Dr Carolyn Millar is a clinical senior lecturer. She joined Imperial College in 2008.
Haemostasis Group Members
Group Leaders
Prof David Lane, Prof Mike Laffan, Dr Jim Crawley, Dr Carolyn Millar
Clinical Lecturer
Dr James Uprichard
Post-doctoral Scientists
Dr (Alain) C Chion, Dr Brenda Luken, Dr Sara Zanardelli, Dr Tom McKinnon, Dr Ayesha Khan
PhD Students
Michelle Gardner, Rens De Groot, Maria Efthymiou, Verity Hockey, Agata Nowak, Susan Shapiro, Kevin Canis, Yaozu Xiang
Technician
(Jessica) Yu Yao
Visiting Scientist
Dr Jose Gonzalez Porras
Selected Publications
1 McKinnon TA, Chion AC, Millington AJ, Lane DA, Laffan MA. N-linked glycosylation of VWF modulates its interaction with ADAMTS13. Blood. 2008; 111: 3042-9.
2 Ghevaert C, Salsmann A, Watkins NA, Schaffner-Reckinger E, Rankin A, Garner SF, Stephens J, Smith GA, Debili N, Vainchenker W, de Groot PG, Huntington JA, Laffan M, Kieffer N, Ouwehand WH. A nonsynonymous SNP in the ITGB3 gene disrupts the conserved membrane-proximal cytoplasmic salt bridge in the alphaIIbbeta3 integrin and cosegregates dominantly with abnormal proplatelet formation and macrothrombocytopenia. Blood. 2008; 111: 3407-14.
3 Crawley JT, Lane DA. The haemostatic role of tissue factor pathway inhibitor. Arterioscler Thromb Vasc Biol. 2008; 28: 233-42.
4 Chion CK, Doggen CJ, Crawley JT, Lane DA, Rosendaal FR. ADAMTS13 and von Willebrand factor and the risk of myocardial infarction in men. Blood. 2007; 109: 1998-2000.
5 Zanardelli S, Crawley JT, Chion CK, Lam JK, Preston RJ, Lane DA. ADAMTS13 substrate recognition of von Willebrand factor A2 domain. J Biol Chem. 2006; 281: 1555-63.
6 Preston RJ, Ajzner E, Razzari C, Karageorgi S, Dua S, Dahlback B, Lane DA. Multifunctional specificity of the protein C/activated protein C Gla domain. J Biol Chem. 2006; 281: 28850-7.
7 Mollica LR, Crawley JT, Liu K, Rance JB, Cockerill PN, Follows GA, Landry JR, Wells DJ, Lane DA. Role of a 5'-enhancer in the transcriptional regulation of the human endothelial cell protein C receptor gene. Blood. 2006; 108: 1251-9.
8 O'Donnell JS, McKinnon TA, Crawley JT, Lane DA, Laffan MA. Bombay phenotype is associated with reduced plasma-VWF levels and an increased susceptibility to ADAMTS13 proteolysis. Blood. 2005; 106: 1988-91.
9 Lane DA, Philippou H, Huntington JA. Directing thrombin. Blood. 2005; 106: 2605-12.
10 Crawley JT, Lam JK, Rance JB, Mollica LR, O'Donnell JS, Lane DA. Proteolytic inactivation of ADAMTS13 by thrombin and plasmin. Blood. 2005; 105: 1085-93.
11 Rezende SM, Simmonds RE, Lane DA. Coagulation, inflammation, and apoptosis: different roles for protein S and the protein S-C4b binding protein complex. Blood. 2004; 103: 1192-201.
12 Hurtado V, Montes R, Gris JC, Bertolaccini ML, Alonso A, Martinez-Gonzalez MA, Khamashta MA, Fukudome K, Lane DA, Hermida J. Autoantibodies against EPCR are found in antiphospholipid syndrome and are a risk factor for fetal death. Blood. 2004; 104: 1369-74.
13 Rance JB, Follows GA, Cockerill PN, Bonifer C, Lane DA, Simmonds RE. Regulation of the human endothelial cell protein C receptor gene promoter by multiple Sp1 binding sites. Blood. 2003; 101: 4393-401.
14 Raja SM, Chhablani N, Swanson R, Thompson E, Laffan M, Lane DA, Olson ST. Deletion of P1 arginine in a novel antithrombin variant (antithrombin London) abolishes inhibitory activity but enhances heparin affinity and is associated with early onset thrombosis. J Biol Chem. 2003; 278: 13688-95.
15 Philippou H, Rance J, Myles T, Hall SW, Ariens RA, Grant PJ, Leung L, Lane DA. Roles of low specificity and cofactor interaction sites on thrombin during factor XIII activation. Competition for cofactor sites on thrombin determines its fate. J Biol Chem. 2003; 278: 32020-6.
16 Mille-Baker B, Rezende SM, Simmonds RE, Mason PJ, Lane DA, Laffan MA. Deletion or replacement of the second EGF-like domain of protein S results in loss of APC cofactor activity. Blood. 2003; 101: 1416-8.
17 Rezende SM, Lane DA, Mille-Baker B, Samama MM, Conard J, Simmonds RE. Protein S Gla-domain mutations causing impaired Ca(2+)-induced phospholipid binding and severe functional protein S deficiency. Blood. 2002; 100: 2812-9.
18 Ouyang YB, Crawley JT, Aston CE, Moore KL. Reduced body weight and increased postimplantation fetal death in tyrosylprotein sulfotransferase-1-deficient mice. J Biol Chem. 2002; 277: 23781-7.
19 O'Donnell J, Boulton FE, Manning RA, Laffan MA. Amount of H antigen expressed on circulating von Willebrand factor is modified by ABO blood group genotype and is a major determinant of plasma von Willebrand factor antigen levels. Arterioscler Thromb Vasc Biol. 2002; 22: 335-41.
20 Luttun A, Lupu F, Storkebaum E, Hoylaerts MF, Moons L, Crawley J, Bono F, Poole AR, Tipping P, Herbert JM, Collen D, Carmeliet P. Lack of plasminogen activator inhibitor-1 promotes growth and abnormal matrix remodeling of advanced atherosclerotic plaques in apolipoprotein E-deficient mice. Arterioscler Thromb Vasc Biol. 2002; 22: 499-505.
21 Kunz G, Ohlin AK, Adami A, Zoller B, Svensson P, Lane DA. Naturally occurring mutations in the thrombomodulin gene leading to impaired expression and function. Blood. 2002; 99: 3646-53.
22 Gu JM, Crawley JT, Ferrell G, Zhang F, Li W, Esmon NL, Esmon CT. Disruption of the endothelial cell protein C receptor gene in mice causes placental thrombosis and early embryonic lethality. J Biol Chem. 2002; 277: 43335-43.
23 Crawley JT, Goulding DA, Ferreira V, Severs NJ, Lupu F. Expression and localization of tissue factor pathway inhibitor-2 in normal and atherosclerotic human vessels. Arterioscler Thromb Vasc Biol. 2002; 22: 218-24.
24 Lane DA, Grant PJ. Role of hemostatic gene polymorphisms in venous and arterial thrombotic disease. Blood. 2000; 95: 1517-32.
25 Kunz G, Ireland HA, Stubbs PJ, Kahan M, Coulton GC, Lane DA. Identification and characterization of a thrombomodulin gene mutation coding for an elongated protein with reduced expression in a kindred with myocardial infarction. Blood. 2000; 95: 569-76.
26 Crawley J, Lupu F, Westmuckett AD, Severs NJ, Kakkar VV, Lupu C. Expression, localization, and activity of tissue factor pathway inhibitor in normal and atherosclerotic human vessels. Arterioscler Thromb Vasc Biol. 2000; 20: 1362-73.
27 Ariens RA, Philippou H, Nagaswami C, Weisel JW, Lane DA, Grant PJ. The factor XIII V34L polymorphism accelerates thrombin activation of factor XIII and affects cross-linked fibrin structure. Blood. 2000; 96: 988-95.
28 Al-Obaidi MK, Philippou H, Stubbs PJ, Adami A, Amersey R, Noble MM, Lane DA. Relationships between homocysteine, factor VIIa, and thrombin generation in acute coronary syndromes. Circulation. 2000; 101: 372-7.
29 Simmonds RE, Lane DA. Structural and functional implications of the intron/exon organization of the human endothelial cell protein C/activated protein C receptor (EPCR) gene: comparison with the structure of CD1/major histocompatibility complex alpha1 and alpha2 domains. Blood. 1999; 94: 632-41.
30 Bayston TA, Tripodi A, Mannucci PM, Thompson E, Ireland H, Fitches AC, Hananeia L, Olds RJ, Lane DA. Familial overexpression of beta antithrombin caused by an Asn135Thr substitution. Blood. 1999; 93: 4242-7.
