Pre-Clinical Model to Study Recurrent Venous Thrombosis in the Inferior Vena Cava

Elizabeth A. Andraska; Catherine E. Luke; Megan A. Elfline; Samuel P. Henke; Siddharth S. Madapoosi; Allan K. Metz; Megan E. Hoinville; Thomas W. Wakefield; Peter K. Henke; Jose A. Diaz

doi:10.1055/s-0038-1645855

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 2018; 118(06): 1048-1057
DOI: 10.1055/s-0038-1645855

Stroke, Systemic or Venous Thromboembolism

Schattauer GmbH Stuttgart

Pre-Clinical Model to Study Recurrent Venous Thrombosis in the Inferior Vena Cava

Elizabeth A. Andraska

¹Department of Surgery, Section of Vascular Surgery, Conrad Jobst Vascular Research Laboratories, University of Michigan, Ann Arbor, Michigan, United States

,

Catherine E. Luke

¹Department of Surgery, Section of Vascular Surgery, Conrad Jobst Vascular Research Laboratories, University of Michigan, Ann Arbor, Michigan, United States

,

Megan A. Elfline

¹Department of Surgery, Section of Vascular Surgery, Conrad Jobst Vascular Research Laboratories, University of Michigan, Ann Arbor, Michigan, United States

,

Samuel P. Henke

¹Department of Surgery, Section of Vascular Surgery, Conrad Jobst Vascular Research Laboratories, University of Michigan, Ann Arbor, Michigan, United States

,

Siddharth S. Madapoosi

¹Department of Surgery, Section of Vascular Surgery, Conrad Jobst Vascular Research Laboratories, University of Michigan, Ann Arbor, Michigan, United States

,

Allan K. Metz

¹Department of Surgery, Section of Vascular Surgery, Conrad Jobst Vascular Research Laboratories, University of Michigan, Ann Arbor, Michigan, United States

,

Megan E. Hoinville

¹Department of Surgery, Section of Vascular Surgery, Conrad Jobst Vascular Research Laboratories, University of Michigan, Ann Arbor, Michigan, United States

,

Thomas W. Wakefield

¹Department of Surgery, Section of Vascular Surgery, Conrad Jobst Vascular Research Laboratories, University of Michigan, Ann Arbor, Michigan, United States

,

Peter K. Henke

¹Department of Surgery, Section of Vascular Surgery, Conrad Jobst Vascular Research Laboratories, University of Michigan, Ann Arbor, Michigan, United States

,

Jose A. Diaz

¹Department of Surgery, Section of Vascular Surgery, Conrad Jobst Vascular Research Laboratories, University of Michigan, Ann Arbor, Michigan, United States

› Author Affiliations

Abstract

Background Patients undergoing deep vein thrombosis (VT) have over 30% recurrence, directly increasing their risk of post-thrombotic syndrome. Current murine models of inferior vena cava (IVC) VT model host one thrombosis event.

Objective We aimed to develop a murine model to study IVC recurrent VT in mice.

Materials and Methods An initial VT was induced using the electrolytic IVC model (EIM) with constant blood flow. This approach takes advantage of the restored vein lumen 21 days after a single VT event in the EIM demonstrated by ultrasound. We then induced a second VT 21 days later, using either EIM or an IVC ligation model for comparison. The control groups were a sham surgery and, 21 days later, either EIM or IVC ligation. IVC wall and thrombus were harvested 2 days after the second insult and analysed for IVC and thrombus size, gene expression of fibrotic markers, histology for collagen and Western blot for citrullinated histone 3 (Cit-H3) and fibrin.

Results Ultrasound confirmed the first VT and its progressive resolution with an anatomical channel allowing room for the second thrombus by day 21. As compared with a primary VT, recurrent VT has heavier walls with significant up-regulation of transforming growth factor-β (TGF-β), elastin, interleukin (IL)-6, matrix metallopeptidase 9 (MMP9), MMP2 and a thrombus with high citrullinated histone-3 and fibrin content.

Conclusion Experimental recurrent thrombi are structurally and compositionally different from the primary VT, with a greater pro-fibrotic remodelling vein wall profile. This work provides a VT recurrence IVC model that will help to improve the current understanding of the biological mechanisms and directed treatment of recurrent VT.

Keywords

animal model - disease models - recurrence - venous thrombosis - venous thromboembolism

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References

References
1 Heit JA. The epidemiology of venous thromboembolism in the community. Arterioscler Thromb Vasc Biol 2008; 28 (03) 370-372
2 Schulman S, Lindmarker P, Holmström M. , et al. Post-thrombotic syndrome, recurrence, and death 10 years after the first episode of venous thromboembolism treated with warfarin for 6 weeks or 6 months. J Thromb Haemost 2006; 4 (04) 734-742
3 Henke PK, Comerota AJ. An update on etiology, prevention, and therapy of postthrombotic syndrome. J Vasc Surg 2011; 53 (02) 500-509
4 Kahn SR, Shbaklo H, Lamping DL. , et al. Determinants of health-related quality of life during the 2 years following deep vein thrombosis. J Thromb Haemost 2008; 6 (07) 1105-1112
5 Kahn SR, Shapiro S, Wells PS. , et al; SOX trial investigators. Compression stockings to prevent post-thrombotic syndrome: a randomised placebo-controlled trial. Lancet 2014; 383 (9920): 880-888
6 Enden T, Haig Y, Kløw NE. , et al; CaVenT Study Group. Long-term outcome after additional catheter-directed thrombolysis versus standard treatment for acute iliofemoral deep vein thrombosis (the CaVenT study): a randomised controlled trial. Lancet 2012; 379 (9810): 31-38
7 Kahn SR. The post-thrombotic syndrome: progress and pitfalls. Br J Haematol 2006; 134 (04) 357-365
8 Delis KT, Bountouroglou D, Mansfield AO. Venous claudication in iliofemoral thrombosis: long-term effects on venous hemodynamics, clinical status, and quality of life. Ann Surg 2004; 239 (01) 118-126
9 Hara T, Truelove J, Tawakol A. , et al. 18F-fluorodeoxyglucose positron emission tomography/computed tomography enables the detection of recurrent same-site deep vein thrombosis by illuminating recently formed, neutrophil-rich thrombus. Circulation 2014; 130 (13) 1044-1052
10 Diaz JA, Hawley AE, Alvarado CM. , et al. Thrombogenesis with continuous blood flow in the inferior vena cava. A novel mouse model. Thromb Haemost 2010; 104 (02) 366-375
11 Diaz JA, Wrobleski SK, Hawley AE, Lucchesi BR, Wakefield TW, Myers Jr DD. Electrolytic inferior vena cava model (EIM) of venous thrombosis. J Vis Exp 2011; (53) e2737
12 Diaz JA, Alvarado CM, Wrobleski SK. , et al. The electrolytic inferior vena cava model (EIM) to study thrombogenesis and thrombus resolution with continuous blood flow in the mouse. Thromb Haemost 2013; 109 (06) 1158-1169
13 Diaz JA, Obi AT, Myers Jr DD. , et al. Critical review of mouse models of venous thrombosis. Arterioscler Thromb Vasc Biol 2012; 32 (03) 556-562
14 Pamer OR, Shaydakov ME, Rainey JP, Lawrence DA, Greve JM, Diaz JA. Update on the electrolytic IVC model for pre-clinical studies of venous thrombosis. Res Pract Thromb Haemost 2018; 2 (02) 1-8
15 Wrobleski SK, Farris DM, Diaz JA, Myers Jr DD, Wakefield TW. Mouse complete stasis model of inferior vena cava thrombosis. J Vis Exp 2011; 52 (52) 2738
16 Deatrick KB, Obi A, Luke CE. , et al. Matrix metalloproteinase-9 deletion is associated with decreased mid-term vein wall fibrosis in experimental stasis DVT. Thromb Res 2013; 132 (03) 360-366
17 Kähäri VM, Olsen DR, Rhudy RW, Carrillo P, Chen YQ, Uitto J. Transforming growth factor-beta up-regulates elastin gene expression in human skin fibroblasts. Evidence for post-transcriptional modulation. Lab Invest 1992; 66 (05) 580-588
18 Hinz B, Celetta G, Tomasek JJ, Gabbiani G, Chaponnier C. Alpha-smooth muscle actin expression upregulates fibroblast contractile activity. Mol Biol Cell 2001; 12 (09) 2730-2741
19 DeRoo EP, Wrobleski SK, Shea EM. , et al. The role of galectin-3 and galectin-3-binding protein in venous thrombosis. Blood 2015; 125 (11) 1813-1821
20 Wojcik BM, Wrobleski SK, Hawley AE, Wakefield TW, Myers Jr DD, Diaz JA. Interleukin-6: a potential target for post-thrombotic syndrome. Ann Vasc Surg 2011; 25 (02) 229-239
21 Fuchs TA, Brill A, Wagner DD. Neutrophil extracellular trap (NET) impact on deep vein thrombosis. Arterioscler Thromb Vasc Biol 2012; 32 (08) 1777-1783
22 Diaz JA, Fuchs TA, Jackson TO. , et al; for the Michigan Research Venous Group*. Plasma DNA is elevated in patients with deep vein thrombosis. J Vasc Surg Venous Lymphat Disord 2013; 1(4)
23 Diaz JA, Wrobleski SK, Alvarado CM. , et al. P-selectin inhibition therapeutically promotes thrombus resolution and prevents vein wall fibrosis better than enoxaparin and an inhibitor to von Willebrand factor. Arterioscler Thromb Vasc Biol 2015; 35 (04) 829-837
24 Hara T, Bhayana B, Thompson B. , et al. Molecular imaging of fibrin deposition in deep vein thrombosis using fibrin-targeted near-infrared fluorescence. JACC Cardiovasc Imaging 2012; 5 (06) 607-615