Reduction of early vein graft thrombosis by tissue plasminogen activator gene transfer
Anita C. Thomas
1
Bristol Heart Institute, University of Bristol, Bristol, United Kingdom
2
Australian Institute for Bioengineering and Nanotechnology, The University of Queensland, Brisbane, Australia
,
Marcella J. Wyatt
1
Bristol Heart Institute, University of Bristol, Bristol, United Kingdom
,
Andrew C. Newby
1
Bristol Heart Institute, University of Bristol, Bristol, United Kingdom
› InstitutsangabenFinancial support: ACT was the recipient of an Australian National Health and Medical Research Council Biomedical (CJ Martin) Overseas Fellowship (ID #252926). Additional support was obtained from the British Heart Foundation and the Medical Research Foundation.
Vein grafts are used to bypass coronary arterial stenosis, but many grafts thrombose soon after surgery. A model was developed in the pig to allow continuous measurement of blood flow and production of flow-restricting thrombi (cyclic flow reductions; CFRs). Saphenous vein lumen was exposed to adenovirus ex vivo, to over-express human tissue plasminogen activator (h-tPA), with β-galactosidase adenovirus as a control. The vein segmen ts were engrafted into carotid arteries and examined 0,1 or 3 days later (4–7 animals/group). Untransduced grafts examined on the day of surgery developed repeated CFRs at both normal and restricted flow, but their frequency declined in grafts examined after 3 days. Adenovirus transduction was evident as β-galactosidase or h-tPA expression 1 day after engraftment. Blood flow was increased 1.4-fold in h-tPA transduced grafts after 1 day [control 390 (280–510), h-tPA 550 (450–660) ml/min; p=0.02 (expressed as mean (95% confidence intervals)]. CFRs were less severe (p=0.002) in the h-tPA transduced grafts than β-galactosidase-transduced grafts. CFRs were also less frequent in unstenosed undamaged h-tPA grafts [control 17 (6.1–29), h-tPA 7.6 (1.7–14) CFR/hr; p=0.02], but this difference was reduced after damage or stenosis. CFRs formed faster in h-tPA than in β-galactosidase-transduced grafts [control 14 (11–17), h-tPA 23 (19–27) ml/min2; p<0.001], and resolved twofold faster [control 25 (22–30), h-tPA 48 (39–60) ml/min2; p<0.001]. Hence, in this model, local gene therapy with h-tPA increased graft blood flow and decreased measures of early graft thrombosis, namely quicker CFR resolution and decreased frequency and severity.
5
Underwood MJ,
Cooper G,
More R.
et al. Effect of intraluminal application of tissue-type plasminogen activator on the fibrinolytic activity of experimental vein grafts. Cardiovasc Res 1995; 29: 422-427.
6
Manchio JV,
Gu J,
Romar L.
et al. Disruption of graft endothelium correlates with early failure after offpump coronary artery bypass surgery. Ann Thorac Surg 2005; 79: 1991-1998.
7
Kauhanen P,
Siren V,
Carpen O.
et al. Plasminogen activator inhibitor-1 in neointima of vein grafts: its role in reduced fibrinolytic potential and graft failure. Circulation 1997; 96: 1783-1789.
8
Poston RS,
Gu J,
Brown JM.
et al. Endothelial injury and acquired aspirin resistance as promoters of regional thrombin formation and early vein graft failure after coronary artery bypass grafting. J Thorac Cardiovasc Surg 2006; 131: 122-130.
11
Strony J,
Beaudoin A,
Brands D.
et al. Analysis of shear stress and hemodynamic factors in a model of coronary artery stenosis and thrombosis. Am J Physiol 1993; 265: H1787-H1796.
12
Anderson HV,
Kirkeeide RL,
Krishnaswami A.
et al. Cyclic flow variations after coronary angioplasty in humans: clinical and angiographic characteristics and elimination with 7E3 monoclonal antiplatelet antibody. J Am Coll Cardiol 1994; 23: 1031-1037.
13
George SJ,
Lloyd CT,
Angelini GD.
et al. Inhibition of late vein graft neointima formation in human and porcine models by adenovirus-mediated overexpression of tissue inhibitor of metalloproteinase-3. Circulation 2000; 101: 296-304.
15
Carmeliet P,
Stassen JM,
van Vlaenderen I.
et al. Adenovirus-mediated transfer of tissue-type plasminogen activator augments thrombolysis in tissuetype plasminogen activator-deficient and plasminogen activator inhibitor-1-overexpressing mice. Blood 1997; 90: 1527-1534.
16
Wilkinson GW,
Akrigg A.
Constitutive and enhanced expression from the CMV major IE promoter in a defective adenovirus vector. Nucleic Acids Res 1992; 20: 2233-2239.
18
Golino P,
Cirillo P,
Calabro P.
et al. Expression of exogenous tissue factor pathway inhibitor in vivo suppresses thrombus formation in injured rabbit carotid arteries. J Am Coll Cardiol 2001; 38: 569-576.
19
Emeis JJ,
van den Eijnden-Schrauwen,
van den Hoogen CM.
et al. An endothelial storage granule for tissue-type plasminogen activator. J Cell Biol 1997; 139: 245-256.
20
Eckstein RW,
Gregg DE,
Pritchard W.
The magnitude and time of development of the collateral circulation in occluded femoral, carotid and coronary arteries. Am J Physiol 1941; 132: 351-361.
21
Waugh JM,
Kattash M,
Li J.
et al. Gene therapy to promote thromboresistance: local overexpression of tissue plasminogen activator to prevent arterial thrombosis in an in vivo rabbit model. Proc Natl Acad Sci USA 1999; 96: 1065-1070.
22
Kimura H,
Sakata Y,
Hamada H.
et al. In vivo retention of endothelial cells adenovirally transduced with tissue-type plasminogen activator and seeded onto expanded polytetrafluoroethylene. J Vasc Surg 2000; 32: 353-363.
25
Jiang X,
Liu X,
Zhang K.
et al. Experimental study of tissue-type plasminogen activator gene to prevent vein grafts stenosis. J Huazhong Univ Sci Technolog Med Sci 2006; 26: 314-316.
26
Burggraf D,
Martens HK,
Dichgans M.
et al. Matrix metalloproteinase (MMP) induction and inhibition at different doses of recombinant tissue plasminogen activator following experimental stroke. Thromb Haemost 2007; 98: 963-969.
27
Torsney E,
Mayr U,
Zou Y.
et al. Thrombosis and neointima formation in vein grafts are inhibited by locally applied aspirin through endothelial protection. Circ Res 2004; 94: 1466-1473.
29
Mutch NJ,
Moir E,
Robbie LA.
et al. Localization and identification of thrombin and plasminogen activator activities in model human thrombi by in situ zymography. Thromb Haemost 2002; 88: 996-1002.
30
Davies MG,
Klyachkin ML,
Dalen H.
et al. The integrity of experimental vein graft endothelium--implications on the etiology of early graft failure. Eur J Vasc Surg 1993; 07: 156-165.
31
Bourassa MG,
Campeau L,
Lesperance J.
et al. Changes in grafts and coronary arteries after saphenous vein aortocoronary bypass surgery: results at repeat angiography. Circulation 1982; 65: 90-97.
