Locating the active site for angiogenesis and cell proliferation due to fibrin fragment E with a phage epitope display library

Stirk, C.M.; Reid, Alexander; Melvin, William T.; Thompson, W. D.

doi:10.1016/s0306-3623(01)00114-8

Cited by 10 publications

(11 citation statements)

References 20 publications

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“…In the context of our experimental approach, endothelial cell tube formation has been reported to be dependent on plasmin activity [45], [46]. Moreover, fibrin degradation products generated from fibrin cleaved by plasmin can also induce angiogenesis [47], [48]. In the current study, the dependence of apo(a) inhibition of tube formation in HUVECs on its ability to interfere with the plasminogen activation system was demonstrated by ‘activating’ (adding plasmin or uPA) or ‘deactivating’ (removing plasminogen from the serum in the medium) the plasminogen system.…”

Section: Discussionmentioning

confidence: 90%

Apolipoprotein(a) Inhibits In Vitro Tube Formation in Endothelial Cells: Identification of Roles for Kringle V and the Plasminogen Activation System

2013

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Elevated plasma concentrations of lipoprotein(a) are associated with increased risk for atherothrombotic diseases. Apolipoprotein(a), the unique glycoprotein component of lipoprotein(a), is characterized by the presence of multiple kringle domains, and shares a high degree of sequence homology with the serine protease zymogen plasminogen. It has been shown that angiostatin, a proteolytic fragment of plasminogen containing kringles 1–4, can effectively inhibit angiogenesis. Moreover, proteolytic fragments of plasminogen containing kringle 5 are even more potent inhibitors of angiogenesis than angiostatin. Despite its strong similarity with plasminogen, the role of apolipoprotein(a) in angiogenesis remains controversial, with both pro- and anti-angiogenic effects reported. In the current study, we evaluated the ability of apolipoprotein(a) to inhibit VEGF- and angiopoietin-induced tube formation in human umbilical cord endothelial cells. A 17 kringle-containing form of recombinant apo(a) (17K), corresponding to a well-characterized, physiologically-relevant form of the molecule, effectively inhibited tube formation induced by either VEGF or angiopoietin-1. Using additional recombinant apolipoprotein(a) (r-apo(a)) variants, we demonstrated that this effect was dependent on the presence of an intact lysine-binding site in kringle V domain of apo(a), but not on the presence of the functional lysine-binding site in apo(a) kringle IV type 10; sequences within in the amino-terminal half of the molecule were also not required for the inhibitory effects of apo(a). We also showed that the apo(a)-mediated inhibition tube formation could be reversed, in part by the addition of plasmin or urokinase plasminogen activator, or by removal of plasminogen from the system. Further, we demonstrated that apo(a) treated with glycosidases to remove sialic acid was significantly less effective in inhibiting tube formation. This is the first report of a functional role for the glycosylation of apo(a) although the mechanisms underlying this observation remain to be determined in the context of angiogenesis.

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Section: Discussionmentioning

confidence: 90%

Apolipoprotein(a) Inhibits In Vitro Tube Formation in Endothelial Cells: Identification of Roles for Kringle V and the Plasminogen Activation System

2013

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“…12,13 Further angiogenic factors, such as the fibrin degradation product termed fragment E, 13,14 are abundant in wounds and form the major component of experimental wound extracts. [15][16][17] We have located the active site on fibrin E, 18 but the presumptive receptor is yet to be discovered. Both are potential targets for therapeutic intervention.…”

Section: Discussionmentioning

confidence: 99%

Prognosis in human melanoma: PAR‐1 expression is superior to other coagulation components and VEGF

Depasquale¹,

Thompson

2008

Histopathology

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It appears paradoxical that VEGF expression is not more predictive of recurrence, but even low expression may be sufficient for tumour angiogenesis and other factors must govern tumour aggression. Antagonism of VEGF may still prove a successful adjunct in future therapeutic trials. Both MVD and PAR-1 can be used as adjuncts to Breslow thickness and ulceration as prognostic indicators for melanoma, as they appear to give independent information for all thicknesses. PAR-1 expression is the best antibody marker of recurrence risk from those studied. It remains to be seen whether this methodology can predict response to novel antiangiogenic therapies currently entering trial.

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“…24 The peroxidase-ABC detection method was used for both antibodies. Before incubation with primary antibody, antigen retrieval was performed: chymotrypsin digestion (250 mg chymotrypsin and 250 mg CaCl 2 in 500 mL of PBS for 30 minutes) was required for CD42c and pressure cooking (microwavable pressure cooker containing 1L Tris buffer heated unsealed for 20 minutes at full power, then sealed to reach pressure for 7 minutes) for fibrin.…”

Section: Immunohistochemical Stainingmentioning

confidence: 99%

Thrombosis and Neointima Formation in Vein Grafts Are Inhibited by Locally Applied Aspirin Through Endothelial Protection

Torsney

Mayr

Zou

et al. 2004

Circulation Research

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Abstract-Vein graft failure within the first month after bypass surgery is largely because of thrombosis. However, systemic study of thrombus formation in vein grafts is still lacking, and few effective techniques are available to prevent this event. Herein, we analyzed the kinetics of thrombosis and tested the effectiveness of locally applied aspirin on prevention of the disease in a mouse model. En face analysis of vein grafts revealed that 67Ϯ12% and 54Ϯ17% of the surface areas were covered by microthrombi at 1 and 3 days, respectively. Thrombus generation was also identified by labeling of platelets and fibrin, which occurred in 35 grafts examined at 1 and 3 days and 1, 2, 4, and 8 weeks. In a fifth of grafts, the thrombus occluded the vessel lumen by Ն1/4. Furthermore, a significant loss of endothelial cells was evidenced by ␤-gal staining for vein grafts in transgenic mice expressing LacZ gene controlled by TIE2-endothelial specific gene promoter. Following thrombosis, neointimal lesions were significantly increased by 4-fold 2 weeks after the operation. When vein grafts were treated locally with aspirin in pluronic gel-127, the thrombus area was significantly reduced (PϽ0.005) at 1, 4, and 8 weeks. Interestingly, neointimal lesions were markedly reduced in the local, but not oral, aspirin-treated group at 4 and 8 weeks by 50% to 70% (PϽ0.005). The mechanism of reduced lesions by locally applied aspirin involved the protection of vein graft endothelium. Thus, we provide strong evidence that thrombus formation occurs before the development of neointimal lesions in vein grafts and that local aspirin treatment successfully reduces vein graft arteriosclerosis through endothelial protection, resulting in reduction of thrombosis.

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Locating the active site for angiogenesis and cell proliferation due to fibrin fragment E with a phage epitope display library

Cited by 10 publications

References 20 publications

Apolipoprotein(a) Inhibits In Vitro Tube Formation in Endothelial Cells: Identification of Roles for Kringle V and the Plasminogen Activation System

Apolipoprotein(a) Inhibits In Vitro Tube Formation in Endothelial Cells: Identification of Roles for Kringle V and the Plasminogen Activation System

Prognosis in human melanoma: PAR‐1 expression is superior to other coagulation components and VEGF

Thrombosis and Neointima Formation in Vein Grafts Are Inhibited by Locally Applied Aspirin Through Endothelial Protection

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