scholarly journals Effects of Plasminogen Activator Inhibitor-1–Specific RNA Aptamers on Cell Adhesion, Motility, and Tube Formation

2011 ◽  
Vol 21 (6) ◽  
pp. 373-381 ◽  
Author(s):  
Stephanie Brandal ◽  
Charlene M. Blake ◽  
Bruce A. Sullenger ◽  
Yolanda M. Fortenberry
Keyword(s):  
Cell Adhesion ◽  
Plasminogen Activator ◽  
Plasminogen Activator Inhibitor ◽  
Tube Formation ◽  
Rna Aptamers ◽  
Plasminogen Activator Inhibitor 1 ◽  
Activator Inhibitor

scholarly journals The active conformation of plasminogen activator inhibitor 1, a target for drugs to control fibrinolysis and cell adhesion

Structure ◽  
1999 ◽  
Vol 7 (2) ◽  
pp. 111-118 ◽  
Author(s):  
Allan M Sharp ◽  
Penelope E Stein ◽  
Navraj S Pannu ◽  
Robin W Carrell ◽  
Mitchell B Berkenpas ◽  
...  
Keyword(s):  
Cell Adhesion ◽  
Plasminogen Activator ◽  
Plasminogen Activator Inhibitor ◽  
Active Conformation ◽  
Plasminogen Activator Inhibitor 1 ◽  
Activator Inhibitor

Inactivation of Plasminogen Activator Inhibitor-1 by RNA Aptamer Molecules

Blood ◽  
2012 ◽  
Vol 120 (21) ◽  
pp. 1107-1107
Author(s):  
Yolanda Fortenberry ◽  
Jared Damare
Keyword(s):  
Plasminogen Activator ◽  
Active Site ◽  
Plasminogen Activator Inhibitor ◽  
Rna Aptamers ◽  
Plasminogen Activator Inhibitor 1 ◽  
Rna Molecules ◽  
Type Plasminogen Activator ◽  
Antiproteolytic Activity ◽  
Activator Inhibitor ◽  
Pai 1

Abstract Abstract 1107 Introduction: The serine protease inhibitor (serpin) plasminogen activator inhibitor-1 (PAI-1), binds and inhibits the following plasminogen activators: tissue-type plasminogen activator (tPA), and urokinase-type plasminogen activator (uPA). This decreases plasmin production and triggers the dissolution of fibrin clots. Elevated levels of PAI-1 have been correlated with an increased risk for cardiovascular disease, as well as obesity and metabolic syndrome. Consequently, pharmacologically suppressing PAI-1 might prevent, or successfully treat vascular disease. Several PAI-1 small molecule inhibitors have recently been studied (PAI-039 is the best characterized). Since PAI-1 is a multifunctional protein, completely inhibiting PAI-1 may hinder its other functions. Therefore, it is important to independently develop inhibitors to the various regions of PAI-1. This can be accomplished by using small RNA molecules (aptamers) that bind with high affinity and specificity to individual protein domains. We recently published a paper showing how PAI-1 specific RNA aptamers bind to the heparin/vitronectin binding site of PAI-1 (Blake et al., 2009). We demonstrated that PAI-1 specific aptamers prevent cancer cells from detaching from vitronectin (in the presence of PAI-1), resulting in increased cell adhesion. These aptamers had no effect on PAI-1's other functions, particularly its antiproteolytic activity. Objective: This study's goal was to develop RNA aptamers to the active site of PAI-1; thereby, preventing the ability of PAI-1 to interact with plasminogen activators (tPA and uPA). Methods: The aptamers were generated by the systematic evolution of ligands by exponential enrichment (SELEX). Adopting the SELEX in vitro selection technique ensures the creation of nuclease-resistant RNA molecules that will bind to target proteins. We used in vitroassays to determine the effect of the aptamers on the interaction of PAI-1 with both tPA and uPA. Results: We isolated a family of aptamers that bind to wild-type PAI-1 with affinities in the nanomolar range. From this family, two aptamer clones (10–2 and 10–4) exhibited reduced binding to elastase cleaved PAI-1 and the PAI-1/tPA complex. This suggests that they bind to, or in the vicinity of, the active site. Using a chromogenic assay, we showed that the aptamer clone 10–4, and (to a lesser extent) the aptamer clone 10–2, inhibited PAI-1's antiproteolytic activity against tPA, further suggesting that these clones bind to PAI-1 within its active site region. Interestingly, neither clone was able to prevent PAI-1 from inhibiting uPA activity. Both aptamer clones disrupted PAI-1's ability to form a stable covalent complex with tPA. Increasing aptamer concentrations positively correlated with an increase in cleaved PAI-1, suggesting that these aptamer clones convert PAI-1 from an inhibitor to a substrate. Furthermore, we showed that both aptamer clones are able to inhibit PAI-1's activity in the presence of vitronectin. Conclusions: We have shown that we are able to inhibit one of PAI-1's functions without hindering its other functions. To our knowledge, this is the first report of an RNA molecule that is able to inhibit the antiproteolytic activity of PAI-1. We have generated two specific RNA aptamer molecules that hinder the ability of PAI-1 to interact with tPA, which has the potential to be used as an antithrombotic agent. Disclosures: No relevant conflicts of interest to declare.

The plasminogen activator inhibitor-1 gene is induced by cell adhesion through the MEK/ERK pathway

2003 ◽  
Vol 10 (6) ◽  
pp. 738-745 ◽  
Author(s):  
Hang Chang ◽  
Kou-Gi Shyu ◽  
Shankung Lin ◽  
Shiow-Chwen Tsai ◽  
Bao-Wei Wang ◽  
...  
Keyword(s):  
Cell Adhesion ◽  
Plasminogen Activator ◽  
Plasminogen Activator Inhibitor ◽  
Erk Pathway ◽  
Plasminogen Activator Inhibitor 1 ◽  
Activator Inhibitor

scholarly journals Defining the Proinflammatory Phenotype Using High Sensitive C-Reactive Protein Levels as the Biomarker

2005 ◽  
Vol 90 (8) ◽  
pp. 4549-4554 ◽  
Author(s):  
Sridevi Devaraj ◽  
Grant O’Keefe ◽  
Ishwarlal Jialal
Keyword(s):  
Cell Adhesion ◽  
Plasminogen Activator ◽  
Body Mass ◽  
Whole Blood ◽  
Plasminogen Activator Inhibitor ◽  
C Reactive Protein ◽  
Plasminogen Activator Inhibitor 1 ◽  
Reactive Protein ◽  
Tnf Α ◽  
Activator Inhibitor

Context: Inflammation is pivotal in atherosclerosis. The prototypic marker of inflammation is C-reactive protein (CRP). Numerous studies have confirmed that high CRP levels in normal volunteers predict cardiovascular events. Objective: The objective of this study was to define proximal and associated abnormalities of the proinflammatory phenotype using CRP levels as the biomarker. Design and Subjects: Two groups of normal, healthy subjects, selected by stringent criteria from an initial cohort of 252, were studied over the period of 12 months. Group 1 included subjects with consistently low CRP (<0.004 μm or <0.5 mg/liter; low CRP group; n = 15). Group 2 included subjects with consistently high CRP (>2.0 or >0.016 μm to <10 mg/liter or <0.085 μm; high CRP group; n = 13). Main Outcome Measures: Fasting blood (50 ml) was obtained, and the following parameters were assayed: high sensitivity CRP, fibrinogen, lipid profile, insulin, whole blood cytokines after stimulation with lipopolysaccharide (LPS; 100 ng/ml for 24 h), soluble cell adhesion molecules, plasminogen activator inhibitor-1, CD40, CD40 ligand, leptin, adiponectin, monocyte chemoattractant protein-1, IL-8, matrix metalloproteinase-3 (MMP-3), and MMP-9. Genomic DNA was obtained from peripheral blood leukocytes, and the TNF-α −308 genotype was determined. Results: The median CRP levels were 0.0018 μm (0.21 mg/liter) and 0.031 μm (3.7 mg/liter) for the low and high groups, respectively. High CRP subjects were older and had significantly higher body mass indexes, triglycerides, insulin, homeostasis model assessment, and leptin levels compared with low CRP subjects. The markers of inflammation, plasminogen activator inhibitor-1, MMP-9, fibrinogen, and vascular cell adhesion molecule-1 levels were significantly higher in the high compared with the low CRP group. LPS-stimulated levels of whole blood IL-1β, IL-6, and TNF were significantly higher, and IL-4 levels were significantly lower in the high CRP group. After age- and body mass index-adjusted analysis of covariance, only plasma MMP-9 levels and LPS-stimulated whole blood IL-1β and TNF levels were significantly higher in the high CRP group. The frequency of the rare A allele at TNF-α −308 was equivalent in high and low CRP groups. Conclusions: A phenotype characterized by increased plasma inflammatory mediators as well as increased LPS-stimulated whole blood TNF-α and IL-1β levels is associated with high plasma CRP levels. This systemic inflammatory phenotype may contribute to vascular inflammation or may reflect inflammation in vessels or at other sites.

RNA Aptamers as Conformational Probes and Regulatory Agents for Plasminogen Activator Inhibitor-1

Biochemistry ◽  
2010 ◽  
Vol 49 (19) ◽  
pp. 4103-4115 ◽  
Author(s):  
Jeppe B. Madsen ◽  
Daniel M. Dupont ◽  
Thomas B. Andersen ◽  
Anne F. Nielsen ◽  
Lu Sang ◽  
...  
Keyword(s):  
Plasminogen Activator ◽  
Plasminogen Activator Inhibitor ◽  
Rna Aptamers ◽  
Plasminogen Activator Inhibitor 1 ◽  
Activator Inhibitor
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