Inhibition by Modified Oligodeoxynucleotides of the Expression of Type‐1 Plasminogen Activator Inhibitor in Human Endothelial Cells

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Abstract
Antisense phosphodiester and phosphorothioate oligodeoxynucleotides (23‐residue or 24‐residue oligodeoxynucleotides) were constructed for sequences of type‐1 plasminogen‐activator‐inhibitor mRNA to assess their capability to modulate type‐1 plasminogen‐activator‐inhibitor‐mediated fibrinolysis. Antisense oligodeoxynucleotides were targeted at the mRNA sequence coding a signal peptide, at a part of the reactive center Ile342–Pro349, and at an internally translated segment Asn265–Leu272. The effect of antisense oligonucleotides on the concentration of type‐1 plasminogen activator inhibitor in conditioned media and human endothelial cells was determined by the activity test with fibrin as a substrate, and by immunoprecipitation after metabolic labelling of cells with [35S]methionine. Three phosphorothioate oligodeoxynucleotides were specifically inhibitory while phosphodiester oligodeoxynucleotides with the same sequence did not show any activity. Phosphorothioate oligodeoxynucleotides 2, 4 and 6 inhibited the synthesis of type‐1 plasminogen activator inhibitor in endothelial cells in a time‐dependent and concentration‐dependent manner. These data suggest that antisense oligodeoxynucleotides may be useful in the design of antithrombolytic therapeutics.