Cardiac-targeting transfection of tissue-type plasminogen activator gene to prevent the graft thrombosis and vascular anastomotic restenosis after coronary bypass

Thromb Res. 2014 Aug;134(2):440-8. doi: 10.1016/j.thromres.2014.04.018. Epub 2014 Apr 29.

Abstract

Aim: To observe the tissue-type plasminogen activator gene (t-PA) plasmid packaged with albumin nanoparticles crosslinked to albumin ultrasound microbubbles for targeting transfection to myocardium to prevent the graft thrombosis and vascular anastomotic restenosis after coronary bypass.

Methods: A dog model of coronary bypass using the autoallergic saphenous vein as the graft was made. A highly expressive t-PA gene plasmid packaged with albumin nanoparticles crosslinked to albumin ultrasound microbubbles was constructed. Targeting myocardial transfection was performed with this gene vector under the aid of therapeutic ultrasound(1MHz, 1.5 w/cm2, 6minutes, intravenously) after the bypass. The expression of t-PA in myocardium was detected with a multiclonal antibody to t-PA by the indirect immunohistochemical method. Venous blood t-PA and D-dimer contents were tested before and 1, 2 and 4weeks after the operation. The effects of this gene vector on thrombosis of the grafts and the coronary intimal hyperplasia around the anastomotic stoma were observed using a routine pathological examination, a morphometry for intimal thickness and area and the immuno-histochemical stain with a monoclonal antibody to PCNA for estimating the intimal SMC proliferation.

Results: The effective expression of t-PA protein by myocardium was obtained, followed by the persistent raises of blood t-PA and D-dimer 1, 2 and 4weeks after the transfection. Thrombosis of the grafts was successfully restrained. The expression of PCNA by coronary intimal vSMCs and intimal hyperplasia were remarkablely reduced.

Conclusion: This t-PA gene targeting vector could be used to prevent the dog thrombosis, which provided the experimental identification for prevention on human thrombotic diseases.

Keywords: Coronary bypass; Tissue-type plasminogen activator; albumin nanoparticles; thrombosis; ultrasound microbubbles.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Coronary Artery Bypass / adverse effects*
  • Dogs
  • Genetic Therapy / methods
  • Graft Occlusion, Vascular / etiology
  • Graft Occlusion, Vascular / genetics
  • Graft Occlusion, Vascular / prevention & control*
  • Humans
  • Male
  • Microbubbles
  • Nanoparticles / chemistry
  • Plasmids / administration & dosage
  • Plasmids / genetics
  • Plasmids / therapeutic use*
  • Thrombosis / etiology
  • Thrombosis / genetics
  • Thrombosis / prevention & control*
  • Tissue Plasminogen Activator / genetics*
  • Transfection* / methods
  • Ultrasonics

Substances

  • Tissue Plasminogen Activator