Adeno-associated virus mediated gene delivery into coronary microvessels of chronically instrumented dogs

J Appl Physiol (1985). 2003 Oct;95(4):1688-94. doi: 10.1152/japplphysiol.00896.2002. Epub 2003 Jul 3.

Abstract

The objective of this study was to assess the potential of adeno-associated virus (AAV)-mediated gene delivery into coronary microvessels in vivo in a large animal. Ten mongrel dogs were chronically instrumented and allowed to recover for 10 days. Dogs were reanesthetized, and the aorta was constricted by a hydraulic occluder, whereby left ventricular (LV) pressure increased by 30% and left circumflex coronary artery blood flow by 50%. Recombinant AAV (serotype 2, CMV enhancer/chicken beta-actin promoter) encoding for green fluorescent protein (GFP) was injected as a bolus into the left atrium during aortic constriction at total titers of 1010 or 1012 infectious units. Dogs were followed for 2 (n = 4)or4wk(n = 6). Hemodynamics or body weight did not change. In LV tissue slices, a fluorescein-labeled antibody to GFP stained endothelial and smooth muscle cells but was absent in myocytes. To quantify transduction, slices were then stained with antibodies against alpha-smooth muscle actin or von Willebrand factor. Approximately 4% of arterioles and 2% of microvessels stained positive for anti-GFP independent from viral titer or duration. By regression analyses, the percent of vessels transfected was proportional to the increase in LV systolic pressure during occlusion. AAV is a potential vector for gene transfer into the coronary microcirculation in large animals, including perhaps humans.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Coronary Vessels / metabolism*
  • Coronary Vessels / physiology
  • Dependovirus*
  • Dogs
  • Endothelium, Vascular / cytology
  • Endothelium, Vascular / metabolism
  • Gene Transfer Techniques*
  • Genetic Vectors*
  • Green Fluorescent Proteins
  • Hemodynamics
  • Immunohistochemistry
  • Indicators and Reagents / pharmacokinetics
  • Leukocyte Count
  • Luminescent Proteins / genetics
  • Luminescent Proteins / pharmacokinetics
  • Microscopy, Confocal
  • Myocytes, Smooth Muscle / metabolism
  • Neutrophils / cytology
  • Tissue Distribution
  • Transduction, Genetic

Substances

  • Indicators and Reagents
  • Luminescent Proteins
  • Green Fluorescent Proteins