Adeno-associated virus mediated gene delivery into coronary microvessels of chronically instrumented dogs.
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.
Post
Heiner
H
Dept. of Physiology, Basic Sciences Bldg., Rm. 636, New York Medical College, Valhalla, NY 10595, USA.
Kajstura
Jan
J
Lei
Biao
B
Sessa
William C
WC
Byrne
Barry
B
Anversa
Piero
P
Hintze
Thomas H
TH
Recchia
Fabio A
FA
D000818:Animals; D003331:Coronary Vessels / Q000378:metabolism* / Q000502:physiology; D000229:Dependovirus; D004285:Dogs; D004730:Endothelium, Vascular / Q000166:cytology / Q000378:metabolism; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins; D006439:Hemodynamics; D007150:Immunohistochemistry; D007202:Indicators and Reagents / Q000493:pharmacokinetics; D007958:Leukocyte Count; D008164:Luminescent Proteins / Q000235:genetics / Q000493:pharmacokinetics; D018613:Microscopy, Confocal; D032389:Myocytes, Smooth Muscle / Q000378:metabolism; D009504:Neutrophils / Q000166:cytology; D014018:Tissue Distribution; D014161:Transduction, Genetic
D007202:Indicators and Reagents; D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins