![]() ![]() Wild-type AAV infection does not produce any symptoms in humans ( 2). The use of adeno-associated viruses (AAV) has many advantages. Gene therapy with its potential for long-term correction of changes in protein expression in transduced cells represents a potential approach for treatment of vascular diseases ( 12), particularly if methods are developed to transduce microvessels, the site of vascular resistance and exchange, in large animals including humans. In the coronary circulation, microvascular endothelial dysfunction underlies and limits vascular function in hypertension ( 4), diabetes ( 16), and heart failure ( 4) and predicts a poor prognosis in patients with coronary heart disease ( 22). The endothelial cell layer plays a key role in maintaining vascular homeostasis by regulating vasomotion, leukocyte adherence, platelet activation, thrombosis, and vascular inflammation ( 24). AAV is a potential vector for gene transfer into the coronary microcirculation in large animals, including perhaps humans. By regression analyses, the percent of vessels transfected was proportional to the increase in LV systolic pressure during occlusion. Approximately 4% of arterioles and 2% of microvessels stained positive for anti-GFP independent from viral titer or duration. To quantify transduction, slices were then stained with antibodies against α-smooth muscle actin or von Willebrand factor. ![]() ![]() In LV tissue slices, a fluorescein-labeled antibody to GFP stained endothelial and smooth muscle cells but was absent in myocytes. Hemodynamics or body weight did not change. Dogs were followed for 2 ( n = 4)or4wk( n = 6). Recombinant AAV (serotype 2, CMV enhancer/chicken β-actin promoter) encoding for green fluorescent protein (GFP) was injected as a bolus into the left atrium during aortic constriction at total titers of 10 10 or 10 12 infectious units. 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%. Ten mongrel dogs were chronically instrumented and allowed to recover for 10 days. 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. ![]()
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