Journal Article

Stent-based delivery of AAV2 vectors encoding oxidation-resistant apoA1

Levy, Robert J., Zoltick, Philip W., Hooshdaran, Bahman, Fishbein, Ilia, Pressly, Benjamin B., Wilensky, Robert L., Gorman, Robert C., Alferiev, Ivan S., Smith, Jonathan D., Tschabrunn, Cory M.

2022

Overview

In-stent restenosis (ISR) complicates revascularization in the coronary and peripheral arteries. Apolipoprotein A1 (apoA1), the principal protein component of HDL possesses inherent anti-atherosclerotic and anti-restenotic properties. These beneficial traits are lost when wild type apoA1(WT) is subjected to oxidative modifications. We investigated whether local delivery of adeno-associated viral (AAV) vectors expressing oxidation-resistant apoA1(4WF) preserves apoA1 functionality. The efflux of 3 H-cholesterol from macrophages to the media conditioned by endogenously produced apoA1(4WF) was 2.1-fold higher than for apoA1(WT) conditioned media in the presence of hypochlorous acid emulating conditions of oxidative stress. The proliferation of apoA1(WT)- and apoA1(4FW)-transduced rat aortic smooth muscle cells (SMC) was inhibited by 66% ± 10% and 65% ± 11%, respectively, in comparison with non-transduced SMC (p < 0.001). Conversely, the proliferation of apoA1(4FW)-transduced, but not apoA1(WT)-transduced rat blood outgrowth endothelial cells (BOEC) was increased 41% ± 5% (p < 0.001). Both apoA1 transduction conditions similarly inhibited basal and TNFα-induced reactive oxygen species in rat aortic endothelial cells (RAEC) and resulted in the reduced rat monocyte attachment to the TNFα-activated endothelium. AAV2-eGFP vectors immobilized reversibly on stainless steel mesh surfaces through the protein G/anti-AAV2 antibody coupling, efficiently transduced cells in culture modeling stent-based delivery. In vivo studies in normal pigs, deploying AAV2 gene delivery stents (GDS) preloaded with AAV2-eGFP in the coronary arteries demonstrated transduction of the stented arteries. However, implantation of GDS formulated with AAV2-apoA1(4WF) failed to prevent in-stent restenosis in the atherosclerotic vasculature of hypercholesterolemic diabetic pigs. It is concluded that stent delivery of AAV2-4WF while feasible, is not effective for mitigation of restenosis in the presence of severe atherosclerotic disease.

• Publisher: Nature Publishing Group UK,Nature Publishing Group,Nature Portfolio
• Subject: 631/1647/2300 / 631/61/201 / 631/61/54 / 692/4019/2776 / 692/699/75/593/2728 / Animals / Aorta / Apolipoprotein A-I - genetics / Arteriosclerosis / Cell culture / Cholesterol / Coronary artery / Dependovirus - genetics / Diabetes mellitus / Endothelial Cells / Endothelium / Gene transfer / Genetic Vectors - genetics / High density lipoprotein / Humanities and Social Sciences / Hypochlorous acid / Implants / Macrophages / Monocytes / multidisciplinary / Oxidation / Oxidative stress / Protein G / Rats / Reactive oxygen species / Restenosis / Science / Science (multidisciplinary) / Smooth muscle / Stainless steel / Stents / Swine / Tumor necrosis factor-α / Veins & arteries