Adeno-associated virus-mediated gene transfer of a secreted decoy human macrophage scavenger receptor reduces atherosclerotic lesion formation in LDL receptor knockout mice

Abstract

Macrophage scavenger receptors (MSR) promote atherosclerotic lesion formation, and modulation of MSR activity has been shown to influence atherosclerosis. Soluble receptors are effective in inhibiting receptor-mediated functions in various diseases. We have generated a secreted macrophage scavenger receptor (sMSR) that consists of the bovine growth hormone signal sequence and the human MSR A I extracellular domains. sMSR reduces degradation of atherogenic modified low-density lipoproteins and monocyte/macrophage adhesion on endothelial cells in vitro. To test long-term effects of sMSR, atherosclerosis-susceptible LDLR knockout mice were transduced via the tail vein with an adeno-associated virus (AAV) expressing sMSR or control enhanced green fluorescent protein (EGFP), and a Western-type diet was started. Gene transfer caused a temporary elevation in alkaline phosphatase and aspartate amino transferase values without a change in C-reactive protein. sMSR protein was detected in the plasma of the transduced mice by a specific ELISA 6 months after the gene transfer. AAV-mediated sMSR gene transfer reduced atherosclerotic lesion area in the aorta by 21% (P < 0.05) compared to EGFP-transduced control mice. Even though eradication of established disease was not possible, atherosclerotic lesion formation could be modified using AAV-mediated gene transfer of the decoy sMSR.

Nucleolin Is a Receptor for Maleylated-Bovine Serum Albumin on Macrophages

“…Jalkanen et al reported that soluble SRs inhibit receptor-mediated functions in atherosclerosis. 28) Those authors generated a secreted macrophage scavenger receptor (sMSR) that consisted of the bovine growth hormone signal sequence and the human MSR-AI extracellular domains. sMSRs reduce the degradation of atherogenic modified LDL and monocyte/macrophage adhesion on endothelial cells.…”

“…Even though eradication of established disease was not possible, atherosclerotic lesion formation could be modified using AAVmediated gene transfer of the sMSR. 28) As nucleolin is also detected in serum, 29) nucleolin operation in vivo might also be a useful means of preventing and treating SRs-involved diseases, including atherosclerosis.…”

Induced Pluripotent Stem Cell Clones Reprogrammed via Recombinant Adeno-Associated Virus-Mediated Transduction Contain Integrated Vector Sequences

“…rAAVs were produced as described by Zolotukhin et al (36). Briefly, for each virus, 23 ϫ 10 6 293T cells in total were plated into three 15-cm dishes and transfected with gene-carrying plasmids, constructed on the basis of the aforementioned pSubCMV-WPRE and pSub-CAG-WPRE plasmids, and with a serotype-determining helper plasmid(s) (pDG [for rAAV2] [10] or the combination of pBS-E2A-VA-E4 and p5e18-VD2/9 [for rAAV9] [8,15]), using JetPEI (Polyplus Transfection) according to the manufacturer's instructions. Cells were collected 2 days later and resuspended in lysis buffer (150 mM NaCl, 50 mM Tris, pH 8.5).…”

Antioxidant Gene Therapy for Cardiovascular Disease

“…57 In addition, adeno-associated virus-based gene transfer of soluble macrophage scavenger receptor AI reduced aortic lesion area. 58 However, the effects on atherosclerotic lesions were relatively modest, which is likely to be caused by the limitations in gene transfer efficiency and the mild phenotype of LDL receptor-deficient mice. In addition, redundancy with other scavenger receptors, particularly CD36, and the fact that macrophage scavenger receptor AI does not recognize all forms of modified LDL limit the efficacy.…”