In vivo visualization and attenuation of oxidized lipid accumulation in hypercholesterolemic zebrafish
- Authors
- Fang, L., Green, S.R., Baek, J.S., Lee, S.H., Ellett, F., Deer, E., Lieschke, G.J., Witztum, J.L., Tsimikas, S., and Miller, Y.I.
- ID
- ZDB-PUB-111129-15
- Date
- 2011
- Source
- J. Clin. Invest. 121(12): 4861-9 (Journal)
- Registered Authors
- Ellett, Felix, Lieschke, Graham J., Miller, Yury
- Keywords
- none
- MeSH Terms
-
- Animals
- Animals, Genetically Modified
- Antibodies, Monoclonal/genetics
- Antibodies, Monoclonal/immunology
- Antibodies, Monoclonal/therapeutic use
- Anticholesteremic Agents/therapeutic use
- Blood Vessels/metabolism*
- Cholesterol, Dietary/pharmacokinetics*
- Disease Models, Animal*
- Drug Evaluation, Preclinical
- Epitopes/immunology
- Genes, Reporter
- Green Fluorescent Proteins/analysis
- Green Fluorescent Proteins/genetics
- Humans
- Hydrazines/analysis
- Hypercholesterolemia/drug therapy
- Hypercholesterolemia/metabolism*
- Hypercholesterolemia/therapy
- Immunoglobulin Fab Fragments/genetics
- Lipoproteins, LDL/immunology
- Lipoproteins, LDL/metabolism*
- Macrophages/metabolism
- Macrophages/ultrastructure
- Malondialdehyde/analogs & derivatives*
- Malondialdehyde/immunology
- Malondialdehyde/metabolism
- Microscopy, Confocal/methods*
- Microscopy, Fluorescence/methods*
- Oxidation-Reduction
- Porphobilinogen/analogs & derivatives
- Porphobilinogen/analysis
- Probucol/therapeutic use
- Recombinant Fusion Proteins/genetics
- Recombinant Fusion Proteins/immunology
- Recombinant Fusion Proteins/therapeutic use
- Single-Chain Antibodies/genetics
- Single-Chain Antibodies/immunology
- Single-Chain Antibodies/therapeutic use
- Temperature
- Veins/metabolism
- Zebrafish/genetics
- Zebrafish/growth & development
- Zebrafish/metabolism*
- Zebrafish Proteins/immunology
- Zebrafish Proteins/metabolism*
- PubMed
- 22105168 Full text @ J. Clin. Invest.
Oxidative modification of LDL is an early pathological event in the development of atherosclerosis. Oxidation events such as malondialdehyde (MDA) formation may produce specific, immunogenic epitopes. Indeed, antibodies to MDA-derived epitopes are widely used in atherosclerosis research and have been demonstrated to enable cardiovascular imaging. In this study, we engineered a transgenic zebrafish with temperature-inducible expression of an EGFP-labeled single-chain human monoclonal antibody, IK17, which binds to MDA-LDL, and used optically transparent zebrafish larvae for imaging studies. Feeding a high-cholesterol diet (HCD) supplemented with a red fluorescent lipid marker to the transgenic zebrafish resulted in vascular lipid accumulation, quantified in live animals using confocal microscopy. After heat shock–induced expression of IK17-EGFP, we measured the time course of vascular accumulation of IK17-specific MDA epitopes. Treatment with either an antioxidant or a regression diet resulted in reduced IK17 binding to vascular lesions. Interestingly, homogenates of IK17-EGFP–expressing larvae bound to MDA-LDL and inhibited MDA-LDL binding to macrophages. Moreover, sustained expression of IK17-EGFP effectively prevented HCD-induced lipid accumulation in the vascular wall, suggesting that the antibody itself may have therapeutic effects. Thus, we conclude that HCD-fed zebrafish larvae with conditional expression of EGFP-labeled oxidation-specific antibodies afford an efficient method of testing dietary and/or other therapeutic antioxidant strategies that may ultimately be applied to humans.