In vivo visualization and attenuation of oxidized lipid accumulation in hypercholesterolemic zebrafish

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.
J. Clin. Invest.   121(12): 4861-9 (Journal)
Registered Authors
Ellett, Felix, Lieschke, Graham J., Miller, Yury
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*
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.

Genes / Markers
Show all Figures
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Engineered Foreign Genes