title: In vivo visualization and attenuation of oxidized lipid accumulation in hypercholesterolemic zebrafish.
abstract: 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.
authors: Department of Medicine, UCSD, La Jolla, California 92093, USA.
authors: Longhou
authors: L
authors: Fang
authors: Simone R
authors: SR
authors: Green
authors: Ji Sun
authors: JS
authors: Baek
authors: Sang-Hak
authors: SH
authors: Lee
authors: Felix
authors: F
authors: Ellett
authors: Elena
authors: E
authors: Deer
authors: Graham J
authors: GJ
authors: Lieschke
authors: Joseph L
authors: JL
authors: Witztum
authors: Sotirios
authors: S
authors: Tsimikas
authors: Yury I
authors: YI
authors: Miller
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D000911:Antibodies, Monoclonal / Q000235:genetics / Q000276:immunology / Q000627:therapeutic use; D000924:Anticholesteremic Agents / Q000627:therapeutic use; D001808:Blood Vessels / Q000378:metabolism*; D002791:Cholesterol, Dietary / Q000493:pharmacokinetics*; D004195:Disease Models, Animal; D004353:Drug Evaluation, Preclinical; D000939:Epitopes / Q000276:immunology; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins / Q000032:analysis / Q000235:genetics; D006801:Humans; D006834:Hydrazines / Q000032:analysis; D006937:Hypercholesterolemia / Q000188:drug therapy / Q000378:metabolism* / Q000628:therapy; D007140:Immunoglobulin Fab Fragments / Q000235:genetics; D008077:Lipoproteins, LDL / Q000276:immunology / Q000378:metabolism*; D008264:Macrophages / Q000378:metabolism / Q000648:ultrastructure; D008315:Malondialdehyde / Q000031:analogs & derivatives* / Q000276:immunology / Q000378:metabolism; D018613:Microscopy, Confocal / Q000379:methods*; D008856:Microscopy, Fluorescence / Q000379:methods*; D010084:Oxidation-Reduction; D011162:Porphobilinogen / Q000031:analogs & derivatives / Q000032:analysis; D011341:Probucol / Q000627:therapeutic use; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology / Q000627:therapeutic use; D057127:Single-Chain Antibodies / Q000235:genetics / Q000276:immunology / Q000627:therapeutic use; D013696:Temperature; D014680:Veins / Q000378:metabolism; D015027:Zebrafish / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D029961:Zebrafish Proteins / Q000276:immunology / Q000378:metabolism*
chemical_list: C538930:3-anilino difluoroboron dipyrromethene; C417561:Alexa 488 hydrazide; D000911:Antibodies, Monoclonal; D000924:Anticholesteremic Agents; D002791:Cholesterol, Dietary; D000939:Epitopes; D006834:Hydrazines; D007140:Immunoglobulin Fab Fragments; D008077:Lipoproteins, LDL; D011993:Recombinant Fusion Proteins; D057127:Single-Chain Antibodies; D029961:Zebrafish Proteins; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D008315:Malondialdehyde; D011162:Porphobilinogen; D011341:Probucol