Zebrafish models of vascular inflammation and atherosclerosis

血管炎症和动脉粥样硬化的斑马鱼模型

基本信息

批准号：
8320140
负责人：
Yury Miller
金额：
$ 41.95万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN DIEGO
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-09-01 至 2015-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8320140
关键词：
Animal Model Animals Antibodies Antioxidants Apolipoproteins B Arachidonate 15-Lipoxygenase Arterial Fatty Streak Arts Atherosclerosis Binding Blood Vessels Cholesterol Cholesterol Esters Collaborations Deposition Detection Development Diet Disease Endothelial Cells Epitopes Fab Immunoglobulins Foam Cells Goals Grant Heat-Shock Response Human Hyperglycemia Image Imaging Techniques Inflammation Inflammatory Inflammatory Response Injection of therapeutic agent Label Larva Lesion Life Lipid-Laden Macrophage Lipids Lipoproteins Lipoxygenase Inhibitors Low Density Lipoprotein oxidation Low-Density Lipoproteins Malondialdehyde Mass Spectrum Analysis Mediating Microscope Modeling Monitor Monoclonal Antibodies Myeloid Cells Myocardial Infarction Nonesterified Fatty Acids Optics Pathologic Pathway interactions Phospholipids Process Proteins Research Role Rupture Screening procedure Staging Stroke Techniques Testing Therapeutic Therapeutic Effect Time Transgenic Organisms Zebrafish atherogenesis cardiovascular imaging drug candidate feeding in vivo inhibitor/antagonist macrophage novel novel therapeutics oxidation oxidized lipid oxidized low density lipoprotein prevent promoter research study tool vascular inflammation

项目摘要

DESCRIPTION (provided by applicant): Animal experimental and human atherosclerosis studies have convincingly demonstrated that low- density lipoprotein (LDL) undergoes oxidation, which greatly enhances its atherogenicity. One of the major pathways of LDL oxidation in vivo is the oxidation catalyzed by 12/15-lipoxygenase (12/15LO). Because oxidized LDL (OxLDL) induces many inflammatory responses in the vascular wall, high levels of so-called oxidation-specific epitopes in atherosclerotic lesions often indicate that these lesions are prone to rupture, inducing myocardial infarction or stroke. Identification of vulnerable atherosclerotic plaques prone to rupture is a major challenge for cardiovascular imaging, as current imaging techniques provide little information on plaque composition. Our group is currently developing new imaging approaches using antibodies that specifically bind oxidation-specific epitopes. In addition to imaging applications, oxidation-specific antibodies are emerging as a therapeutic treatment of atherosclerosis because they prevent OxLDL's inflammatory effects. In this application, we propose to develop new zebrafish (Danio rerio) models to study mechanisms and pathologic effects of lipoprotein oxidation, new imaging techniques targeting oxidation-specific epitopes in the vasculature of live animals, as well as novel therapeutic strategies to diminish lipoprotein oxidation and its pathologic effects. Specifically, we propose: (1) To develop imaging and analytic techniques for in vivo and ex vivo study of lipoprotein oxidation and detection of oxidation-specific epitopes in vascular lesions. We will target three common classes of oxidation-specific epitopes: malondialdehide, oxidized phospholipids and oxidized cholesteryl esters. In one approach, we will inject i.v. zebrafish larvae with fluorescently labeled antibodies and use a confocal microscope to detect antibodies binding in vascular lesions. The second approach will be to generate transgenic zebrafish with conditional expression of GFP-labeled oxidation-specific antibodies. We will also use mass spectrometry techniques to identify oxidized lipids in apoB and apoA1 lipoproteins as well as those bound to oxidation-specific antibodies. (2) To create a model of 12/15LO-induced lipoprotein oxidation and to study in vivo its role in vascular lipid accumulation and inflammation. Vascular lesions in transgenic zebrafish with endothelial or myeloid cell specific expression of human 12/15LO will be analyzed for accumulation of oxidized lipids, macrophage recruitment and foam cell formation. (3) To test the therapeutic potential of oxidation-specific antibodies, 12/15LO inhibitors and antioxidants. In summary, in order to contribute to "enhancing zebrafish research with research tools and techniques", we propose to use the zebrafish model, including new transgenic lines as well as state-of-art imaging and mass spectrometry techniques, to study an important atherogenic process - lipoprotein oxidation.

描述（申请人提供）：动物实验和人体动脉粥样硬化研究令人信服地证明，低密度脂蛋白（LDL）会发生氧化，从而大大增强其致动脉粥样硬化性。体内LDL氧化的主要途径之一是12/15-脂氧合酶(12/15LO)催化的氧化。由于氧化低密度脂蛋白（OxLDL）会在血管壁中诱发多种炎症反应，因此动脉粥样硬化病变中所谓氧化特异性表位的高水平往往表明这些病变易于破裂，诱发心肌梗塞或中风。识别容易破裂的易损动脉粥样硬化斑块是心血管成像的主要挑战，因为当前的成像技术提供的有关斑块成分的信息很少。我们的小组目前正在开发新的成像方法，使用特异性结合氧化特异性表位的抗体。除了成像应用之外，氧化特异性抗体也正在作为动脉粥样硬化的治疗方法出现，因为它们可以防止 OxLDL 的炎症效应。在此应用中，我们建议开发新的斑马鱼（Danio rerio）模型来研究脂蛋白氧化的机制和病理效应，针对活体动物脉管系统中氧化特异性表位的新成像技术，以及减少脂蛋白氧化的新治疗策略及其病理作用。具体来说，我们建议：（1）开发用于脂蛋白氧化的体内和离体研究以及血管病变中氧化特异性表位检测的成像和分析技术。我们将针对三类常见的氧化特异性表位：丙二醛、氧化磷脂和氧化胆固醇酯。在一种方法中，我们将静脉注射。斑马鱼幼虫带有荧光标记的抗体，并使用共聚焦显微镜检测血管病变中的抗体结合。第二种方法是产生条件表达 GFP 标记的氧化特异性抗体的转基因斑马鱼。我们还将使用质谱技术来鉴定 apoB 和 apoA1 脂蛋白中的氧化脂质以及与氧化特异性抗体结合的脂质。 (2)建立12/15LO诱导的脂蛋白氧化模型并研究其在体内血管脂质积累和炎症中的作用。将分析具有人 12/15LO 的内皮细胞或骨髓细胞特异性表达的转基因斑马鱼的血管损伤，以了解氧化脂质的积累、巨噬细胞的募集和泡沫细胞的形成。 (3)测试氧化特异性抗体、12/15LO抑制剂和抗氧化剂的治疗潜力。总之，为了“利用研究工具和技术加强斑马鱼研究”，我们建议使用斑马鱼模型，包括新的转基因品系以及最先进的成像和质谱技术，来研究一种重要的致动脉粥样硬化斑马鱼的研究。过程——脂蛋白氧化。