IMMUNOCHEMICAL STRUCTURE FUNCTION OF APOLIPOPROTEIN A-I

载脂蛋白A-I的免疫化学结构功能

• 批准号: 6194795
• 负责人: Linda K Curtiss
• 金额: $ 44.33万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 1990
• 资助国家: 美国
• 起止时间: 1990-07-01 至 2004-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6194795
• 关键词: apolipoproteins; atherosclerosis; chemical kinetics; cholesterol; genetically modified animals; high density lipoproteins; human tissue; immunocytochemistry; laboratory mouse; macrophage; protein structure function

In atherosclerosis high density lipoproteins (HDL) are a key target for therapeutic intervention. This proposal will examine how apolipoprotein (apo) AI promotes efficient transfer of excess cholesterol (C) from peripheral cells to plasma HDL. Using mouse models of atherosclerosis, we will test the hypothesis that specificity for apoAI in mediating macrophage (Mphi) C efflux is a function of its ability to dissociate from spherical HDL. Dissociation gives rise to a stable, lipid-poor apoAI that can interact with cellular receptors to mediate energy-dependent C efflux. The first specific aim will examine the in vitro specificity of energy-dependent transport of C from cholesteryl ester loaded Mphi. Multiple exchangeable lipid-free and lipid-poor apoproteins including apoAI, apoAII, apoAIV, and apoE will be compared. The second aim will identify the form(s) of apoAI present in vivo in the intima of an atherosclerotic lesion using immunochemical and physicochemical analyses. The third aim will identify the in vivo mechanism(s) for formation of lipid-poor apoAI and will examine the roles of scavenger receptor-type B1, lipoprotein lipase, hepatic lipase, phospholipid transfer protein and apoAII. The fourth aim will test the hypothesis that the in vivo specificity requirement for apoAI can be bypassed by providing an alternate source of lipid-poor apoAI within lesions. Transgenic mice expressing Mphi-specific apoAI will be produced and will serve as donors in bone marrow reconstitution studies of atherosclerosis-prone mice. This hypothesis-driven proposal will: 1) provide a precise understanding of the lipoprotein composition of interstitial fluids; 2) identify in vivo mechanisms to explain the unique capacity of apoAI to remove C from lesions; and 3) use permanent gene transfer to alter the course of atherosclerotic vascular disease.

在动脉粥样硬化中，高密度脂蛋白（HDL）是治疗干预的关键靶标。 这项提案将研究载脂蛋白（apo）AI如何促进过量胆固醇（C）从外周细胞有效转移到血浆HDL。 使用小鼠动脉粥样硬化模型，我们将测试的假设，介导巨噬细胞（Mphi）C流出的apoAI的特异性是其从球形HDL解离的能力的函数。解离产生稳定的、脂质贫乏的apoAI，其可以与细胞受体相互作用以介导能量依赖性C流出。 第一个具体目标将检查C从胆固醇酯负载的Mphi的能量依赖性转运的体外特异性。 将比较多种可交换的无脂和贫脂载脂蛋白，包括apoAI、apoAII、apoAIV和apoE。 第二个目标是使用免疫化学和物理化学分析来鉴定动脉粥样硬化病变内膜中存在的apoAI的形式。 第三个目标是确定体内形成贫脂apoAI的机制，并检查清道夫受体B1型、脂蛋白脂酶、肝脂酶、磷脂转移蛋白和apoAII的作用。 第四个目标将测试这样的假设，即可以通过在病变内提供脂质贫乏的apoAI的替代来源来绕过apoAI的体内特异性要求。 将产生表达Mphi特异性apoAI的转基因小鼠，并将其作为动脉粥样硬化易感小鼠骨髓重建研究的供体。该假设驱动的提议将：1）提供对间质液的脂蛋白组成的精确理解; 2）确定体内机制以解释apoAI从病变中去除C的独特能力;和3）使用永久性基因转移来改变动脉粥样硬化血管疾病的病程。