APOA-I PRIMARY SEQUENCE IN HDL ASSEMBLY AND FUNCTION

HDL 组装和功能中的 APOA-I 主要序列

• 批准号: 2750524
• 负责人: TRUDY M FORTE
• 金额: $ 26.47万
• 依托单位: UNIVERSITY OF CALIF-LAWRENC BERKELEY LAB
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-08-01 至 2000-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2750524
• 关键词: animal genetic material tag; apolipoproteins; atherosclerosis; blood lipoprotein biosynthesis; cholesterol; disulfide bond; gene mutation; genetically modified animals; high density lipoproteins; human genetic material tag; laboratory mouse; macrophage; nutrition related tag; pathogenic diet; protein structure function

DESCRIPTION (Adapted from applicant's abstract): Several naturally occurring mutations in the lipid binding domain of apoAI are associated with a decreased HDL concentration but paradoxically are not associated with premature cardiovascular disease. One such mutation is the Cys for Arg substitution at amino acid 173, which occur in apoAI-M(ilano). The major aim of the present proposal is to create an animal model in which to study the anti-atherogenic potential of apoAI-M. A single copy, homologous recombination strategy will be employed to replace the mouse apoAI gene with the human apoAI or the apoAI-M gene to create mice having equivalent expression of human apoAI or apoAIM expression. The gene targeting procedure will be carried out using ES cells from strain C57BL/6, an atherosclerosis susceptible strain, in order to examine whether the apoAI-M replacement animals are protected from dietary induced atherosclerosis. The mice expressing the human apoAI will serve as controls in studying the mechanism of any protective effect of apoAI-M. The applicant will test the following hypotheses. (1) in the background of the atherosclerosis susceptible C57BL/6 mouse on an atherogenic diet, the apoAIM gene will be more protective against lesion development than the normal human apoAI. (2) the apoAI/apoAI-M genotype in "knock-in" mice will recapitulate the unusual lipid and lipoprotein profile found in humans, since apoAI-M had only been observed in heterozygous carriers. (3) apoAI-M HDL and/or a specific sub-population of variant HDL are significantly more efficient than normal apoAI in stimulating cholesterol efflux from mouse peritoneal macrophages, thus decreasing lipid accumulation in the artery wall. The second phase of the experiments are to elucidate whether apoAI-M dimerization on HDL particles produced by disulfide bond formation is linked with apoAIM's putative protective properties. An Arg-173 to Leu mutation will be introduced into mice in the same fashion as was used for apoAI-M. Other Arg-Cys substitutions in the lipid binding domains of apoAI will be produced to determine whether these substitutions, as a class, have the potential for being protective. These studies provide a novel approach for addressing the important human issues whether apoAI-M protects against atherosclerosis and whether other Arg-Cys substitutions have the same potential.

描述（改编自申请人的摘要）：自然而然 ApoAI的脂质结合结构域中发生突变与 HDL浓度降低，但自相矛盾地与 心血管疾病过早。 一个这样的突变是arg的cys 在Apoai-M（Ilano）中发生的氨基酸173的取代。 专业 本提案的目的是创建一个动物模型来研究 Apoai-M的抗动脉粥样硬化潜力。 一个副本，同源 将采用重组策略来代替小鼠apoai基因 人类apoai或apoai-m基因创建等效的小鼠 人类apoai或apoaim表达的表达。 基因靶向 将使用来自C57BL菌株/6菌株的ES细胞进行过程 动脉粥样硬化易感性菌株，以检查apoai-m是否是否 替代动物免受饮食诱导的动脉粥样硬化。 这 表达人apoai的小鼠将作为研究 Apoai-M的任何保护作用的机制。 申请人将检验以下假设。 （1）背景 动脉粥样硬化在动脉粥样硬化饮食中易感C57BL/6小鼠， Apoaim基因将对病变发育更具保护性 正常的人类apoai。 （2）“敲入”小鼠中的apoai/apoai-m基因型将 概括在人类中发现的异常脂质和脂蛋白谱， 由于仅在杂合载体中观察到Apoai-M。 （3）Apoai-M HDL和/或变体HDL的特定子群明显更多 从小鼠刺激胆固醇外排的高效 腹膜巨噬细胞，从而减少动脉中的脂质积累 墙。 实验的第二阶段是阐明apoai-m是否 二硫键形成产生的HDL颗粒上的二聚化是连接的 具有Apoaim的推定保护特性。 Arg-173到Leu突变 将以与Apoai-M相同的方式引入小鼠。 ApoAI的脂质结合域中的其他Arg-cys取代将是 生产以确定这些替代品是否具有 潜在的保护。 这些研究提供了一种新颖的方法 解决重要的人类问题是否保护 动脉粥样硬化以及其他ARG-CYS取代是否具有相同 潜在的。