New Avenues in GPIHBP1 Research

项目总结/摘要 本提案的目的是确定GPIHBP 1是否对脂肪分解特别重要。 处理载脂蛋白B48的脂蛋白，了解Gpihbp 1缺陷的动脉粥样硬化 （Gpihbp 1-/-）小鼠，并建立和分析天然存在的GPIHBP 1基因敲入小鼠模型 人类的突变 我们最近发现Gpihbp 1-/-小鼠有乳糜微粒血症，即使在普通饲料中，血浆甘油三酯 水平高达8000 mg/dl，血浆胆固醇水平高达800 mg/dl。因为GPIHBP 1是 位于“脂解组织”的毛细血管的管腔表面（例如，心脏，肌肉，脂肪），因为 GPIHBP 1结合脂蛋白脂酶（LPL）和乳糜微粒，我们怀疑GPIHBP 1作为一种蛋白酶， 用于代谢处理富含磷脂酰肌醇的脂蛋白的平台。值得注意的是，GPIHBP 1包含强烈的 带负电荷的氨基末端结构域，其对于结合LPL和脂蛋白两者都很重要。 GPihbp 1-/-小鼠血浆中含有载脂蛋白B48的脂蛋白水平升高，但正常水平 含有载脂蛋白B100的脂蛋白。我推测GPIHBP 1可能在这一过程中起着特别重要的作用。 加工含有载脂蛋白B48的脂蛋白。为了探索这一假设，我将检查 Apob中“apo-B100-only”或“apo-B48-only”突变纯合的GPihbp 1-/-小鼠。 大的脂蛋白，如乳糜微粒，通常被认为是非致动脉粥样硬化的，但我有 结果表明，喂食饲料的Gpihbp 1-/-小鼠发生自发性动脉粥样硬化病变。以确定是否 Gpihbp 1-/-小鼠的动脉粥样硬化是由“富含胆固醇”脂蛋白的胆固醇含量驱动的， 将检查依折麦布和Npc 1 l1缺乏对Gpihbp 1-/-小鼠对以下疾病易感性的影响： 动脉粥样硬化 最近，GPIHBP 1的G56 R和Q115 P纯合突变在患有 乳糜微粒血症。这两种突变都发生在高度保守的残基上。我发现Q115 P突变 GPIHBP 1与LPL或乳糜微粒结合的能力消失，但G56 R突变没有检测到 效果-通过体外试验判断。这些发现让我不确定是否在体外 GPIHBP 1的测定是GPIHBP 1在体内功能的真正可靠的指标。为了探讨这个问题，我将 生成并表征G56 R和Q115 P突变的敲入小鼠模型。 该建议的三个具体目标是：（1）确定GPIHBP 1是否对 清除含载脂蛋白B48的脂蛋白;（2）确定在喂食普通饲料的Gpihbp 1- /-小鼠由“富含胆固醇的”脂蛋白的胆固醇含量驱动;和（3）产生敲入小鼠 GPIHBP 1中G56 R和Q115 P突变的模型。与公共卫生的关系 了解脂蛋白代谢是重要的，因为血浆脂蛋白提供甘油三酯燃料， 因为脂蛋白是动脉粥样硬化形成的原因。该项目的一个主要目标是 了解GPIHBP 1在脂蛋白脂肪酶介导的富含磷脂酰肌醇的脂蛋白加工中的作用。 拟议的研究可能会提供有关脂质营养物质如何传递到重要组织的新信息; 我的实验还将探索Gpihbp 1缺乏时动脉粥样硬化的机制， 定义特定人GPIHBP 1错义突变的体内相关性。

PROJECT SUMMARY/ABSTRACT The objectives of this proposal are to determine if GPIHBP1 is particularly important for the lipolytic processing of apo-B48-containing lipoproteins, to understand the atherosclerosis in Gpihbp1-deficient (Gpihbp1-/-) mice, and to create and analyze knock-in mouse models for naturally occurring GPIHBP1 mutations in humans. We recently found that Gpihbp1-/- mice have chylomicronemia, even on chow diet, with plasma triglyceride levels as high as 8000 mg/dl and plasma cholesterol levels as high as 800 mg/dl. Because GPIHBP1 is located on the luminal surface of capillaries of "lipolytic tissues" (e.g., heart, muscle, fat) and because GPIHBP1 binds both lipoprotein lipase (LPL) and chylomicrons, we suspect that GPIHBP1 serves as a platform for the metabolic processing of triglyceride-rich lipoproteins. Of note, GPIHBP1 contains a strongly negatively charged amino-terminal domain that is important for binding both LPL and lipoproteins. Gpihbp1-/- mice have elevated levels of apo-B48-containing lipoproteins in their plasma, but normal levels of apo-B100-containing lipoproteins. I hypothesize that GPIHBP1 may have a particularly important role in the processing of apo-B48-containing liporoteins. To explore this hypothesis, I will examine the phenotypes of Gpihbp1-/- mice that are homozygous for the "apo-B100-only" or "apo-B48-only" mutations in Apob. Large lipoproteins such as chylomicrons are generally assumed to be nonatherogenic, but I have demonstrated that chow-fed Gpihbp1-/- mice develop spontaneous atheroscerotic lesions. To determine if the atherosclerosis in Gpihbp1-/- mice is driven by the cholesterol content of the "triglyceride-rich" lipoproteins, I will examine the impact of ezetimibe and Npc1l1 deficiency on the susceptibility of Gpihbp1-/- mice to atherosclerosis. Recently, homozygous G56R and Q115P mutations in GPIHBP1 were identified in patients with chylomicronemia. Both mutations occurred in highly conserved residues. I found that the Q115P mutation abolishes the ability of GPIHBP1 to bind to LPL or chylomicrons, but the G56R mutation had no detectable effect-as judged by in vitro assays. These findings left me with uncertainty regarding whether the in vitro assays of GPIHBP1 were truly reliable indicators of GPIHBP1 function in vivo. To explore this issue, I will generate and characterize knock-in mouse models for the G56R and the Q115P mutations. The three specific aims of this proposal are: (1) to determine if GPIHBP1 is particularly important for the clearance of apo-B48-containing lipoproteins; (2) to determine if atherosclerotic lesions in chow-fed Gpihbp1- /- mice are driven by the cholesterol content of "triglyceride-rich" lipoproteins; and (3) to create knock-in mouse models for the G56R and Q115P mutations in GPIHBP1. RELEVANCE TO PUBLIC HEALTH Understanding lipoprotein metabolism is important because plasma lipoproteins deliver triglyceride fuel to vital tissues and because lipoproteins are causal factors in atherogenesis. A key goal of this project is to understand the role of GPIHBP1 in the lipoprotein lipase-mediated processing of triglyceride-rich lipoproteins. The proposed studies are likely to provide new information on how lipid nutrients are delivered to vital tissues; my experiments will also explore mechanisms for atherosclerosis in the setting of Gpihbp1 deficiency and define the in vivo relevance of specific human GPIHBP1 missense mutations.