Targeting the Foam Cell to Prevent Atherosclerosis

针对泡沫细胞预防动脉粥样硬化

基本信息

批准号：
8110006
负责人：
Antonio Paul
金额：
$ 39.5万
依托单位：
ALBANY MEDICAL COLLEGE
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-07-15 至 2015-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8110006
关键词：
Ablation Adenoviruses Adipose tissue Adverse effects Affect Antiatherogenic Apolipoprotein A-I Apolipoprotein E Apolipoproteins Apoptosis Arterial Fatty Streak Atherosclerosis Biology Cells Cellular biology Cholesterol Cholesterol Esters Co-Immunoprecipitations Data Deposition Development Diet Esters Exposure to Family Fatty acid glycerol esters Foam Cells Gene Expression Gene Silencing Genes Genomics Goals Human In Vitro Induction of Apoptosis Inflammation Inflammatory Response Knockout Mice Lead Lesion Lipid-Laden Macrophage Lipids Low-Density Lipoproteins Mediating Metabolism Methods Microarray Analysis Mitochondria Molecular Mouse Protein Mus Pharmaceutical Preparations Phospholipids Plasma Play Prevention Process Protein Deficiency Proteins Proteomics RNA RNA amplification Role Rupture Staging Stress Structure System Testing Therapeutic Therapeutic Intervention Thinking Toxic effect Work atherogenesis atheroprotective base cholesterol trafficking comparative extracellular feeding helper-dependent adenoviral vector hypercholesterolemia improved in vitro testing in vivo insight laser capture microdissection lipid transport loss of function macrophage member monolayer novel strategies prevent protein expression public health relevance research study response therapeutic target uptake

Ablation Adenoviruses Adipose tissue Adverse effects Affect Antiatherogenic Apolipoprotein A-I Apolipoprotein E Apolipoproteins Apoptosis Arterial Fatty Streak Atherosclerosis Biology Cells Cellular biology Cholesterol Cholesterol Esters Co-Immunoprecipitations Data Deposition Development Diet Esters Exposure to Family Fatty acid glycerol esters Foam Cells Gene Expression Gene Silencing Genes Genomics Goals Human In Vitro Induction of Apoptosis Inflammation Inflammatory Response Knockout Mice Lead Lesion Lipid-Laden Macrophage Lipids Low-Density Lipoproteins Mediating Metabolism Methods Microarray Analysis Mitochondria Molecular Mouse Protein Mus Pharmaceutical Preparations Phospholipids Plasma Play Prevention Process Protein Deficiency Proteins Proteomics RNA RNA amplification Role Rupture Staging Stress Structure System Testing Therapeutic Therapeutic Intervention Thinking Toxic effect Work atherogenesis atheroprotective base cholesterol trafficking comparative extracellular feeding helper-dependent adenoviral vector hypercholesterolemia improved in vitro testing in vivo insight laser capture microdissection lipid transport loss of function macrophage member monolayer novel strategies prevent protein expression public health relevance research study response therapeutic target uptake

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项目摘要

DESCRIPTION (provided by applicant): Summary Lipid-laden macrophages or "foam cells" play crucial roles in all the stages of atherogenesis, from lesion initiation to plaque rupture. However, our current therapeutic arsenal lacks of drugs primarily targeting the foam cell to prevent atherosclerosis development. Macrophages become foam cells by the uptake of modified low- density lipoproteins (mLDL) and accumulation of their lipids, mostly cholesterol (CHOL) esters (CE), in cytoplasmic lipid droplets (LDs). LDs consist of a core of neutral lipid stabilized and circumscribed by phospholipids, free CHOL (FC) and proteins. The main structural LD-associated proteins are the members of the PAT-family, and in macrophages the most abundant PAT-protein is adipose differentiation-related protein (ADFP). We have shown that ADFP plays a key role in foam cell formation, and its absence severely restricts the ability of macrophages to become foam cells in vitro and in vivo. ADFP is upregulated in human and mouse atherosclerotic lesions, and ADFP ablation in apolipoprotein E-null (apoE-/-) mice is atheroprotective. Enhancing CHOL efflux from macrophages, e.g. by increasing the concentration in plasma of CHOL acceptors or by upregulating the expression of CHOL efflux transporters, protects against atherosclerosis. This also seems to be the atheroprotective mechanism associated to the lack of ADFP, since ADFP-deficient macrophages do not accumulate as much CHOL in the form of CE in LDs, and efflux more CHOL to extracellular acceptors. The broad goal of this proposal is to gain insight on the role of ADFP in LD biology, foam cell formation and atherosclerosis development. In the first aim, we will test in vitro the response, in terms of inflammation and apoptosis, of ADFP-deficient macrophages cultured under high lipid stress conditions. We will also check if the atheroprotection observed in ADFP-/- mice fed regular chow is also observed when mice are challenged with a high fat/high CHOL diet. In the second aim, we will increase the plasma concentration of the CHOL acceptor apolipoprotein A1 (apoA1) with a helper-dependent adenovirus (HDAd) vector system, and assess whether two different approaches to increase CHOL efflux (i.e. increasing extracellular acceptors and limiting the cell's ability to store CHOL) will have additive atheroprotective effects. In the third aim, we will perform proteomic analyses to find the main LD-associated proteins and ADFP interactors in macrophages cultured under lipid-storing and lipid-efflux conditions. We will also perform a global gene expression analysis in foam cells directly isolated from lesions of mice that do or do not express ADFP to determine how the foam cell adjusts to the lack of ADFP. From this broad-based analysis of the role of ADFP in foam cell formation and atherosclerosis we expect to obtain a breadth of understanding that may help to set- up the bases for novel strategies to target the foam cell to prevent atherosclerosis development. PUBLIC HEALTH RELEVANCE: By elucidating the response of ADFP-deficient macrophages to high lipid stress conditions, whether preventing cholesterol accumulation in lipid droplets while increasing the ability of the extracellular milieu to accept cholesterol can work synergistically to prevent atherosclerosis, the molecular mechanisms that govern lipid droplet genesis and degradation in macrophages, how ADFP is involved in the processes, and how the foam cell adjust to the lack of ADFP, these studies will provide a breadth of understanding on foam cell biology that may be very useful if drugs to target the lipid droplet to prevent foam cell formation and atheroclerosis are to be developed.

描述（由申请人提供）：摘要载有脂质的巨噬细胞或“泡沫细胞”在动脉粥样硬化的所有阶段起着至关重要的作用，从病变启动到斑块破裂。但是，我们目前的治疗性阿森纳缺乏主要针对泡沫细胞的药物，以防止动脉粥样硬化的发展。巨噬细胞通过摄取改良的低密度脂蛋白（MLDL）和脂质的积累而变成泡沫细胞，在细胞质脂质液滴（LDS）中，其脂质（主要是胆固醇（CE））。 LD由中性脂质的核心组成，并由磷脂，游离CHOL（FC）和蛋白质限制。主要的结构LD相关蛋白是PAT家族的成员，在巨噬细胞中，最丰富的Pat蛋白是脂肪分化相关蛋白（ADFP）。我们已经表明，ADFP在泡沫细胞形成中起关键作用，并且它的缺失严重限制了巨噬细胞在体外和体内成为泡沫细胞的能力。 ADFP在人和小鼠动脉粥样硬化病变中被上调，载脂蛋白E-NULL（APOE-/ - ）小鼠中的ADFP消融是动脉粥样硬化的。增强巨噬细胞的chol外排，例如通过增加CHOL受体血浆的浓度或通过上调Chol外排转运蛋白的表达，可以预防动脉粥样硬化。这似乎也是与缺乏ADFP相关的动脉保护机制，因为ADFP缺陷型巨噬细胞在LDS中的CE形式不会积聚那么多的CHOL，而对细胞外受体则更多的CHOL。该提案的广泛目标是了解ADFP在LD生物学，泡沫细胞形成和动脉粥样硬化发展中的作用。在第一个目标中，我们将在体外测试在高脂质应激条件下培养的ADFP缺陷巨噬细胞的炎症和凋亡的反应。我们还将检查在高脂肪/高chol饮食挑战小鼠时，还观察到在ADFP - / - 小鼠中观察到的动脉保护作用。在第二个目标中，我们将增加具有辅助腺病毒（HDAD）矢量系统的CHOL受体载脂蛋白A1（APOA1）的血浆浓度，并评估两种不同的方法来增加chol外排（即增加细胞外受体的增加并限制细胞的能力，并限制了储存舌壳的能力）会添加效果。在第三个目标中，我们将进行蛋白质组学分析，以在脂质储存和脂质效能条件下培养的巨噬细胞中找到与LD相关的主要蛋白和ADFP相互作用者。我们还将在直接从表达或未表达ADFP的小鼠病变的泡沫细胞中进行全局基因表达分析，以确定泡沫细胞如何适应缺乏ADFP。从对ADFP在泡沫细胞形成和动脉粥样硬化中的作用的基于广泛的分析中，我们希望获得广泛的理解，这可能有助于建立新型策略的基础，以靶向泡沫细胞以防止动脉粥样硬化的发展。 PUBLIC HEALTH RELEVANCE: By elucidating the response of ADFP-deficient macrophages to high lipid stress conditions, whether preventing cholesterol accumulation in lipid droplets while increasing the ability of the extracellular milieu to accept cholesterol can work synergistically to prevent atherosclerosis, the molecular mechanisms that govern lipid droplet genesis and degradation in macrophages, how ADFP is involved in the过程，以及泡沫细胞如何适应缺乏ADFP，这些研究将为泡沫细胞生物学提供广泛的了解，如果要靶向脂质液滴以防止泡沫细胞的形成和动脉粥样硬化，则可能非常有用。