Macrophage Produced Phospholipid Transfer Protein (PLTP)

巨噬细胞产生磷脂转移蛋白 (PLTP)

基本信息

批准号：
8257889
负责人：
Linda K Curtiss
金额：
$ 23.69万
依托单位：
SCRIPPS RESEARCH INSTITUTE, THE
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-04-15 至 2014-01-01
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8257889
关键词：
ATP-Binding Cassette Transporters Acute-Phase Proteins Affect Agonist Anti-Inflammatory Agents Anti-inflammatory Antisense Oligonucleotides Aorta Arterial Fatty Streak Atherosclerosis Binding Bone Marrow Bone Marrow Cells Bone Marrow Transplantation Cell membrane Cells Cholesterol Chronic Chronic Disease Complement Coronary heart disease Diabetes Mellitus Diet Dimerization Disease Endosomes Energy Transfer Event Exhibits Fatty acid glycerol esters Gene Expression Genes Goals Heart Hepatic Hepatocyte High Density Lipoprotein Cholesterol Human In Vitro Inflammation Inflammatory Inflammatory Response Lasers Lead Lesion Leukocytes Lipids Lipoproteins Liver Mediating Membrane Membrane Microdomains Microscopy Monitor Mus Pathway interactions Phospholipid Transfer Proteins Phospholipids Physiological Physiology Plasma Play Poly I-C Production Regulation Reporting Research Risk Factors Role Scanning Signal Transduction Signal Transduction Pathway Sinus Sterols TLR2 gene TLR3 gene TNF gene Testing Therapeutic Tissues Yang atherogenesis atheroprotective autocrine base cardiovascular risk factor cytokine dimer feeding improved in vivo interstitial macrophage mouse model mutant novel overexpression paracrine plasma phospholipid transfer protein public health relevance reconstitution response sound

项目摘要

DESCRIPTION (provided by applicant): Macrophages (MF) play a central role in atherogenesis through accumulation of lipids and production of inflammatory cytokines. Plasma phospholipid transfer protein (PLTP) is associated with MF in lesions and its' expression is up-regulated by lipid loading. We reported that PLTP produced within lesions is atheroprotective. The long-term goal of this project is to identify mechanisms whereby cell associated PLTP is anti-inflammatory and atheroprotective. The first aim will test the hypothesis that MF PLTP disrupts TLR2/1 plasma membrane dimerization and signaling, but not preformed TLR3 dimer mediated signaling in endosomes. We will assess the anti-inflammatory effects of MF PLTP on TLR2/1 and TLR3 signaling in PLTP- /-LDLr-/- mice irradiated and reconstituted with bone-marrow (BM) from PLTP+/+ or PLTP-/- mice. Cytokine production and atherosclerosis will be studied. The second aim will test the hypothesis that PLTP alters cholesterol dependent membrane domains (lipid rafts). We will determine if wild-type (WT) or mutant PLTP (which binds ABCA1 but lacks lipid transfer activity) disrupts lipid rafts and/or inhibits TLR2/1 dimerization in vivo and in vitro. The third aim will test the idea that reduced hepatic production of PLTP to reduce plasma levels of PLTP will favor the beneficial effects of cell associated PLTP. We will assess the anti-inflammatory effects of PLTP on TLR2 signaling in LDLr-/- mice with suppressed, but not deleted, hepatic PLTP. This will be accomplished with in vivo administration of mouse PLTP antisense oligonucleotides (ASO). Novel, targeted mouse models for study of chronic inflammation are used to establish the physiological significance of our hypotheses and will be complemented by in vitro studies to identify the mechanisms involved in these physiological events. Laser scanning confocal immunofluorescent microscopy is used to characterize early atherosclerosis progression. Fluorescent resonance energy transfer (FRET) is used to characterize changes in lipid rafts in response to PLTP. Finally, the proposed studies will use antisense oligonucleotides (ASO) to favor the beneficial effects of anti-inflammatory cellular PLTP while reducing the undesirable affects of circulating plasma PLTP. It is imperative that we now know whether cell-directed or plasma-directed PLTP atheroprotective therapeutics should be pursued for human treatment of cardiovascular risk factors. PUBLIC HEALTH RELEVANCE: The proposed studies will significantly alter the direction of PLTP research by providing sound evidence of its role in the regulation of intracellular inflammatory pathways. The results of our studies will lead to a new and improved understanding of MF physiology and inflammation thus providing the bases for novel treatment options for atherosclerosis, diabetes and other chronic diseases that have a strong inflammatory component.

描述（由申请人提供）：巨噬细胞（MF）通过脂质的积累和炎症细胞因子的产生而在动脉粥样硬化中起着核心作用。血浆磷脂转移蛋白（PLTP）与病变中的MF相关，其表达被脂质负荷上调。我们报告说，病变内产生的PLTP是动脉保护性的。该项目的长期目标是确定与细胞相关的PLTP具有抗炎和动脉保护性的机制。第一个目的将检验以下假设：MF PLTP破坏TLR2/1质膜二聚和信号传导，但不会预先形成的TLR3二聚体介导的内体信号传导。我们将评估MF PLTP对PLTP-/-LDLR - / - 小鼠在PLTP-/-LDLR - / - 小鼠中对TLR2/1和TLR3信号传导的抗炎作用。将研究细胞因子的产生和动脉粥样硬化。第二个目标将检验PLTP改变胆固醇依赖性膜结构域（脂质筏）的假设。我们将确定野生型（WT）或突变体PLTP（结合ABCA1但缺乏脂质转移活性）是否会破坏脂质筏和/或抑制体内和体外的TLR2/1二聚体。第三个目标将测试减少PLTP肝产生以降低PLTP水平的想法，这将有利于相关PLTP的有益作用。我们将评估PLTP对LDLR - / - 小鼠TLR2信号传导的抗炎作用，但抑制但未被删除的肝PLTP。这将通过体内给药小鼠PLTP反义寡核苷酸（ASO）来实现。用于研究慢性炎症的新型小鼠模型用于确定我们的假设的生理意义，并通过体外研究来补充以识别这些生理事件所涉及的机制。激光扫描共聚焦免疫荧光显微镜用于表征早期的动脉粥样硬化进展。荧光共振能量转移（FRET）用于表征响应PLTP的脂质筏的变化。最后，拟议的研究将使用反义寡核苷酸（ASO）来利用抗炎细胞PLTP的有益作用，同时减少循环血浆PLTP的不可避免的影响。现在，我们必须知道是否应采用以细胞为导向或等离子体指导的PLTP动脉保护治疗剂来进行人体治疗心血管危险因素的治疗。公共卫生相关性：拟议的研究将通过提供其在调节细胞内炎症途径中的作用的可靠证据来显着改变PLTP研究的方向。我们的研究结果将导致对MF生理和炎症的新知识，从而为动脉粥样硬化，糖尿病和其他具有强烈炎症成分的慢性疾病提供新的治疗选择的基础。