REACTIVE INTERMEDIATES OF OXIDATIVE LIPID FRAGMENTATION

氧化脂质断裂的反应中间体

基本信息

批准号：
9321185
负责人：
Robert Gerd Salomon
金额：
$ 38.31万
依托单位：
CASE WESTERN RESERVE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2006
资助国家：
美国
起止时间：
2006-05-01 至 2019-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9321185
关键词：
4 hydroxynonenal Acids Age related macular degeneration Alcohols Aldehydes Amination Amines Apoptosis Appearance Atrophic Basic Science Binding Biochemistry Biological Blood Blood Platelets Bone Marrow Brain Injuries Catalysis Cell Culture Techniques Cell membrane Cells Cellular Membrane Chemistry Choroidal Neovascularization Cytosol Diffuse Diffusion Docosahexaenoic Acids Endothelial Cells Equus caballus Ethanolamines Extracellular Matrix Generations Glean Glutathione Human In Vitro Individual Inflammatory Injury Intercept Interleukin-17 Investigation Lactones Leukotrienes Light Lipid Bilayers Lipids Measures Membrane Membrane Proteins Modeling Modification Neurons Oxides Pathogenesis Pathway interactions Patients Phospholipids Photoreceptors Physiological Pilot Projects Post-Translational Protein Processing Proteins Rattus Reaction Research Retina Retinal Scheme Signal Pathway Signal Transduction Site T-Lymphocyte TLR1 gene TLR2 gene TP53 gene Testing Therapeutic immunosuppression Tissues Tube VEGFA gene adduct angiogenesis clinically significant deacylation design extracellular geographic atrophy hydrophilicity in vivo insight macrophage monolayer novel therapeutics oxidation oxidative damage oxidized lipid photoreceptor disc prevent public health relevance receptor remote location retinal damage retinal rods

项目摘要

DESCRIPTION (provided by applicant): Docosahexaenoate phospholipids (DHA-PLs) are uniquely abundant in retinal and neuronal cells. Our basic research led to the discoveries that oxidation of DHA-PLs results in the generation of carboxyethylpyrrole (CEP) modifications of proteins and ethanolamine phospholipids whose levels are elevated in retinas and blood from individuals with age-related macular degeneration (AMD). Subsequent research revealed that CEPs stimulate angiogenesis found in the choroidal neovascularization of "wet" AMD through a vascular endothelial growth factor-independent mechanism involving Toll-like receptor (TLR)2 signaling. CEPs also contribute to the global retinal atrophy of "dry" AMD by IFNᵞ and IL-17-producing CEP-specific T cells that promote M1 polarization of macrophages in the retina. Our most recent studies revealed that oxidized DHA-PLs release HOHA-lactone that can dissociate from cell membranes and react with proteins to generate CEPs, previously only known to be produced by direct reaction of an oxidized DHA-PLs with proteins. We now propose studies of HOHA-lactone chemistry and transport through cell membranes and monolayers to evaluate the likelihood that its escape from DHA-rich membranes of photoreceptor rod cell disks can produce CEPs in locations remote from the site of membrane oxidation. This may contribute to the clinically significant elevated levels of CEPs we discovered in the blood of individuals with AMD and it may account for CEP generation in the blood of rats upon light-induced oxidative injury of their retinas. We will examine the possibility that HOHA-lactone can enter cells and generate CEP modifications of intracellular proteins that can bind with and activate intracellular receptors such as platelet TLR9. Studies of HOHA-lactone glutathione (GSH) Michael adduct biochemistry will test the hypotheses that this adduct can serve as a Trojan horse that transports a CEP precursor out of cells, and that in conjunction with ALD-catalyzed reduction, can prevent CEP formation. Inspired by the biological activities found previously for adducts of other oxidized lipids with GSH, e.g. leukotrienes, pilot studies were conducted with GSH adducts of HOHA-lactone that revealed that submicromolar concentrations of GSH-HOHA-lactone and the alcohol produced by reduction of this aldehyde stimulate proliferation and tube formation by HUVEC cells. New cell biological studies are proposed to investigate the effects of HOHA-lactone, its GSH adducts and the CEP modifications of proteins and ethanol-amine phospholipids derived from the HOHA-lactone on primary human RPE cells, bone marrow-derived macrophages, and primary choroidal endothelial cells, including studies on the signaling pathways leading to the biological effects. The potential utility of the mechanistic information to be gleaned from the in vitro and in vivo studies proposed is exemplified by new insights recently developed suggesting that immunosuppressive therapy might be effective for ameliorating the retinal damage of "dry" AMD caused by a CEP-induced T-cell promoted invasion of the retina by inflammatory macrophages and their CEP-potentiated activation.

描述（由应用程序提供）：视网膜和神经元细胞中二十六烯酸酯磷脂（DHA-PLS）具有独特的丰富。我们的基础研究导致发现DHA-PLS的氧化导致蛋白质和乙醇胺磷脂的修饰产生，它们的水平在视网膜中升高，并从患有年龄相关的黄斑变性（AMD）的个体中升高。随后的研究表明，CEPS通过血管内皮生长因子非依赖性机制（TLR）2信号传导的血管内皮生长因子非依赖性机制刺激“湿” AMD的脉络膜新生血管生成。 CEP还有助于IFNᵞ和IL-17产生的CEP特异性T细胞的“干” AMD的全局视网膜萎缩，从而促进视网膜中巨噬细胞的M1极化。我们的最新研究表明，氧化的DHA-PLS释放了Hoha-lactone，可以从细胞膜中解离并与蛋白质反应以产生CEP，以前仅通过氧化物DHA-PLS与蛋白质直接反应而产生。现在，我们提出了Hoha-lactone化学和通过细胞膜和单层传输的研究，以评估其从富含DHA的光感受器杆细胞盘逃脱的可能性，可以在远离膜氧化位置的位置产生CEP。这可能有助于我们在AMD患者血液中发现的CEP的临床显着升高水平，并且在光诱导的视网膜氧化性损伤后，大鼠血液中的CEP产生。我们将研究Hoha-lactone可以进入细胞并生成细胞内蛋白的CEP修饰的可能性，这些蛋白可以与细胞内受体结合并激活细胞内受体例如血小板TLR9。 Hoha-lactone谷胱甘肽（GSH）Michael Adruct生物化学的研究将测试该合并可以用作将CEP前体从细胞中运送出来的特洛伊木马，并且与ALD催化的还原相结合可以防止CEP形成。受到以前针对其他氧化脂质加合物的生物学活性的启发，例如白第三种，使用Hoha-lactone的GSH加合物进行了试点研究，该加合物揭示了粒度极浓度的GSH-HOHA-LACTONE浓度以及通过减少这种醛来产生的酒精刺激了HUVEC细胞的增殖和管形成。提出了新的细胞生物学研究，以研究Hoha-lactone，其GSH加合物以及蛋白质的CEP修饰以及源自Hoha-lactone对原代人RPE细胞的乙醇 - 胺磷脂的修饰，骨髓骨髓衍生的巨噬细胞以及主要的脉络膜内皮内皮细胞，包括在信号path上的研究。机械信息的潜在效用最近开发的新见解表明，从体外和体内研究中收集的研究表明，免疫抑制疗法可能有效地改善由CEP诱导的T-Cell引起的“干燥” AMD的视网膜损害，从而促进了炎症性巨噬细胞及其CEP型促进视网膜的入侵，并促进了其CEP型巨噬细胞及其CEP诱导的侵袭。