Glutathionylated Products of Radical-Induced Lipid Oxidation in Inflammatory Disease

炎症性疾病中自由基诱导的脂质氧化的谷胱甘肽化产物

• 批准号: 10736332
• 负责人: Robert Gerd Salomon
• 金额: $ 39.45万
• 依托单位: CASE WESTERN RESERVE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-01 至 2027-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10736332
• 关键词: Abbreviations; Acids; Address; Age related macular degeneration; Agonist; Anabolism; Autophagocytosis; Biological; Cells; Choroid; Complex; Cytokine Signaling; Detection; Development; Disease; Disease Marker; Docosahexaenoic Acids; Esters; Ethanolamines; Exhibits; Eye diseases; Free Radicals; Functional disorder; Gene Expression; Generations; Glutathione; Human; Immunoassay; In Vitro; Inflammation; Inflammatory; Interleukin-13; Knowledge; Lactones; Lecithin; Leukotriene A4; Leukotriene C4; Leukotriene D4; Leukotriene E4; Leukotriene Receptor; Leukotrienes; Light; Link; Lipids; Measures; Metabolism; Mitochondria; Modeling; Molecular; NADH; Ocular Pathology; Oxidative Stress; Oxygen; Pathogenesis; Pathologic; Pathologic Neovascularization; Pathology; Phospholipase A2; Phospholipids; Physiological; Physiology; Pilot Projects; Post-Translational Protein Processing; Production; Pyrroles; Rattus; Research; Retina; Retinal Degeneration; Retinal Edemas; Role; Scheme; Signal Pathway; Structure; Structure of retinal pigment epithelium; Testing; Therapeutic; Therapeutic Intervention; Time; Zileuton; analog; arachidonate; clinical development; cross reactivity; cysteinyl leukotriene receptor; cysteinyl-leukotriene; disabling disease; in vivo; insight; novel therapeutics; oxidation; polyunsaturated fat; protein expression; receptor; response; retinal damage; therapeutic development; transcriptome sequencing

Limited knowledge of the structures and biological activities of products generated by free radical-induced lipid oxidation in the retina is a barrier to progress in the development of clinically useful new disease markers and mechanistically informed therapeutic interventions. Understand- ing cellular responses to free radical-induced lipid oxidation products is complex owing to their diversity and ability to mimic enzymatically generated receptor agonists. Their unrecognized generation and biological activities almost certainly contribute to the mediocre efficacy of currently available therapeutic measures for age related macular degeneration (AMD). We discovered glutathione (GSH) derivatives produced from oxidatively truncated arachidonyl phospholipids that are structural and functional analogues of cysteinyl (Cys) leukotrienes (LTs). We refer to them as pseudo (ø)LTs. We will test the hypotheses that øLTs contribute to retinal pathology and physiology in a rat model of light-induced retinal degeneration. Pilot studies show that øLTs are present in human retina, are produced in vivo consequent to oxidative insult, and exhibit LT-like biological activities. Therefore, øLTs can elicit cellular responses erroneously presumed to be induced exclusively by CysLTs. Consequently, they are a confounding factor for interpreting previous studies on the involvements of LTs in retinal pathology and physiology. The proposed research will test the hypotheses that øLTs and related GSH derivatives of oxidatively truncated docosahexaenoyl phospholipids contribute to receptor-dependent inflammatory cytokine signal- ing, retinal edema, pathological neovascularization, or physiologically important initiation of retinal pigment epithelium (RPE) autophagy. The potential pathophysiological significance of these glutathionylated products of lipid oxidation lies in the possibility of their ubiquitous generation in high levels, e.g., compared to those of CysLTs. We will test the conclusion of preliminary studies that øLTs are LT receptor agonists and determine if their ability to induce the expression of IL-13 and TGF-b1 promotes LT biosynthesis. Because cross reactivity may confound the interpretation of immunoassays, we quantitate øLTs and LTs by LC-MS/MS. We will determine signaling path- ways, e.g., with RNA-Seq studies, activated by glutathionylated products of free radical-induced lipid oxidation addressing the questions: do they promote inflammation in ocular pathology or initiate autophagy in RPE cells and how do they do it? Molecular level insights into the role of oxidative stress in the pathogenesis of inflammatory eye diseases such as AMD can facilitate development of therapeutic interventions that ameliorate the progression of these common but disabling diseases.

有限的知识所产生的产品的结构和生物活性的自由 视网膜中自由基诱导的脂质氧化是临床发展的障碍， 有用的新疾病标志物和机械上知情的治疗干预。你要明白- 细胞对自由基诱导的脂质氧化产物的反应是复杂的， 多样性和模拟酶促产生的受体激动剂的能力。其未被承认的 产生和生物活动几乎肯定有助于目前的平庸功效 年龄相关性黄斑变性（AMD）的有效治疗措施。我们发现 由氧化截短的花生四烯酰磷脂产生的谷胱甘肽（GSH）衍生物， 是半胱氨酰（Cys）白三烯（LT）的结构和功能类似物。我们将它们称为 伪LT。我们将检验øLT有助于视网膜病理学的假设， 在光诱导的视网膜变性的大鼠模型中的生理学。试点研究表明，øLT是 存在于人视网膜中，是在体内产生的氧化损伤，并表现出LT样 生物活动。因此，øLT可以引起细胞反应，错误地认为是 完全由CysLT诱导。因此，它们是解释的混杂因素 以前的研究涉及的LT在视网膜病理和生理。拟议 研究将测试的假设，øLT和相关的GSH衍生物的氧化截断 二十二碳六烯酰磷脂有助于受体依赖性炎症细胞因子信号- 视网膜水肿，病理性新生血管形成，或生理上重要的视网膜病变的发生。 色素上皮（RPE）自噬。这些潜在的病理生理意义 脂质氧化的谷胱甘肽化产物存在于它们在体内普遍产生的可能性， 更高的水平，例如，与CysLT相比。我们将检验初步研究的结论 øLT是LT受体激动剂，并确定其诱导IL-13表达的能力， TGF-β 1促进LT的生物合成。因为交叉反应可能会混淆解释 我们通过LC-MS/MS定量øLT和LT。我们将确定信号传导路径- 方式，例如，通过RNA-Seq研究，由自由基诱导的谷胱甘肽化产物激活 脂质氧化处理问题：它们是否促进眼部病理学炎症 或者在RPE细胞中启动自噬，它们是如何做到的？从分子水平深入了解 氧化应激在炎症性眼病如AMD的发病机制中的作用可以促进 开发治疗干预措施，改善这些常见但 致残疾病。