Anti-inflammatory actions of a novel hemin-induced electrophile in Sickle Cell Disease

新型血红素诱导的亲电子试剂在镰状细胞病中的抗炎作用

• 批准号: 10402907
• 负责人: Dario A Vitturi
• 金额: $ 0.37万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-05-15 至 2022-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10402907
• 关键词: Acute; Adhesiveness; Affect; African American population; Animals; Anti-Inflammatory Agents; Antioxidants; Attenuated; Award; Biochemical; Biological; Blood Platelets; Blood Vessels; Cell Culture System; Chemistry; Child; Chronic; Cysteine; Data; Deamination; Development; Disease; Drug or chemical Tissue Distribution; Endothelium; Erythrocytes; Exhibits; Fatty Acids; Fluorescence; Foundations; Functional disorder; Future; Gene Expression; Generations; Genes; Genetic; Goals; Health; Hematological Disease; Hematology; Heme; Hemin; Hemoglobin; Hemolytic Anemia; High Pressure Liquid Chromatography; Hispanic Americans; Human; In Vitro; Individual; Inflammasome; Inflammation; Inflammation Mediators; Inflammatory; Infusion procedures; Injury; Kidney Diseases; Knowledge; Kynurenine; Leukocytes; Life; Life Expectancy; Liver; Lung; Mass Spectrum Analysis; Mediator of activation protein; Metabolism; Microscopy; Mission; Morbidity - disease rate; Mus; Myeloid Cells; Nature; Organ; Oxidative Stress; Pain; Pathology; Pathway interactions; Patients; Penetrance; Persons; Plasma; Primary Cell Cultures; Process; Public Health; Quality of life; Recurrence; Research; Role; Sampling; Secondary to; Sickle Cell; Sickle Cell Anemia; Signal Transduction; Solid; Sulfhydryl Compounds; TLR4 gene; Testing; Therapeutic; Tissues; Transgenic Mice; Transgenic Organisms; Translational Research; Translations; Tryptophan 2,3 Dioxygenase; Tryptophanase; Tubular formation; United States National Institutes of Health; Up-Regulation; Urine; analytical tool; antioxidant enzyme; clinically relevant; design; disability; disease heterogeneity; effective therapy; genetic approach; genetic regulatory protein; heme oxygenase-1; hemoglobin AA; human disease; immune activation; improved; in vivo; individualized medicine; intravital microscopy; metabolomics; mouse model; neutrophil; novel; organ injury; prevent; prognostic; psychologic; response; stem; tissue injury; treatment strategy; urinary; vaso-occlusive crisis

Sickle Cell Disease (SCD) is the most prevalent genetic blood disorder in the US affecting an estimated 1 in 365 African Americans and 1 in 16,300 Hispanic Americans. SCD is associated with decreased life expectancy and significant physical and psychological morbidity due to complications arising from recurrent vaso-occlusion and hemolytic anemia. The activation of TLR4 by hemoglobin-derived products is a central component of a pernicious inflammatory cascade that involves endothelial activation, formation of multicellular platelet/neutrophil/erythrocyte aggregates and oxidative stress. The effects of SCD on kynurenine metabolism and in particular, on the generation of a novel bioactive kynurenine-derived metabolite were hitherto unappreciated. The bioactive kynurenine-derived mediator discovered herein activates Nrf2-dependent expression of antioxidant enzymes and inhibits NF-κB regulated inflammatory signaling as well as abrogates NLRP3 inflammasome activity. Formation of the bioactive kynurenine metabolite is upregulated in steady state in both mouse models and human SCD patients, and preliminary data suggests that levels are further increased by hemin, a critical mediator of vaso-occlusive crises (VOC) and organ injury. By inhibiting NF-κB- dependent responses and downregulating the NLRP3 inflammasome, this novel kynurenine-derived compound has the potential to act as an endogenous anti-inflammatory mediator capable of regulating endothelial, platelet and immune cell activation to attenuate VOC. Furthermore, Nrf2 activation is known to be protective in SCD via inhibition of pro-inflammatory signaling, upregulation of the heme-catabolizing enzyme HO-1, and by increasing endogenous antioxidant defenses that scavenge reactive species. By utilizing relevant primary cell culture systems derived from animals bearing key pathway deletions, a clinically relevant SCD transgenic mouse model, highly specific LC-MS/MS strategies and state-of-the-art quantitative fluorescence intravital lung microscopy, the research plan will establish the mechanisms behind the anti-inflammatory actions of the bioactive kynurenine metabolite and its potential to attenuate vaso-occlusion in vivo. The present application is built on solid biochemical foundations, and applies critical biological and analytical tools to successfully transition from in vitro chemistry to translational science with the ultimate objective of positively impacting human health. The knowledge gained from this research plan will be of pivotal importance for generating solid preliminary for future R01 applications aimed at better defining the pathophysiological role of the kynurenine- derived bioactive metabolite in SCD as well as in other hematological and non-hematological conditions in which chronic inflammation, oxidative stress and ischemic organ injury are involved.

镰状细胞病（SCD）是美国最常见的遗传性血液疾病，影响估计 每 365 名非洲裔美国人中就有 1 人，16,300 名西班牙裔美国人中就有 1 人。 SCD 与寿命缩短有关 由于复发性并发症引起的预期和显着的身体和心理发病率 血管闭塞和溶血性贫血。血红蛋白衍生产物激活 TLR4 是一个重要的机制。 恶性炎症级联的组成部分，涉及内皮激活、多细胞形成 血小板/中性粒细胞/红细胞聚集和氧化应激。 SCD对犬尿氨酸代谢的影响 特别是，迄今为止，关于新型生物活性犬尿氨酸衍生代谢物的生成 不被欣赏。本文发现的生物活性犬尿氨酸衍生介质激活 Nrf2 依赖性 抗氧化酶的表达并抑制 NF-κB 调节的炎症信号传导以及消除 NLRP3 炎症小体活性。生物活性犬尿氨酸代谢物的形成在稳态下上调 在小鼠模型和人类 SCD 患者中，初步数据表明该水平进一步 血红素会增加血红素，血红素是血管闭塞危机 (VOC) 和器官损伤的关键介质。通过抑制 NF-κB- 依赖性反应并下调 NLRP3 炎症小体（这种新型犬尿氨酸衍生化合物） 具有作为内源性抗炎介质的潜力，能够调节内皮细胞， 血小板和免疫细胞激活以减弱 VOC。此外，已知 Nrf2 激活具有保护作用 SCD 通过抑制促炎信号传导、上调血红素分解代谢酶 HO-1 以及通过 增加清除活性物质的内源性抗氧化防御。利用相关原代细胞 源自具有关键通路缺失的动物的培养系统，这是一种临床相关的 SCD 转基因 小鼠模型、高度特异性的 LC-MS/MS 策略和最先进的定量荧光活体肺 显微镜下，该研究计划将建立抗炎作用背后的机制 生物活性犬尿氨酸代谢物及其减轻体内血管闭塞的潜力。本申请是 建立在坚实的生化基础上，并应用关键的生物和分析工具成功地 从体外化学到转化科学的转变，最终目标是产生积极影响 人类健康。从该研究计划中获得的知识对于生成固体至关重要 为未来的 R01 应用做准备，旨在更好地定义犬尿氨酸的病理生理作用- SCD 以及其他血液学和非血液学病症中衍生的生物活性代谢物 其中涉及慢性炎症、氧化应激和缺血性器官损伤。