A Role of GAS6/Axl Signaling in the Development of Essential Hypertension

GAS6/Axl 信号传导在原发性高血压发展中的作用

• 批准号: 10447079
• 负责人: Justin Pieter Van Beusecum
• 金额: --
• 依托单位: RALPH H JOHNSON VA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-07-01 至 2026-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10447079
• 关键词: Adult; Affect; Age; American; Angiotensin II; Animal Model; Anti-Inflammatory Agents; Atherosclerosis; Award; Blood Pressure; Blood Vessels; Bone Marrow Transplantation; CD14 gene; Cardiovascular Diseases; Cause of Death; Cells; Cellular Indexing of Transcriptomes and Epitopes by Sequencing; Chronic Kidney Failure; Comb animal structure; Cytometry; Data; Dendritic Cells; Dendritic cell activation; Development; Diagnosis; Disease; Endothelial Cells; Endothelium; Epidemic; Essential Hypertension; Experimental Models; Flow Cytometry; Genetic; Genetic Techniques; Growth; Guidelines; Health; Heart failure; Human; Hypertension; In Vitro; Inflammasome; Inflammation; Inflammatory; Kidney; Lead; Ligands; Light; Malignant Neoplasms; Mechanics; Mediating; Methods; Molecular Genetics; Morbidity - disease rate; Mus; Myocardial Infarction; Organ; Oxidants; Oxidative Stress; Pathogenesis; Pharmacology; Physiological; Play; Production; Publishing; RNA; Reactive Oxygen Species; Receptor Protein-Tyrosine Kinases; Risk Factors; Role; Signal Transduction; Stimulus; Stretching; Stroke; Surface; T-Cell Proliferation; Testing; United States; Veterans; adduct; base; blood pressure control; blood pressure reduction; cell growth; combat; curative treatments; cytokine; exposed human population; genetic predictors; human subject; hypertensive; immune activation; improved; in vivo; military veteran; monocyte; mortality; new therapeutic target; normotensive; novel; novel therapeutics; prevent; receptor; renal damage; response; sialic acid binding Ig-like lectin; therapy outcome; translational approach

PROJECT SUMMARY Hypertension is the leading cause of morbidity and mortality from stroke, myocardial infarction, heart failure, and chronic kidney disease amongst the United States Veterans. Despite the importance of blood pressure control, the pathogenesis of essential hypertension remains poorly understood. Recently, a new observation has shed light onto potential dendritic cells (DCs) that may be involved in human hypertension. Using single-cell ribonucleic acid (RNA) sequencing, a new subset of DCs has been described with surface expression of Axl and Siglec-6 in normal health subjects. They showed that Axl+ Siglec-6+ DCs (AS DCs) can potently drive T cell proliferation and produce large amounts of pro-inflammatory cytokines. Based on previously published studies and preliminary data, I propose that hypertension leads to the release of endothelial-derived growth arrest specific 6 (GAS6) that activates Axl on DCs leading to inflammation and its associated end organ damage. In Aim 1, I will test the hypothesis that hypertensive stimuli lead to GAS6/Axl-dependent inflammasome activation mediated by reactive oxygen species (ROS) in AS DCs. In this aim, I will expose human CD14+ monocytes to GAS6 or hypertensive endothelial cell stretch and assess inflammasome activation, ROS production, and the formation of the highly reactive Isolevuglandin (isoLG)-adducts by flow cytometry. I will also examine the role of isoLGs on inflammasome activation by scavenging isoLGs using 2- hydroxybenzilamine (2-HOBA) during exposure of human monocytes to hypertensive stimuli and assess cytokine production by flow cytometry. Lastly, I will examine the response of AS DCs from normotensive and hypertensive human subjects in an unbiased fashion by cellular indexing of transcriptomes and epitopes by sequencing (CITE-seq). I predict that scavenging ROS and/or isoLGs during exposure of GAS6 or endothelial cell stretch will prevent inflammasome activation and pro- inflammatory cytokine release by AS DCs. In Aim 2, I will test the hypothesis that Axl-dependent signaling in DCs promotes hypertension and its associated end-organ damage. I will use mice in which I have deleted Axl specifically in DCs (Axlfl/fl and AxlCD11cCre). I will examine the in vivo role of Axl in DCs during hypertension by radiotelemetry and flow cytometry. Next, I will investigate the deletion of Axl in DCs on inflammasome activation and T cell proliferation in vitro by flow cytometry. I predict that genetic deletion of Axl will prevent hypertension, DC activation, and its associated inflammation. In Aim 3, I will test the hypothesis that GAS6 promotes DC activation, hypertension, and its associated inflammation I will use mice that have genetic deletion for GAS6 (GAS6WT/WT and GAS6-/-). I will examine the in vivo role of GAS6 by performing bone marrow transplantation studies and assess DC activation and development of hypertension by flow cytometry and radiotelemetry. I will genetically delete GAS6 from human endothelial cells and assess DC activation status and cytokine production by flow cytometry. I predict that genetic deletion of GAS6 will prevent hypertension and activation of immune cells. These studies will advance our understanding of the role DCs play in the development of human hypertension and will provide new therapeutic directions for the treatment of human cardiovascular disease for United States Veterans.

项目摘要 高血压是中风，心肌梗塞，心脏的发病率和死亡率的主要原因 美国退伍军人的失败和慢性肾脏疾病。尽管很重要 血压控制，基本高血压的发病机理仍然了解不足。 最近，一种新的观察结果使可能与潜在的树突状细胞（DC）相关 人类高血压。使用单细胞核糖核酸（RNA）测序，一个新的DCS子集具有 在正常健康受试者中用AXL和SIGLEC-6的表面表达描述。他们表明 AXL+ SIGLEC-6+ DC（作为DC）可以有效地驱动T细胞增殖并产生大量 促炎性细胞因子。根据先前发表的研究和初步数据，我建议 高血压导致释放激活的内皮衍生生长阻滞特异性6（GAS6） DC上的AXL导致炎症及其相关的最终器官损伤。在AIM 1中，我将测试 假设高血压刺激导致GAS6/AXL依赖性炎症体激活介导 通过AS DC中的活性氧（ROS）。在此目标中，我将使人CD14+单核细胞暴露于 GAS6或高血压内皮细胞拉伸并评估炎症体激活，ROS产生， 以及由流式细胞仪形成高度反应性异甲列蛋白（ISLEG） - 添加。我也会 通过使用2-清除分离犬，检查了分离菌在炎性体激活中的作用 在人类单核细胞暴露于高血压刺激和 通过流式细胞仪评估细胞因子的产生。最后，我将检查AS DC的响应 通过蜂窝索引以公正的方式进行异常的和高血压的人类受试者 通过测序（cite-seq）进行转录组和表位。我预测清除ROS和/或隔离 在暴露于气体或内皮细胞拉伸期间，将防止炎症体激活和促进 AS DCS释放炎症细胞因子。在AIM 2中，我将测试依赖AXL的假设 DC中的信号促进高血压及其相关的最终器官损伤。我会用老鼠 我在DC（AXLFL/FL和AXLCD11CCRE）中删除了AXL。我将检查AXL在DC中的体内作用 在辐射电子测定和流式细胞术中高血压期间。接下来，我将研究AXL的删除 通过流式细胞仪在体外炎性体激活和T细胞增殖上的DC。我预测遗传 AXL的缺失将防止高血压，直流激活及其相关的炎症。在AIM 3中，我会 测试GAS6促进直流激活，高血压及其相关炎症的假设 我将使用具有遗传缺失的小鼠（gas6wt/wt和gas6 - / - ）。我将检查体内 通过进行骨髓移植研究并评估DC激活和 通过流式细胞仪和放射性终极测定的高血压发展。我会从基因删除GAS6中 人体内皮细胞并通过流式细胞仪评估DC激活状态和细胞因子产生。我 预测GAS6的遗传缺失将防止免疫细胞的高血压和激活。这些 研究将促进我们对DCS在人类高血压发展中所扮演的作用的理解 并将为治疗人类心血管疾病的治疗提供新的治疗方向 州退伍军人。