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: 10255052
• 负责人: Justin Pieter Van Beusecum
• 金额: --
• 依托单位: VETERANS HEALTH ADMINISTRATION
• 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/10255052
• 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 reduction; blood pressure regulation; cell growth; combat; curative treatments; cytokine; exposed human population; genetic predictors; human subject; 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.

项目总结 高血压是中风、心肌梗死、心脏病等疾病发病率和死亡率的主要原因 失败，以及美国退伍军人的慢性肾脏疾病。尽管重要的是 在血压控制方面，高血压的发病机制仍知之甚少。 最近，一项新的观察揭示了潜在的树突状细胞(DC)可能参与 人类高血压。利用单细胞核糖核酸(RNA)测序，一种新的DC亚群 AX1和Siglec-6在正常健康受试者中的表面表达。他们展示了 Axl+Siglec-6+DC(AS DC)可以有效地促进T细胞的增殖并产生大量的 促炎细胞因子。根据之前发表的研究和初步数据，我建议 高血压导致内皮源性生长抑制特异性6(Gas6)的释放，Gas6激活 AXL对树突状细胞导致炎症及其相关的终末器官损伤。在目标1中，我将测试 高血压刺激导致Gas6/Axl依赖的炎症体激活的假说 由AS DC中的活性氧物种(ROS)决定。在这个目标中，我将人类CD14+单核细胞暴露于 Gas6或高血压内皮细胞拉伸和评估炎症小体激活，ROS产生， 流式细胞术检测高活性异丙肾上腺素加合物的形成。我也会 用2-羟色胺清除异丙基葡萄糖，研究异丙基葡萄糖在炎症体激活中的作用 人单核细胞暴露于高血压刺激时的羟基苯胺胺(2-HOBA) 用流式细胞仪检测细胞因子的产生。最后，我会检讨区议会对 血压正常和高血压的受试者无偏的细胞索引 转录本和表位测序(引用序列)。我预测清除ROS和/或IsoLGs 在暴露于Gas6或内皮细胞时，拉伸可防止炎症体激活和促进 AS DC释放炎性细胞因子。在目标2中，我将测试AXL依赖的假设 树突状细胞中的信号促进高血压及其相关的终末器官损伤。我将使用老鼠，在其中 我已经专门在DC(Axlfl/fl和AxlCD11cCre)中删除了Axl。我将研究Axl在DC中的体内作用 用放射遥测和流式细胞仪检测高血压患者的血压。接下来，我将调查AXL在 流式细胞仪检测DCs体外对炎性小体激活和T细胞增殖的影响我预测基因 AXL的缺失将防止高血压、DC激活及其相关的炎症。在《目标3》中，我会 检验Gas6促进DC激活、高血压及其相关炎症的假设 我将使用Gas6基因缺失的小鼠(GAS6WT/WT和Gas6-/-)。我会检查体内的 Gas6在骨髓移植研究中的作用和评估DC激活和 应用流式细胞仪和放射遥测技术研究高血压的进展。我将从基因上删除Gas6 并用流式细胞仪检测DC的活化状态和细胞因子的产生。我 预测Gas6基因缺失将防止高血压和免疫细胞的激活。这些 研究将促进我们对DC在人类高血压发展中所扮演的角色的理解 将为美联航治疗人类心血管疾病提供新的治疗方向 州退伍军人。