Immunophenotyping of Human Hypertension Using Single Cell Multiplex Mass Cytometry to Identify Novel Therapeutic Targets

使用单细胞多重质谱流式细胞仪对人类高血压进行免疫表型分析，以确定新的治疗靶点

• 批准号: 10000699
• 负责人: Meenakshi Swaminathan Madhur
• 金额: $ 47.1万
• 依托单位: VANDERBILT UNIVERSITY 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 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10000699
• 关键词: Address; Adult; Age; Aging; Animals; Antibodies; B-Lymphocytes; Blood Pressure; Body mass index; Cells; Chronic; Cytometry; Dialysis procedure; Disease; Exhibits; Functional disorder; Goals; Heart; Heart failure; Human; Hypertension; Immune; Immune system; Immunologics; Immunophenotyping; Immunosuppression; Individual; Inflammation; Inflammatory; Isotopes; Kidney Diseases; Kidney Failure; Lead; Mediating; Morbidity - disease rate; Mus; Myocardial Infarction; Obesity; Organ; Peripheral Vascular Diseases; Population; Production; Public Health; Rare Earth Metals; Residual state; Risk; Rodent; Sorting - Cell Movement; Stimulus; Stroke; T cell receptor repertoire sequencing; T-Lymphocyte; Therapeutic; Time; Widespread Disease; cardiovascular risk factor; cytokine; deep sequencing; extracellular; global health; macrophage; mass spectrometer; monocyte; mortality; new therapeutic target; normotensive; novel; novel diagnostics; novel strategies; peripheral blood; prognostic; response; study population; transcriptome sequencing

PROJECT SUMMARY Hypertension is a major global health concern. In the U.S. alone, one-third of all adults have hypertension and this rises to two-thirds by the age of 60. Hypertension is a key contributor to morbidity and mortality from stroke, myocardial infarction, heart failure, peripheral vascular disease, and renal failure/dialysis. Unfortunately, despite current treatment, blood pressure remains poorly controlled in approximately 50% of individuals with this disease. Even when blood pressure is reasonably controlled, hypertension is associated with increased cardiovascular risk. There is emerging evidence from our lab and others that this residual risk and the end- organ effects of hypertension are largely mediated by inflammation. For example, mice that are deficient in T cells, B cells, monocytes/macrophages, or one of a number of pro-inflammatory cytokines exhibit blunted hypertension and reduced end-organ dysfunction in response to hypertensive stimuli. Yet most of these studies have been conducted in experimental animals, particularly rodents, with limited studies examining the effect of inflammation in human hypertension. Moreover, global or even partial immunosuppression in humans is not without risk. To address this problem, the first goal of this proposal is to utilize a cutting-edge single cell multiplex mass cytometry time of flight (CyTOF) approach to profile the immune cells in the peripheral blood of normotensive and hypertensive humans to identify unique and possibly rare subpopulations that are altered in human hypertension. In mass cytometry, antibodies to extracellular and intracellular targets are conjugated to rare earth metal isotopes that are detected by a mass spectrometer. As obesity and aging are intricately associated with hypertension, our study population will include people with a range of body mass indices and age (35 to 75) to determine the extent to which observed hypertensive changes in immune profiles correlate with obesity and aging. The second goal is to isolate these novel or altered cell populations through flow sorting and further characterize them by RNA-sequencing, cytokine production, and/or deep sequencing of T cell receptors in the case of T lymphocytes. Mapping the immunological landscape of hypertension, obesity, and aging promises to lead to new diagnostic, prognostic, and therapeutic strategies to treat human hypertension and limit the associated end-organ damage from this chronic, widespread disease without inducing global immunosuppression.

项目摘要 高血压是全球主要的健康问题。仅在美国，所有成年人中有三分之一患有高血压和 到60岁时，这将上升到三分之二。高血压是导致发病率和死亡率的关键 中风，心肌梗塞，心力衰竭，周围血管疾病和肾衰竭/透析。很遗憾， 尽管目前治疗，但在大约50％的患有 这种疾病。即使血压得到合理控制，高血压也会增加 心血管风险。我们实验室和其他人有新的证据表明，这种残留风险和最终的风险 高血压的器官作用主要是由炎症介导的。例如，缺乏T的小鼠 细胞，B细胞，单核细胞/巨噬细胞或许多促炎细胞因子中的一种表现出钝的 响应高血压刺激，高血压和末端器官功能障碍。然而大多数 在实验动物，尤其是啮齿动物的研究中进行了研究，研究有限的研究 炎症在人类高血压中的影响。而且，人类的全球甚至部分免疫抑制 并非没有风险。为了解决这个问题，该提案的第一个目标是利用尖端的单个单元格 飞行的多重质量细胞仪（Cytof）方法，以介绍外周血中的免疫细胞 正常的和高血压的人类，以识别有所改变的独特且可能是罕见的亚群 人类高血压。在质量细胞仪中，对细胞外和细胞内靶标的抗体被缀合 质谱仪检测到的稀土金属同位素。因为肥胖和衰老很复杂 与高血压相关，我们的研究人群将包括具有一系列体重指数的人和 年龄（35至75），以确定观察到免疫特征的高血压变化相关的程度 肥胖和衰老。第二个目标是通过流量隔离这些新颖或改变的细胞种群 通过RNA测序，细胞因子产生和/或T的深度测序来分类和进一步表征它们 在T淋巴细胞的情况下。绘制高血压，肥胖， 衰老有望导致新的诊断，预后和治疗策略来治疗人类 高血压并限制这种慢性，广泛疾病的相关最终器官损害 诱导全球免疫抑制。