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.

项目概要 高血压是全球主要的健康问题。仅在美国，三分之一的成年人患有高血压和 到 60 岁时，这一数字将上升至三分之二。高血压是导致以下疾病发病和死亡的一个关键因素： 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， 尽管目前正在接受治疗，但约 50% 的患者血压仍控制不佳 这种病。即使血压得到合理控制，高血压仍与血压升高有关。 心血管风险。我们的实验室和其他实验室的新证据表明，这种残余风险和最终- 高血压对器官的影响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/巨噬细胞或多种促炎细胞因子之一表现出钝化 高血压并减少对高血压刺激的终末器官功能障碍。然而这些大多数 已经在实验动物，特别是啮齿类动物中进行了研究，有限的研究检验了 炎症对人类高血压的影响。此外，人类的整体甚至部分免疫抑制 并非没有风险。为了解决这个问题，该提案的首要目标是利用尖端的单电池 多重质谱流式细胞仪飞行时间 (CyTOF) 方法分析外周血中的免疫细胞 血压正常和高血压的人类，以确定独特且可能罕见的亚群，这些亚群在 人类高血压。在质谱流式分析中，细胞外和细胞内靶标的抗体与 由质谱仪检测到的稀土金属同位素。由于肥胖和衰老错综复杂 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和 年龄（35 至 75 岁）以确定观察到的免疫特征中高血压变化的相关程度 与肥胖和衰老有关。第二个目标是通过流动分离这些新的或改变的细胞群 通过 RNA 测序、细胞因子产生和/或 T 的深度测序来对它们进行分类和进一步表征 T 淋巴细胞的细胞受体。绘制高血压、肥胖症的免疫学图谱 衰老有望带来新的诊断、预后和治疗策略来治疗人类 高血压并限制这种慢性、广泛疾病造成的相关终末器官损伤，而无需 引起全身免疫抑制。