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

项目摘要 高血压是一个主要的全球健康问题。仅在美国，三分之一的成年人患有高血压， 到60岁时，这一比例上升到三分之二。高血压是导致高血压病发病率和死亡率的关键因素， 中风、心肌梗塞、心力衰竭、外周血管疾病和肾衰竭/透析。不幸的是， 尽管目前的治疗，血压仍然控制不佳，在约50%的个人， 这种疾病。即使血压得到合理控制，高血压也与血压升高有关。 心血管风险我们的实验室和其他实验室有新的证据表明，这种剩余风险和最终- 高血压的器官效应主要由炎症介导。例如，缺乏T细胞的小鼠 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的 高血压和降低的对高血压刺激的反应的终末器官功能障碍。但是大部分这些 在实验动物中进行了研究，特别是啮齿动物， 炎症在人类高血压中的作用此外，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，该提案的第一个目标是利用尖端的单细胞 多重质谱飞行时间（CyTOF）方法来分析外周血中的免疫细胞， 正常血压和高血压的人，以确定独特的和可能罕见的亚群，改变了在 人类高血压在质谱细胞术中，细胞外和细胞内靶标的抗体缀合至 由质谱仪检测的稀土金属同位素。由于肥胖和衰老是错综复杂的 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， 年龄（35至75岁），以确定观察到的免疫谱中的高血压变化与 肥胖和衰老。第二个目标是通过流式细胞术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细胞的深度测序对其进行分选和进一步表征， 在T淋巴细胞的情况下细胞受体。绘制高血压，肥胖， 和衰老有望导致新的诊断，预后和治疗策略，以治疗人类 高血压和限制相关的终末器官损害，从这种慢性，广泛的疾病， 引起全身免疫抑制。

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.