Deep phenotypic and functional characterization of salt-responsive immune cells in human salt senstive hypertension using CTE-seq

使用 CTE-seq 对人类盐敏感性高血压中的盐反应性免疫细胞进行深度表型和功能表征

• 批准号: 10095170
• 负责人: Annet Kirabo
• 金额: $ 8.65万
• 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: 2021-02-01 至 2023-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10095170
• 关键词: Affect; Antibodies; Antigen-Presenting Cells; Antigens; Blood; Blood Glucose; Blood Pressure; Brain; Cardiovascular Diseases; Cardiovascular system; Cells; Cellular Indexing of Transcriptomes and Epitopes by Sequencing; Cessation of life; Cholesterol; Consumption; Data; Dendritic Cells; Dendritic cell activation; Diagnosis; Disease; Environment; Event; Excess Dietary Salt; Excretory function; Exhibits; Extracellular Fluid; Gene Expression; Genes; Hour; Human; Hypertension; Hypotension; Immune; In Vitro; Individual; Inflammation; Injury to Kidney; Intake; Interleukin-1; Interleukin-1 beta; Interleukin-6; Kidney; Laboratories; Lead; Lipid Peroxidation; Lipids; Location; Lysine; Mendelian disorder; Modeling; Myelogenous; NADPH Oxidase; Oligonucleotides; Organ; Participant; Pathogenesis; Pattern; Phenotype; Plasma; Play; Population; Production; Proteins; Pulse Pressure; Regulation; Research; Resistance; Role; Site; Sodium; Sodium Channel; Sodium Chloride; T Chain; T-Cell Receptor; T-Lymphocyte; TNF gene; Techniques; Testing; Time; Tissues; Variant; adduct; cardiovascular risk factor; cost; cost effective; cytokine; dietary salt; epithelial Na+ channel; extracellular; gain of function; high body mass index; immune activation; immunogenic; in vivo; innovation; interstitial; ketoaldehyde; loss of function; loss of function mutation; monocyte; normotensive; oxidation; personalized diagnostics; personalized medicine; response; salt intake; salt sensitive; salt sensitive hypertension; tool

Project Summary: Salt-sensitive hypertension is an independent predictor of death due to cardiovascular disease (CVD), but the mechanisms are poorly understood. Most of the research on salt-sensing mechanisms has focused on the kidney, the vasculature and the brain; however, recent studies from our laboratory have found that immune cells including antigen presenting cells (APCs) can sense sodium (Na+) and contribute to salt-induced hypertension and end organ damage through a mechanism involving increased lipid oxidation and formation of immunogenic gamma ketoaldehydes known as isolevuglandins (IsoLGs) or isoketals. Emerging evidence suggests that Na+ accumulates in the interstitium and activates immune cells but the specific tissue location including the origin, antigenic site, and final target for salt-activated immune cell in cardiovascular tissues is not known. Tools to study T cells have been rapidly expanding over the past 5 years along with increased computing capacity including single T cells sequencing of α and β chain T cell receptor spectra typing but very few studies have investigated APCs using these advanced techniques and we do not know where or how they are activated and in turn activate T cells in salt-induced CVD. We propose an innovative approach to use 5’ CITE-Seq and post hoc sequencing, to phenotype APCs in people with salt-sensitivity of blood pressure. We will couple the immune phenotype with IsoLG formation using our innovative approach to conjugate the anti- IsoLG D11 ScFv antibody sequence with an oligo and use CITE-Seq to track these important activated IsoLG-positive cells and determine if these are associated with gene expression of sodium channels. Identifying the immune cells associated with salt-sensitivity of blood pressure will have a far-ranging impact on our understanding of the pathogenesis of salt-induced hypertension and other diseases aggravated by high salt consumption.

项目概要： 盐敏感性高血压是心血管疾病死亡的独立预测因子 (CVD)，但机制知之甚少。大多数关于盐感的研究 机制集中在肾脏，血管和大脑;然而，最近的研究 我们实验室的研究人员发现，包括抗原呈递细胞（APC）在内的免疫细胞可以 感觉钠（Na+），并通过以下途径促进盐诱导的高血压和终末器官损伤 涉及脂质氧化增加和免疫原性γ形成的机制 酮醛，称为异evuglandin（IsoLG）或异缩酮。新出现的证据表明 Na+积聚在淋巴结中并激活免疫细胞，但特定的组织位置 包括盐激活免疫细胞在心血管疾病中的来源、抗原位点和最终靶点， 组织未知。研究T细胞的工具在过去5年中迅速扩大 沿着增加的计算能力，包括α和β链T细胞的单T细胞测序， 细胞受体谱分型，但很少有研究使用这些先进的APC 我们不知道它们在哪里或如何被激活，并反过来激活T细胞。 盐诱导的CVD。我们提出了一种创新的方法，使用5' CITE-Seq和post hoc 测序，表型APC在盐敏感性血压的人。我们会结合 免疫表型与IsoLG形成使用我们的创新方法，以共轭抗- IsoLG D11 ScFv抗体序列与寡核苷酸，并使用CITE-Seq来跟踪这些重要的 激活的IsoLG阳性细胞，并确定这些细胞是否与以下基因表达相关： 钠离子通道血压盐敏感性相关免疫细胞的鉴定 将对我们了解盐诱导的发病机制产生深远的影响 高血压和其他因高盐摄入而加重的疾病。