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

项目摘要： 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。

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.