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

项目总结： 盐敏感性高血压是心血管疾病死亡的独立预测因子 (CVD)，但其机制尚不清楚。国内外对盐敏技术的研究较多 机制主要集中在肾脏、血管系统和大脑；然而，最近的研究 我们实验室的研究人员发现，包括抗原提呈细胞(APC)在内的免疫细胞可以 感知钠(Na+)并通过以下途径促进盐源性高血压和终末器官损伤 一种涉及脂质氧化增加和免疫原性γ形成的机制 被称为异丁香素(IsoLGs)或异酮的酮醛。新出现的证据表明 Na+聚集在间质中，激活免疫细胞，但在特定的组织位置 包括盐激活的心血管免疫细胞的来源、抗原部位和最终靶点 组织尚不清楚。在过去的5年里，研究T细胞的工具迅速发展 随着计算能力的增加，包括α和β链T的单T细胞测序 细胞受体光谱分型，但很少有研究使用这些先进的 技术，我们不知道它们在哪里或如何被激活，进而激活T细胞 盐诱导的脑血管病。我们提出了一种创新的方法来使用5‘Cite-Seq和POST HOC 对血压盐敏感者的APC表型进行测序。我们会结对的 使用我们的创新方法结合抗-HBs的免疫表型与IsoLG形成 IsoLG D11单链抗体序列与寡核苷酸结合，并使用Cite-Seq追踪这些重要的 激活IsoLG阳性细胞并确定它们是否与基因表达有关 钠离子通道。识别与血压盐敏感性相关的免疫细胞 将对我们理解盐中毒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.