Single cell analysis of gene expression in human vascular cells

人类血管细胞基因表达的单细胞分析

• 批准号: 9810454
• 负责人: RAJAT M GUPTA
• 金额: $ 8.95万
• 依托单位: BRIGHAM AND WOMEN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-08-01 至 2021-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9810454
• 关键词: 6p24; Architecture; Arterial Fatty Streak; Arteries; Atherosclerosis; Atlases; Biological; Biological Process; Biology; Blood Vessels; Cells; Cessation of life; Chromosomes; Clinical; Coronary Arteriosclerosis; Data; Detection; Development; Disease; Disease Progression; Dissection; Distal; Endothelial Cells; Endothelin-1; Foundations; Funding; Future; Gene Expression Profile; Gene Expression Profiling; Genetic; Genetic Transcription; Genetic Variation; Genomics; Goals; Grant; Heterogeneity; Hospitals; Human; Human Genetics; Integrins; Methods; Migraine; Mus; Nuclear; Outcome; Pathway interactions; Patients; Population; Prevention; RNA; Regulator Genes; Research; Resources; Role; Stroke; Techniques; Temporal Arteries; Tissues; Training; United States; Untranslated RNA; Variant; Vascular Diseases; Vasoconstrictor Agents; Woman; Work; base; cell type; disability; disorder risk; epigenomics; experimental study; functional genomics; genetic variant; genome editing; genome wide association study; human RNA sequencing; lipid transport; medical schools; new therapeutic target; preservation; single cell analysis; single-cell RNA sequencing; skills; transcriptome sequencing

Project Summary In my clinical training, I repeatedly saw the devastating effects of vascular disease on patients in the prime of their lives. Vascular diseases such as coronary artery disease (CAD), stroke, arterial dissection, and migraine headache combine to cause over half the death and disability in the United States. To eradicate vascular disease it will be important to develop new treatments that target the arterial cells where the disease begins. My K08- funded research has been to identify these new pathways using human genetic variation as a guide. The loci associated with multiple vascular diseases have recently been identified through genome-wide association studies (GWAS). These loci represent new therapeutic targets, but their biological mechanisms remain largely unexplored. I have identified the mechanism by which a vascular disease associated variant on chromosome 6p24 distally regulates endothelin-1 expression. This regulatory effect of the non-coding variant is vascular tissue specific, and largely seen in endothelial cells. Whether other cells in the blood vessel are responsible for other gene regulatory effects for this locus remains unknown. For this and other vascular disease-associated loci it will be important to characterize the gene regulatory effects in the relevant cell type or cellular subpopulation. New methods in single cell RNA-sequencing all the identification of the full set of cells in the arterial wall. Droplet-based single cell RNA-sequencing allows for analysis of thousands of cells and detection of rare cellular subpopulations. Using this method, I have identified three distinct subpopulations of endothelial cells with transcriptional signatures that suggest functional specialization. This R03 application seeks to extend this finding to human vascular tissue. The goals of this proposal are to determine the best method for single cell RNA-sequencing and to determine the markers of endothelial cell heterogeneity in human vascular tissue. This will provide an atlas of cell types based on transcriptional signature in vascular tissue and serve as a foundational resource for future functional genomics experiments. With a better understanding of the cell types and gene expression signatures in the arterial wall it will be possible to identify new biological pathways for the treatment and prevention of vascular disease.

项目概要 在我的临床培训中，我多次看到血管疾病对身体的破坏性影响 处于人生巅峰的患者。血管疾病，例如冠状动脉疾病 （CAD）、中风、动脉夹层和偏头痛共同导致超过一半的 美国的死亡和残疾。根除血管疾病很重要 开发针对疾病开始的动脉细胞的新疗法。我的K08- 资助的研究是利用人类遗传变异来识别这些新途径 作为指导。最近发现与多种血管疾病相关的位点 通过全基因组关联研究（GWAS）确定。这些基因座代表新的 治疗靶点，但其生物学机制在很大程度上仍未被探索。我有 确定了血管疾病相关染色体变异的机制 6p24 远端调节内皮素-1 表达。非编码的这种调节作用 变异是血管组织特异性的，主要见于内皮细胞。是否其他 血管中的细胞负责该位点的其他基因调节作用 仍然未知。对于这个和其他血管疾病相关位点来说，这很重要 表征相关细胞类型或细胞中的基因调控效应 亚人群。单细胞RNA测序的新方法所有鉴定 动脉壁中的全套细胞。基于液滴的单细胞 RNA 测序可实现 分析数千个细胞并检测稀有细胞亚群。使用这个 方法，我已经鉴定出三种不同的内皮细胞亚群 表明功能专业化的转录特征。这个R03应用程序 试图将这一发现扩展到人体血管组织。该提案的目标是 确定单细胞 RNA 测序的最佳方法并确定标记 人类血管组织中内皮细胞异质性的研究。这将提供一个地图集 基于血管组织中转录特征的细胞类型，并作为 未来功能基因组学实验的基础资源。有了更好的 了解动脉壁中的细胞类型和基因表达特征 可能找到治疗和预防疾病的新生物学途径 血管疾病。