From genotype to phenotype in a GWAS locus: the role of REST in atherosclerosis

GWAS 位点从基因型到表型：REST 在动脉粥样硬化中的作用

基本信息

批准号：
10570469
负责人：
Marios Arvanitis
金额：
$ 17.17万
依托单位：
JOHNS HOPKINS UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-02-01 至 2028-01-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10570469
关键词：
Advisory Committees Affect Alleles Aorta Arteries Atherosclerosis Award Bayesian Method Binding Sites Biology Blood Vessels CRISPR/Cas technology Cardiovascular Diseases Cause of Death Cellular Assay Cessation of life Chromosome 4 Clinical Clustered Regularly Interspaced Short Palindromic Repeats Coronary Arteriosclerosis Coronary heart disease DNA Data Data Set Dependovirus Development Disease Distal Distant Endothelial Cells Endothelium Enhancers Enterobacteria phage P1 Cre recombinase Epigenetic Process Exposure to Funding Future Gene Silencing Gene Targeting Genes Genetic Genetic Risk Genomics Genotype Heritability High Fat Diet Human Human Genetics Immunofluorescence Immunologic In Vitro Individual Injections Investigation K-Series Research Career Programs Knock-out Knockout Mice Laboratories LoxP-flanked allele Matrix Metalloproteinases Mediating Mentors Mesenchymal Methods Multiomic Data Mus NOS3 gene Neural Cell Adhesion Molecule L1 PECAM1 gene Pathway Analysis Pathway interactions Permeability Phenotype Physicians Population Postdoctoral Fellow Proteins Public Health Publishing Research Research Personnel Rest Role SOX18 gene Scientist Site Small Interfering RNA Tamoxifen Testing Time Tissues Translating United States Untranslated RNA Variant Vascular Endothelial Cell Vascular Endothelium Woman Work angiogenesis cadherin 5 cardiovascular disorder risk causal variant chromatin immunoprecipitation data integration design disorder risk effective therapy experimental study factor A genetic analysis genetic approach genetic variant genome editing genome wide association study genomic locus human disease human stem cells in vivo inducible Cre insight men migration mouse model multidisciplinary novel novel therapeutics overexpression programs response risk variant single-cell RNA sequencing skills stem cell differentiation success transcription factor transcription factor USF transcriptome transcriptome sequencing

项目摘要

Project Summary This K08 mentored clinical scientist research career development award is a five-year program designed to facilitate Dr. Marios Arvanitis’ (PI) development into an independent physician-investigator in vascular genetics. Atherosclerotic cardiovascular disease (ASCVD) is a major public health burden that accounts for over 600,000 deaths in the United States each year. ASCVD is highly heritable and genome-wide association studies have discovered many candidate genomic loci that increase the risk of the disease in the population, thereby providing a window to novel therapies. However, most genomic risk loci for ASCVD remain unexplored in terms of how they lead to disease risk. Previously published work by the PI has focused on the mechanistic interpretation of genomic risk loci for cardiovascular disease, including the development of a novel Bayesian method, called CAFEH, to prioritize the target tissue and genes in genomic loci. Our preliminary analyses of the genetic underpinnings of ASCVD reveal that endothelial cells are enriched for ASCVD heritability, and we have used those methods to prioritize a chromosome 4 locus that is predicted to affect ASCVD risk via altering the expression of the RE-1 silencing transcription factor (REST) gene in endothelial cells. This K08 project will explore the regulatory mechanisms via which the REST locus and gene influence the development of atherosclerosis. Aim 1 will employ CRISPR-Cas9 editing in human stem cells which will then be differentiated into endothelial cells to identify the causal variants and the upstream transcription factors that mediate the association in the 4q12 coronary disease GWAS locus. Aim 2 will define distal genes and pathways affected by REST in the endothelium and investigate their cellular consequences, starting with evaluating the role of REST in endothelial to mesenchymal transition. Aim 3 will use a tamoxifen inducible endothelial specific Rest knock-out mouse model to evaluate the in vivo effects of Rest in the endothelium and atherosclerosis. The success of this project is guaranteed by the support of a multidisciplinary mentoring team including a vascular biologist (Dr. Charles Lowenstein), a computational biologist (Dr. Alexis Battle), and a functional genetics expert (Dr. Andrew McCallion), along with an advisory committee of experts in vascular biology, stem cell differentiation and atherosclerosis (Dr. Harry Dietz, Dr. Chulan Kwon and Dr. Thomas Quertermous). This award period will help the PI boost their genomics skills, acquire new wet lab skills and generate preliminary data to successfully compete for R01 funding in order to translate the genetic insights into novel mechanisms for ASCVD.

项目摘要这个K08指导的临床科学家研究职业发展奖是一个为期五年的计划促进Marios Arvanitis博士（PI）发育成为血管遗传学中独立的物理评估剂。动脉粥样硬化心血管疾病（ASCVD）是一种主要的公共卫生烧伤美国每年有60万人死亡。 ASCVD是高度遗传和全基因组的关联研究已经发现了许多候选基因组基因局，从而增加了人群疾病的风险提供新疗法的窗口。但是，ASCVD的大多数基因组风险基因座在它们如何导致疾病风险。 PI先前发表的工作重点是基因组风险基因座的机械解释对于心血管疾病，包括开发一种名为Cafeh的新型贝叶斯方法，以优先考虑基因组基因座中的靶组织和基因。我们对ASCVD遗传基础的初步分析揭示内皮细胞具有富含ASCVD的遗传力，我们已经使用这些方法优先考虑预测通过改变RE-1沉默的表达会影响ASCVD风险的4个基因座内皮细胞中的转录因子（静止）基因。这个K08项目将通过其余的基因座和基因会影响动脉粥样硬化的发展。 AIM 1将使用CRISPR-CAS9 在人类干细胞中进行编辑，然后将其分化为内皮细胞以识别因果变异以及介导4q12冠状动脉疾病GWAS基因座中关联的上游转录因子。 AIM 2将定义远端基因和受内皮静止影响的途径并研究其细胞后果是从评估休息在间充质转变中的静止作用开始。 AIM 3将使用他莫昔芬可诱导的内皮特异性休息敲除小鼠模型，以评估静止的体内效应内皮和动脉粥样硬化。该项目的成功得到了多学科心理团队的支持，包括一名血管生物学家（Charles Lowenstein博士），一名计算生物学家（Alexis Battle）和功能性遗传学专家（Andrew McCallion博士）以及血管生物学专家咨询委员会细胞分化和动脉粥样硬化（Harry Dietz博士，Chulan Kwon博士和Thomas Queermentous博士）。这奖励期将帮助PI提高其基因组技能，获得新的湿实验室技能并生成初步数据成功竞争R01资金，以将遗传见解转化为新的机制 ASCVD。