TCF21 is a causal coronary artery disease gene that modulates coronary smooth muscle phenotypic transition via epigenetic mechanisms

TCF21 是一种致病性冠状动脉疾病基因，可通过表观遗传机制调节冠状动脉平滑肌表型转变

• 批准号: 10536419
• 负责人: DANIEL YUHANG LI
• 金额: $ 6.68万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-02 至 2025-09-01
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10536419
• 关键词: ATAC-seq; Address; Anatomy; Animals; Apolipoprotein E; Arterial Fatty Streak; Atherosclerosis; BHLH Protein; Binding; Biological Assay; Blood Vessels; Cardiometabolic Disease; Cardiovascular Pathology; Cell Lineage; Cells; Characteristics; Chromatin; Clinical; Computational Technique; Coronary; Coronary Arteriosclerosis; Coronary heart disease; Development; Disease; Epigenetic Process; Evaluation; Fibroblasts; Functional disorder; Gene Expression; Genes; Genetic; Genetic Transcription; Genetic study; Goals; High Fat Diet; Knock-out; Laboratories; Lesion; Lipids; Maps; Mediating; Mentors; Modeling; Molecular; Mus; Phenotype; Physicians; Regulator Genes; Regulatory Pathway; Research; Research Personnel; Scientist; Smooth Muscle; Smooth Muscle Myocytes; Stress; System; Therapeutic; Time; Tissues; Transgenic Organisms; Transitional Cell; Translating; Variant; Vascular Diseases; Vascular Smooth Muscle; Work; disorder risk; epigenomics; gene network; gene regulatory network; genome wide association study; genome-wide; genomic locus; in vivo; migration; mouse model; multiple omics; novel; overexpression; phenotypic biomarker; prevent; protective effect; response; risk variant; single cell analysis; single-cell RNA sequencing; therapeutic target; tool; transcription factor; transcriptome sequencing; transcriptomics; vascular stress

PROJECT SUMMARY More than 161 genetic loci have been associated with coronary artery disease (CAD) through the genome wide association studies (GWAS) conducted by the CARDIoGRAMplusC4D consortium. However, despite such advances in the understanding of coronary disease, therapeutic targets beyond lipid lowering therapies have yet to reach the clinical stage. At this critical juncture for discovery, there is a need for better mechanistic characterization of the genetic CAD risk loci at the single cell level to simultaneously study the genetic and epigenetic phenomena underlying disease risk. The Quertermous laboratory has previously identified TCF21 as the CAD associated gene mapped by GWAS at the 6q23.2 locus. A combination of smooth muscle cell (SMC) lineage tracing and single cell RNA sequencing has shown that Tcf21 is upregulated and promotes SMC de-differentiation, proliferation and migration into atherosclerotic plaque. TCF21 expression contributes to the phenotypic transition of SMC to the protective fibrous cap, producing fibroblast-like cells termed “fibromyocytes” (FMC). We are now studying this transitional state at multiple developmental time points with single cell transcriptomics and epigenomics in the ApoE knockout model of atherosclerosis and find wide variation in chromatin accessibility associated with the SMC-FMC transitional cell state. While loss of Tcf21 produces more than a 50% decrease in FMC, there remains a subpopulation of SMC that can contribute to FMC and the fibrous cap. The gene regulatory networks that cooperate with TCF21 to promote SMC transition, and the downstream target genes of these networks remain unknown. The central hypothesis underlying this proposal is that TCF21 exerts a protective effect toward CAD risk through its ability to promote phenotypic transition of SMC in the setting of vascular wall stress. To address this hypothesis, the applicant will study a SMC-specific model of Tcf21 over-expression in the setting of vascular disease and characterize the disease consequences of increased Tcf21 action through cutting-edge transcriptomic and epigenetic tools to simultaneously evaluate single cell gene transcription and chromatin accessibility through combined scRNAseq with scATACseq in vascular tissue. Aim 1 will examine the overall effect of increased Tcf21 expression on SMC phenotype as well as atherosclerotic plaque cellular composition and anatomy in transgenic over-expressing animals. Aim 2 will elucidate the transcriptional and epigenetic mechanisms by which Tcf21 mediates SMC de-differentiation and transition to the FMC phenotype using analysis of single cell multi-omic RNAseq and ATACseq. This study will address fundamental questions related to vascular disease pathophysiology, including: i) does TCF21 over-expression increase the SMC to FMC transition, ii) does increased SMC to FMC transition promote a more stable plaque phenotype, and iii) how does TCF21 regulate the epigenetic and transcriptional landscape to promote SMC transition to a protective FMC phenotype.

项目摘要 超过161个遗传基因座通过基因组与冠状动脉疾病（CAD）有关 Cardiogramplusc4d联盟进行的广泛关联研究（GWAS）。但是，需求 在理解冠状动脉疾病的情况下，脂质降低疗法以外的治疗靶点 尚未达到临床阶段。在这个关键的发现时，需要更好的机理 遗传CAD CAD风险基因座在单细胞水平上的表征同时研究遗传和 疾病风险的表观遗传现象。查询实验室先前已经确定了TCF21 作为CAD相关的基因由GWA映射在6q23.2基因座。平滑肌细胞的组合 （SMC）谱系跟踪和单细胞RNA测序表明TCF21已更新并促进 SMC去分化，增殖和迁移到动脉粥样硬化斑块中。 TCF21表达有助于 SMC向受保护的纤维帽的表型过渡，产生称为成纤维细胞样细胞 “纤维细胞”（FMC）。我们现在正在以多个发展时间点研究这种过渡状态 动脉粥样硬化的APOE基因敲除模型中的单细胞转录组学和表观基因组学，发现广泛 与SMC-FMC过渡细胞状态相关的染色质可及性的变化。而TCF21的损失 FMC产生超过50％的降低，SMC的亚群可以有助于 FMC和纤维帽。与TCF21合作以促进SMC过渡的基因调节网络， 这些网络的下游靶基因仍然未知。中央假设的基础 该建议是TCF21通过其促进能力对CAD风险执行受保护的效果 SMC在血管壁应力的环境中的表型过渡。为了解决这一假设， 申请人将在血管疾病和 通过尖端的转录组和 同时评估单细胞基因转录和染色质访问性的表观遗传学工具 在血管组织中与scatacseq合并。 AIM 1将检查增加的总体效果 TCF21在SMC表型以及动脉粥样硬化斑块细胞组成和解剖学上的表达 转基因过表达动物。 AIM 2将通过 TCF21通过分析单细胞介导SMC脱离分化和过渡到FMC表型 多摩变RNASEQ和ATACSEQ。这项研究将解决与血管疾病有关的基本问题 病理生理学，包括：i）dis tcf21过表达增加了SMC到FMC转变，ii） 将SMC增加到FMC过渡会促进更稳定的斑块表型，iii）TCF21如何调节 表观遗传和转录景观，以促进SMC过渡到受保护的FMC表型。