The SMAD3 signaling network in coronary artery disease risk

SMAD3 信号网络在冠状动脉疾病风险中的作用

• 批准号: 10077579
• 负责人: THOMAS QUERTERMOUS
• 金额: $ 57.6万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-01-15 至 2022-10-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10077579
• 关键词: 15q22; 1p36; Address; Affect; Alleles; Anatomy; Apolipoprotein E; Atherosclerosis; Binding; Bioinformatics; Blood Vessels; Cardiovascular Diseases; Cell Lineage; Cell Proliferation; Cell physiology; Cells; Cellular biology; ChIP-seq; Coronary; Coronary Arteriosclerosis; Coronary artery; Data; Data Set; Development; Disease; Enhancers; Etiology; Gene Expression; Gene Expression Profiling; Gene Family; Genes; Genetic; Genetic Models; Genetic Risk; Genetic Transcription; Genomic approach; Haplotypes; Human; Human Genome; In Vitro; Introns; Laboratories; Learning; Lesion; Link; MADH3 gene; Medial; Mediating; Modeling; Molecular; Mus; Pathway interactions; Pattern; Phenotype; Phosphotransferases; Process; Quantitative Trait Loci; Reporter; Research; Risk; Risk Assessment; Role; Rupture; SKI gene; Scientist; Signal Pathway; Signal Transduction; Signaling Molecule; Smooth Muscle Myocytes; Stimulus; Structure; Syndrome; Therapeutic; Tissues; Transforming Growth Factor beta; Transgenic Organisms; Variant; Vascular Diseases; Work; causal variant; cell dedifferentiation; cell type; disorder risk; epigenome; experimental study; genetic approach; genome editing; genome wide association study; genomic locus; histone modification; in vivo; innovation; interest; macrophage; molecular phenotype; mouse genetics; mouse model; programs; protective allele; public health relevance; receptor; response; single-cell RNA sequencing; therapeutic development; transcription factor; transcriptome; transcriptome sequencing; vascular stress; 15q22; 1p36; Address; Affect; Alleles; Anatomy; Apolipoprotein E; Atherosclerosis; Binding; Bioinformatics; Blood Vessels; Cardiovascular Diseases; Cell Lineage; Cell Proliferation; Cell physiology; Cells; Cellular biology; ChIP-seq; Coronary; Coronary Arteriosclerosis; Coronary artery; Data; Data Set; Development; Disease; Enhancers; Etiology; Gene Expression; Gene Expression Profiling; Gene Family; Genes; Genetic; Genetic Models; Genetic Risk; Genetic Transcription; Genomic approach; Haplotypes; Human; Human Genome; In Vitro; Introns; Laboratories; Learning; Lesion; Link; MADH3 gene; Medial; Mediating; Modeling; Molecular; Mus; Pathway interactions; Pattern; Phenotype; Phosphotransferases; Process; Quantitative Trait Loci; Reporter; Research; Risk; Risk Assessment; Role; Rupture; SKI gene; Scientist; Signal Pathway; Signal Transduction; Signaling Molecule; Smooth Muscle Myocytes; Stimulus; Structure; Syndrome; Therapeutic; Tissues; Transforming Growth Factor beta; Transgenic Organisms; Variant; Vascular Diseases; Work; causal variant; cell dedifferentiation; cell type; disorder risk; epigenome; experimental study; genetic approach; genome editing; genome wide association study; genomic locus; histone modification; in vivo; innovation; interest; macrophage; molecular phenotype; mouse genetics; mouse model; programs; protective allele; public health relevance; receptor; response; single-cell RNA sequencing; therapeutic development; transcription factor; transcriptome; transcriptome sequencing; vascular stress

PROJECT SUMMARY/ABSTRACT The TGFβ signaling pathway has been extensively studied in vascular disease, but there remains considerable controversy regarding the direction and mechanism of effect for how this pathway impacts human coronary artery disease (CAD). Recent genome-wide association studies have identified several loci that harbor TGFβ family genes, including the SMAD3 gene at 15q22.33 that encodes a transcription factor critical for converting TGFβ-induced cytoplasmic signaling to gene expression changes, and ZEB2 and SKI at 2q22.3 and 1p36.33 respectively, which bind SMAD3 and modulate its transcriptional activity. Studies in this lab have employed histone modification, chromosomal accessibility, allele-specific expression, in vitro genome editing and transgenic reporter mouse studies to identify SMAD3 as the causal gene at 15q22.33. The protective allele for this gene disrupts a potent intronic enhancer in the SMAD3 gene that is associated with decreased SMAD3 expression in vascular tissues, suggesting that expression of SMAD3 in SMC promotes risk for CAD. In SMC, TGFβ signaling is known to have important differentiative and anti-proliferative roles during vascular development, but this function may be deleterious in the disease setting where SMC dedifferentiation and proliferation, “phenotypic modulation,” allows this cell type to bolster structural integrity and retard plaque rupture. A disease-promoting role for TGFβ is supported by our studies with TCF21, a CAD associated transcription factor that promotes SMC phenotypic modulation and whose protective allele confers increased expression. Taken together, these data suggest our Central Hypothesis: the TGFβ signaling molecule SMAD3, in conjunction with ZEB2 and SKI, regulates a transcriptional network that mediates the adaptive SMC phenotypic response to vascular stress, with allelic variation modulating this response contributing to CAD risk. Experiments proposed in Aim 1 in the ApoE-/- atherosclerosis mouse model will evaluate disease anatomy with SMC-specific deletion of Smad3, along with human risk and protective haplotypes created by genome editing. Lineage tracing and single cell RNA-seq will define the role of Smad3 in regulating the cellular response to disease stimuli, and molecular phenotype as lesion SMC dedifferentiate and contribute to the macrophage lineage. In Aim 2, ChIP-seq and RNA-seq studies in human coronary artery SMC will define the network of genes that are regulated by SMAD3 and investigate how expression of ZEB2 and SKI modulates the molecular composition of this network. In vitro studies in human coronary artery SMC in Aim 3 will identify the cellular and molecular processes that are mediated by SMAD3 and how these functions are modified by ZEB2 and SKI. Taken together, these studies will significantly advance our understanding of how SMAD3 and related factors ZEB2 and SKI govern the SMC phenotypic response to vascular disease, and how perturbation of their function contributes to CAD risk.

项目摘要/摘要 TGFβ信号通路已在血管疾病中广泛研究，但仍存在 关于该途径如何影响的效果方向和机理的争议很大 人类冠状动脉疾病（CAD）。最近的全基因组关联研究已经确定了几个 携带TGFβ家族基因的基因座，包括编码转录的15q22.33的Smad3基因 将TGFβ诱导的细胞质信号转化为基因表达变化的关键因素，而Zeb2 分别在2q22.3和136.33处滑雪，它们结合Smad3并调节其转录活性。 该实验室的研究已经进行了组蛋白的修饰，染色体可及性，等位基因特异性 表达，体外基因组编辑和转基因报告基因小鼠研究，以识别SMAD3为因果关系 基因在15q22.33。该基因的受保护等位基因破坏了SMAD3中潜在的内含子增强子 与血管组织中Smad3表达降低相关的基因，表明表达 SMC中的SMAD3促进了CAD的风险。在SMC中，已知TGFβ信号具有重要 血管发育过程中的区分和反增殖作用，但可以删除此功能 在SMC去分化和增殖的疾病环境中，“表型调节”允许这一点 细胞类型至增强结构完整性和延迟斑块破裂。 TGFβ的促进疾病的作用 在我们对TCF21的研究（促进SMC表型的CAD相关转录因子）的研究中支持 调节且其受保护的等位基因承认增加了表达。综上所述，这些数据表明 我们的中心假设：TGFβ信号分子SMAD3与Zeb2和Ski结合使用， 调节转录网络，该网络介导适应性SMC表型反应 血管胁迫，倾斜变化调节这种反应导致CAD风险。 APOE - / - 动脉粥样硬化小鼠模型中AIM 1提出的实验将评估疾病解剖结构 SMAD3的SMC特异性缺失，以及人类风险和受基因组创建的受保护的单倍型 编辑。谱系跟踪和单细胞RNA-seq将定义SMAD3在调节细胞中的作用 对疾病刺激的反应和分子表型作为病变SMC去分化，并有助于 巨噬细胞谱系。在AIM 2中，人类冠状动脉中的CHIP-SEQ和RNA-SEQ研究将定义 由Smad3调节并研究Zeb2和Ski的表达方式的基因网络 调节该网络的分子组成。 AIM中人类冠状动脉动脉的体外研究 3将确定由Smad3介导的细胞和分子过程以及这些功能如何 由Zeb2和滑雪修改。综上所述，这些研究将大大提高我们的理解 SMAD3和相关因素Zeb2和Ski如何控制SMC表型对血管的反应 疾病，以及其功能的扰动如何导致CAD风险。