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

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.

项目总结/摘要 TGFβ信号通路在血管疾病中已被广泛研究，但仍存在 关于该途径如何影响的方向和作用机制存在相当大的争议 人冠状动脉疾病（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，包括位于15q22.33的SMAD 3基因，该基因编码转录 转化生长因子β诱导的细胞质信号传导转化为基因表达变化的关键因子，以及ZEB 2 和SKI分别位于2q22.3和1p36.33，它们结合SMAD 3并调节其转录活性。 该实验室的研究采用了组蛋白修饰，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小鼠研究，以确定SMAD 3为致病基因。 基因位于15q22.33。该基因的保护性等位基因破坏了SMAD 3中一个有效的内含子增强子。 一个与SMAD 3在血管组织中表达降低相关的基因，表明SMAD 3的表达 SMC中SMAD 3的表达增加了CAD的风险。在SMC中，已知TGFβ信号传导具有重要的 分化和抗增殖的作用，但这种功能可能是有害的 在疾病背景下，SMC去分化和增殖，“表型调节”， 细胞类型，以支持结构完整性和延缓斑块破裂。TGFβ的一种疾病促进作用是 我们对TCF 21的研究支持了这一点，TCF 21是一种CAD相关的转录因子，可促进SMC表型 调节并且其保护性等位基因赋予增加的表达。综合来看，这些数据表明 我们的中心假设：TGFβ信号分子SMAD 3，与ZEB 2和SKI结合， 调节介导适应性SMC表型反应的转录网络， 血管应激，等位基因变异调节这种反应，有助于CAD的风险。 目标1中提出的在ApoE-/-动脉粥样硬化小鼠模型中的实验将评估疾病解剖学 SMC特异性Smad 3缺失，沿着人类风险和保护性单倍型， 编辑.谱系追踪和单细胞RNA-seq将确定Smad 3在调节细胞凋亡中的作用。 对疾病刺激的反应，以及病变SMC去分化的分子表型，并有助于 巨噬细胞系在目标2中，人冠状动脉SMC中的ChIP-seq和RNA-seq研究将定义 SMAD 3调控的基因网络，并研究ZEB 2和SKI的表达 调节了这个网络的分子组成。Aim中人冠状动脉SMC的体外研究 3将确定SMAD 3介导的细胞和分子过程以及这些过程如何发挥作用 由ZEB 2和SKI修饰。综合起来，这些研究将大大推进我们的理解 SMAD 3和相关因子ZEB 2和SKI如何控制SMC对血管内皮细胞的表型反应， 疾病，以及它们的功能扰动如何导致CAD风险。

项目成果

Advances in Transcriptomics: Investigating Cardiovascular Disease at Unprecedented Resolution.

• DOI: 10.1161/circresaha.117.310910
• 发表时间: 2018-04-27
• 期刊: Circulation research
• 影响因子: 20.1
• 作者: Wirka RC;Pjanic M;Quertermous T
• 通讯作者: Quertermous T

Single-nucleus chromatin accessibility profiling highlights regulatory mechanisms of coronary artery disease risk.

• DOI: 10.1038/s41588-022-01069-0
• 发表时间: 2022-06
• 期刊: Nature genetics
• 影响因子: 30.8
• 作者: Turner AW;Hu SS;Mosquera JV;Ma WF;Hodonsky CJ;Wong D;Auguste G;Song Y;Sol-Church K;Farber E;Kundu S;Kundaje A;Lopez NG;Ma L;Ghosh SKB;Onengut-Gumuscu S;Ashley EA;Quertermous T;Finn AV;Leeper NJ;Kovacic JC;Björkegren JLM;Zang C;Miller CL
• 通讯作者: Miller CL

Coronary artery disease genes SMAD3 and TCF21 promote opposing interactive genetic programs that regulate smooth muscle cell differentiation and disease risk.

• DOI: 10.1371/journal.pgen.1007681
• 发表时间: 2018-10
• 期刊: PLoS genetics
• 影响因子: 4.5
• 作者: Iyer D;Zhao Q;Wirka R;Naravane A;Nguyen T;Liu B;Nagao M;Cheng P;Miller CL;Kim JB;Pjanic M;Quertermous T
• 通讯作者: Quertermous T