PDGFD regulates a transcriptional network to modulate smooth muscle cell transition and coronary artery disease risk

PDGFD 调节转录网络以调节平滑肌细胞转变和冠状动脉疾病风险

基本信息

批准号：
10593934
负责人：
THOMAS QUERTERMOUS
金额：
$ 67.51万
依托单位：
STANFORD UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-04-15 至 2025-03-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10593934
关键词：
AHR gene ATAC-seq Alleles Anatomy Animals Apolipoprotein E Aryl Hydrocarbon Receptor Atherosclerosis Autocrine Communication Binding Blood Vessels CRISPR interference Cardiovascular Diseases Cell Lineage Cell model Cells Chromatin Chromosome Mapping Complement Factor D Coronary Arteriosclerosis Coronary artery Coupled Data Development Disease Enhancers Epigenetic Process Fibroblasts Functional disorder Gene Expression Gene Targeting Genes Genetic Risk Genetic Transcription Goals Growth Factor Human Human Genome In Vitro Knock-out Knockout Mice LacZ Genes Link MADH3 gene Maps Measures Mediating Modeling Molecular Morphology Mus PDGFA gene PDGFRB gene Phenotype Platelet-Derived Growth Factor Probability Process Regulation Reporter Research Risk Assessment Role Scientist Signal Pathway Signal Transduction Smooth Muscle Myocytes Stromal Cell-Derived Factor 1 Subcellular Anatomy TWIST1 gene Therapeutic Tissues Transcription Process Transcriptional Activation Wild Type Mouse Work cell dedifferentiation cell type disorder risk gene function genome wide association study genome-wide in vivo mouse model programs public health relevance receptor response single-cell RNA sequencing transcription factor transcriptomic profiling transcriptomics vascular stress whole genome

项目摘要

We have identified TCF21 as the coronary artery disease (CAD) associated gene mapped by genome-wide association studies at 6q23.2 and employed numerous mechanistic approaches to show that it promotes a smooth muscle cell (SMC) transition to a fibroblast like “fibromyocyte” phenotype, and the contribution of these cells to the protective fibrous cap. Our studies with another CAD associated gene, the aryl hydrocarbon receptor (AHR), have characterized the transition of SMC to a second, chondrogenic “chondromyocyte” phenotype. To extend this work and investigate the mechanisms of epigenetic signaling upstream of TCF21, AHR, and other factors that mediate SMC cell state, we are focusing efforts on the CAD associated platelet derived growth factor D gene (PDGFD). We have shown that PDGFD regulates TCF21 and other validated CAD genes including LMOD1, CXCL12, and SMAD3, and is expressed primarily in disease transition SMC that also express the PDGFRB receptor. Together, these data suggest that PDGFD activates an autocrine signaling pathway that modulates SMC phenotype and CAD risk. The hypothesis directing this research postulates that PDGFD promotes CAD risk through its regulation of TCF21 and other key disease related transcription factors that mediate the SMC phenotypic response to vascular stress. The primary goals of the work proposed here are thus to identify the PDGFD target transcription factors (TFs) that regulate SMC transitions and characterize their transcriptional program in this cell type. Specifically, in Aim 1 we will employ Pdgfd knockout and SMC lineage tracing in the ApoE null mouse atherosclerosis model to characterize the effect of this gene on SMC cell state transitions, and the impact of perturbing these transitions on disease morphology and cellular anatomy. In Aim 2, we will conduct single cell RNA sequencing (scRNAseq) in Pdgfd null and wildtype atherosclerotic mice to characterize the SMC gene expression program downstream of Pdgfd in this cell type. Single cell ATAC sequencing (scATACseq) in the same animals will map enhancers genome- wide that are differentially regulated in SMC phenotypic transitions, and identify specific TFs that bind these enhancers to regulate expression of fibromyocyte and chondromyocyte specific genes. In Aim 3, we will perturb candidate SMC transition promoting TFs that are identified in Aim 2, in vitro in a PDGFD stimulated human coronary artery smooth muscle cell de-differentiation model, and the resulting transcriptomic and cell state effects interpreted in the context of PDGFD function in this model. These studies will link PDGFD to CAD associated genes that we have characterized in the context of SMC phenotypic transition (TCF21, AHR, SMAD3, TWIST1), and to additional high probability CAD genes that regulate SMC phenotype, to expand the disease transcriptional network in this vascular cell type. This work will advance our understanding of atherosclerosis pathophysiology and promote efforts to target vascular wall molecular processes to ameliorate CAD risk.

我们已经确定TCF21是由基因组绘制的冠状动脉疾病（CAD）在6q23.2的协会研究，并采用了多种机械方法来表明它促进了平滑肌细胞（SMC）过渡到像“纤维细胞”表型这样的成纤维细胞，并且这些贡献细胞到受保护的纤维帽。我们使用另一个CAD相关基因的研究芳基烃受体（AHR）已经表征了SMC向第二个软骨的“软骨细胞”的过渡表型。为了扩展这项工作并研究TCF21上游表观遗传信号的机制， AHR和其他介导SMC细胞状态的因素，我们将精力集中在CAD相关的血小板上衍生生长因子D基因（PDGFD）。我们已经表明，PDGFD调节TCF21和其他经过验证包括LMOD1，CXCL12和SMAD3在内的CAD基因，主要在疾病过渡中表达还表达PDGFRB受体。这些数据一起表明PDGFD激活自分泌调节SMC表型和CAD风险的信号通路。指导这项研究的假设假设PDGFD通过调节TCF21和其他关键疾病来促进CAD风险介导SMC表型反应对血管应激的相关转录因子。主要因此，这里提出的工作的目标是确定调节的PDGFD目标转录因子（TF） SMC过渡并在此单元格中表征其转录程序。具体来说，在AIM 1中，我们将员工PDGFD淘汰和SMC谱系跟踪在ApoE Null小鼠动脉粥样硬化模型中以表征该基因对SMC细胞状态过渡的影响以及这些过渡对疾病的影响形态和细胞解剖结构。在AIM 2中，我们将在PDGFD中进行单细胞RNA测序（SCRNASEQ） NULL和WILDTYPE动脉粥样硬化小鼠以表征PDGFD下游的SMC基因表达程序在此单元格中。同一动物中的单细胞ATAC测序（scatacseq）将绘制增强子基因组 - 在SMC表型过渡中受到不同调节的宽宽，并确定结合这些结合的特定TF 增强子来调节纤维肌细胞和软骨细胞特异性基因的表达。在AIM 3中，我们将在PDGFD刺激的PDGFD中，促进AIM 2中鉴定出在AIM 2中鉴定的TF的待遇SMC转变人类冠状动脉平滑肌细胞脱不同模型，以及所得的转录组和细胞在此模型中，在PDGFD函数的背景下解释的状态效应。这些研究将把PDGFD与CAD联系起来我们在SMC表型转变背景下表征的相关基因（TCF21，AHR， SMAD3，Twist1），以及调节SMC表型的其他高概率CAD基因，以扩展这种血管细胞类型中的疾病转录网络。这项工作将提高我们对动脉粥样硬化病理生理学并促进靶向血管壁分子过程以改善的努力 CAD风险。