Id3 and VSMC in Murine and Human Atherosclerosis

Id3 和 VSMC 在小鼠和人类动脉粥样硬化中的作用

基本信息

批准号：
10210435
负责人：
Coleen A McNamara
金额：
$ 67.1万
依托单位：
UNIVERSITY OF VIRGINIA
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-08-28 至 2023-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10210435
关键词：
Alanine Alleles Amino Acids Animals Aorta Apolipoprotein E Arterial Fatty Streak Arteries Atherosclerosis Attenuated Automobile Driving Binding Biology Blood Vessels CRISPR/Cas technology Cause of Death Cell Adhesion Molecules Cell Count Cell Lineage Cell physiology Cells Cellular biology Characteristics Collaborations Coronary Stenosis Coronary artery Cytometry Data Deposition Development Diabetes Mellitus Disease Gene Expression Gene Mutation Genes Genetic Engineering Genetic Polymorphism Genetic Risk Genomics Growth Heart Heritability Histology Human Image Image Analysis Immune Inflammation Inflammation Mediators Inflammatory Iowa Knock-out Lead Lesion Link Measures Medial Mediating Minor Modeling Molecular Morbidity - disease rate Multi-Ethnic Study of Atherosclerosis Multivariate Analysis Mus Necrosis Null Lymphocytes Participant Phenotype Pluripotent Stem Cells Pre-Clinical Model Prevention Process Proteomics Reproducibility Research Design Residual state Reverse Transcriptase Polymerase Chain Reaction Risk Role Series Single Nucleotide Polymorphism Smooth Muscle Myocytes Stimulus System Techniques Testing Thick Threonine Translating Tumor-infiltrating immune cells Ultrasonography Universities Variant Vascular Cell Adhesion Molecule-1 Vascular Smooth Muscle calcification cell growth cohort coronary artery calcium coronary plaque dimer genome editing human subject indexing machine learning algorithm macrophage mortality mouse model new therapeutic target novel oxidized low density lipoprotein promoter risk variant transcriptome sequencing vascular smooth muscle cell proliferation virtual

项目摘要

Despite major advances in prevention and treatment of atherosclerotic cardiovascular disease (ASCVD), it remains a major cause of deaths worldwide. As ASCVD is a heritable and polygenic disorder, identification of ASCVD-associated single nucleotide polymorphisms (SNP) in humans and elucidation of their effect on ASCVD biology is needed to fully understand and impact on this unmet residual risk. One such human polymorphism in the ID3 gene (rs11574) was associated with ASCVD in 3 distinct human cohorts. Notably, this SNP significantly attenuates ID3 function and studies in pre-clinical models confirm a critical role for ID3 in atheroprotection. We have previously shown that loss of ID3 inhibits vascular smooth muscle cell (VSMC) proliferation and promotes VSMC expression of inflammatory factors, processes linked to increase atherosclerosis. In collaboration with Dr. Gary Owens, we have generated a mouse model with VSMC lineage tracing and VSMC-specific deletion of ID3 to study ID3-dependent VSMC specific changes during the course of lesion development in the context of the whole animal. We have developed a comprehensive CyTOF panel to identify and quantitate key cellular and intracellular changes in VSMC and other lesional cells in the aorta during the course of atherosclerosis development in these mice. To translate murine findings to humans, we have utilized CRISPR/Cas9 genetic engineering to knock out ID3 and produce the full allelic series of rs11574 in human cells and validated a reproducible system for differentiation of human inducible pluripotent stem cells (iPSCs) to VSMC. We will quantify the changes in VSMC phenotype in iPSC-derived VSMC with the full allelic series of rs11574. To further translate murine findings to humans, intravascular ultrasound virtual histology (IVUS-VH) will be measured in human coronary arteries and analyzed with cutting edge image analysis techniques in collaboration with Dr. Milan Sonka at the University of Iowa. As such, our group is uniquely poised to dissect the role of this ASCVD-associated SNP and translate findings to humans. We hypothesize: that loss of ID3 in VSMCs will inhibit VSMC growth and promote a phenotype that exacerbates vessel wall inflammation and atherosclerosis lesion development; that human ID3 encoded by the risk allele aggravates these atherogenic VSMC functions; and that human subjects with the risk allele have larger coronary plaques with increased fatty deposits, necrotic cores and calcification compared to subjects homozygous for the common allele.

尽管动脉粥样硬化性心血管疾病（ASCVD）的预防和治疗取得了重大进展，但仍然是全世界死亡的主要原因。由于 ASCVD 是一种遗传性多基因疾病，因此识别人类 ASCVD 相关单核苷酸多态性 (SNP) 及其对疾病影响的阐明需要 ASCVD 生物学来充分理解和影响这种未满足的残余风险。有这样一个人在 3 个不同的人类群体中，ID3 基因 (rs11574) 的多态性与 ASCVD 相关。尤其，该 SNP 显着减弱 ID3 功能，临床前模型研究证实 ID3 在动脉粥样硬化保护。我们之前已经证明 ID3 的缺失会抑制血管平滑肌细胞 (VSMC) 增殖并促进 VSMC 表达炎症因子，与增加相关的过程动脉粥样硬化。我们与 Gary Owens 博士合作，生成了具有 VSMC 谱系的小鼠模型跟踪和 VSMC 特异性 ID3 删除，以研究课程期间 ID3 依赖性 VSMC 特异性变化在整个动物的背景下病变的发展。我们开发了全面的 CyTOF 面板识别和定量 VSMC 和主动脉其他病变细胞的关键细胞和细胞内变化在这些小鼠的动脉粥样硬化发展过程中。为了将小鼠的发现转化为人类，我们利用 CRISPR/Cas9 基因工程敲除 ID3 并产生 rs11574 的完整等位基因系列在人类细胞中，并验证了用于人类诱导多能干细胞分化的可重复系统 (iPSC) 至 VSMC。我们将使用完整的等位基因来量化 iPSC 衍生的 VSMC 中 VSMC 表型的变化 rs11574系列。为了进一步将小鼠发现转化为人类，血管内超声虚拟组织学 (IVUS-VH) 将在人体冠状动脉中进行测量，并通过尖端图像分析进行分析与爱荷华大学的 Milan Sonka 博士合作的技术。因此，我们的团队是独一无二的准备剖析这种与 ASCVD 相关的 SNP 的作用并将研究结果转化为人类。我们假设： VSMC 中 ID3 的缺失将抑制 VSMC 生长并促进加剧血管恶化的表型管壁炎症和动脉粥样硬化病变发展；由风险等位基因编码的人类 ID3 加剧这些致动脉粥样硬化的 VSMC 功能；具有风险等位基因的人类受试者有更大的与受试者相比，冠状动脉斑块脂肪沉积、坏死核心和钙化增加共同等位基因纯合。