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

• 批准号: 10004164
• 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
• 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/10004164
• 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

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，以研究课程期间依赖 ID3 的 VSMC 特异性变化 在整个动物的背景下损伤的发展。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。