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: 10421070
• 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/10421070
• 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; 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; 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; induced pluripotent stem cell; machine learning algorithm; macrophage; mortality; mouse model; new therapeutic target; novel; oxidized low density lipoprotein; promoter; risk variant; transcriptome sequencing; ultrasound; 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.

尽管在预防和治疗动脉粥样硬化性心血管疾病（ASCVD）方面取得了重大进展， 仍然是全球死亡的主要原因。由于ASCVD是一种遗传性多基因疾病， 人类ASCVD相关的单核苷酸多态性（SNP）及其对ASCVD的影响 需要ASCVD生物学来充分理解和影响这种未满足的剩余风险。一个这样的人类 在3个不同的人群中，ID 3基因（rs 11574）多态性与ASCVD相关。值得注意地是， 该SNP显著减弱ID 3功能，并且在临床前模型中的研究证实了ID 3在以下方面的关键作用： 动脉粥样硬化保护我们先前已经表明，ID 3的缺失抑制血管平滑肌细胞（VSMC） 增殖和促进VSMC表达炎症因子，过程联系到增加 动脉粥样硬化与加里欧文斯博士合作，我们已经产生了一个小鼠模型与VSMC谱系 追踪和VSMC特异性ID 3缺失，以研究病程中ID 3依赖性VSMC特异性变化 在整个动物的背景下，病变发展。我们开发了一个全面的CyTOF面板 识别和定量主动脉中VSMC和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ID 3并产生rs 11574的全等位基因系列 并验证了用于人诱导性多能干细胞分化的可再现系统 （iPSC）到VSMC。我们将量化具有完整等位基因的iPSC衍生的VSMC中VSMC表型的变化， RS 11574系列。为了进一步将小鼠的发现转化为人类，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，并使用尖端图像分析进行分析 这项技术是与爱荷华州大学的米兰·桑卡博士合作完成的。因此，我们的团队是独一无二的 准备剖析这种ASCVD相关SNP的作用，并将发现转化为人类。我们假设： VSMC中ID 3的缺失将抑制VSMC的生长并促进一种表型， 壁炎症和动脉粥样硬化病变发展;由风险等位基因编码人ID 3 增强了这些致动脉粥样硬化的VSMC功能;并且具有风险等位基因的人类受试者具有更大的 与受试者相比，脂肪沉积、坏死核心和钙化增加的冠状动脉斑块 是普通等位基因的纯合子