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，但它 仍然是世界范围内死亡的主要原因。由于ASCVD是一种可遗传的多基因疾病，因此确定 人类ASCVD相关单核苷酸多态(SNP)及其对ASCVD的影响 需要ASCVD生物学来充分了解这一未得到满足的残余风险并对其产生影响。一个这样的人 在3个不同的人群中，ID3基因(Rs11574)多态性与ASCVD相关。值得注意的是， 这种SNP显著减弱ID3的功能，临床前模型的研究证实ID3在 动脉粥样硬化保护。我们先前已经证明ID3的丢失抑制血管平滑肌细胞(VSMC) 促增殖和促进VSMC表达炎症因子，过程中相关增加 动脉硬化。在与加里·欧文斯博士的合作下，我们创造了一个具有VSMC血统的小鼠模型 追踪和VSMC特异性删除ID3以研究ID3依赖的VSMC在过程中的特异性变化 在整个动物的背景下病变的发展。我们已经开发了一种全面的CyTOF面板 识别和量化主动脉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。为了将老鼠的发现转化为人类，我们 利用CRISPR/Cas9基因工程敲除id3，产生rs11574的全等位基因序列 并验证了人可诱导多能干细胞分化的可重复性系统 (IPSCS)到VSMC。我们将用全等位基因来量化IPSC来源的VSMC表型的变化 系列rs11574。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)将在人的冠状动脉中测量并用尖端图像分析进行分析 技术与爱荷华大学的米兰·松卡博士合作。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，并将发现转化为人类。我们假设： VSMC中ID3的丢失将抑制VSMC的生长，并促进血管恶化的表型 壁炎症和动脉粥样硬化病变发展；人类ID3由危险等位基因编码 加重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，脂肪沉积、坏死核和钙化增加的冠状动脉斑块 常见等位基因的纯合子。