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.

尽管动脉粥样硬化心血管疾病（ASCVD）的预防和治疗方面取得了重大进展，但 仍然是全球死亡的主要原因。由于ASCVD是一种可遗传的多基因障碍，因此 ASCVD相关的单核苷酸多态性（SNP）在人类中及其对其对影响的影响 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 ID3基因（RS11574）中的多态性与3个不同的人类人群中的ASCVD相关。尤其， 该SNP显着削弱了ID3功能，并且在临床前模型中的研究证实了ID3的关键作用 动脉保护。我们以前已经表明，ID3的损失抑制了血管平滑肌细胞（VSMC） 增殖并促进炎症因子的VSMC表达，与增加有关的过程 动脉粥样硬化。与Gary Owens博士合作，我们生成了一个使用VSMC Lineage的鼠标模型 ID3的跟踪和VSMC特异性删除在课程过程中研究ID3依赖性VSMC特定变化 在整个动物的背景下的病变发展。我们已经开发了一个全面的细胞面板 识别和定量主动脉中VSMC和其他病变细胞中的钥匙细胞和细胞内变化 在这些小鼠的动脉粥样硬化发展过程中。为了将鼠的发现转化为人类，我们 已经利用CRISPR/CAS9基因工程来淘汰ID3并制作完整的等位基因系列RS11574 在人类细胞中，并验证了可再现的系统，用于分化人诱导的多能干细胞 （IPSC）到VSMC。我们将量化具有完整等位基因的IPSC衍生VSMC中VSMC表型的变化 系列RS11574。为了进一步将鼠的发现转化为人类，血管内超声虚拟组织学 （IVUS-VH）将在人类冠状动脉中进行测量，并通过尖端图像分析进行分析 与爱荷华大学米兰·桑卡（Milan Sonka）博士合作的技术。因此，我们的小组是独特的 准备剖析与ASCVD相关的SNP的作用，并将发现转化为人类。我们假设： VSMC中ID3的损失将抑制VSMC的增长并促进加剧血管的表型 壁炎和动脉粥样硬化病变的发育；那个由风险等位基因编码的人ID3 加剧了这些动脉粥样硬化的VSMC功能；并且具有风险等位基因的人类受试者的受试者有更大的 与受试者相比 共同等位基因的纯合子。