Role of Smooth Muscle-derived Vascular Progenitor Cells (AdvSca1-SM) in Vasa Vasorum Expansion and Atherosclerotic Plaque Progression

平滑肌源性血管祖细胞 (AdvSca1-SM) 在血管滋养管扩张和动脉粥样硬化斑块进展中的作用

• 批准号: 10686163
• 负责人: Allison M Dubner
• 金额: $ 3.57万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-01 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10686163
• 关键词: Ablation; Acute; Arterial Fatty Streak; Atherosclerosis; Automobile Driving; Biological Assay; Blood Vessels; Cell Differentiation process; Cells; Cholesterol; Chronic; Chronic Disease; Clinical; Complex; Coronary artery; Development; Diet; Disease; Disease Progression; Endothelial Cells; Endothelium; Erinaceidae; Fatty acid glycerol esters; Fibrosis; Flow Cytometry; Gene Expression; Genes; Genetic; Heart Diseases; Hematological Disease; Histologic; Histology; Human; Immune; Immunofluorescence Microscopy; In Situ; Infiltration; Inflammatory; Injury; Knock-in; Knock-out; Knowledge; Lipids; LoxP-flanked allele; Lung diseases; Maintenance; Methods; Microscopy; Modeling; Modification; Morbidity - disease rate; Morphology; Mus; Myofibroblast; National Heart, Lung, and Blood Institute; Pathogenesis; Pathway interactions; Play; Population Sizes; Process; Public Health; Research; Role; Severities; Signal Transduction; Sleep Disorders; Smooth Muscle; Smooth Muscle Myocytes; Tamoxifen; Testing; Tunica Adventitia; Vascular Smooth Muscle; Work; beta catenin; defined contribution; diphtheria toxin receptor; falls; matrigel; meetings; mortality; mouse model; mutant; neovascularization; next generation; novel; overexpression; progenitor; recruit; single-cell RNA sequencing; stem; stem cells; vasa vasorum; vascular injury

PROJECT SUMMARY/ABSTRACT Atherosclerosis is a complex inflammatory disease and a major cause of morbidity and mortality worldwide, but current therapies fail to adequately meet clinical needs. Until recently, most atherosclerosis research has focused on endothelial activation, immune cell recruitment, and lipid retention within the intima, but emerging evidence implicates the vessel adventitia in the pathogenesis of atherosclerosis. Specifically, it has been suggested that expansion of adventitial microvessels, the vasa vasorum (VV), drives atherosclerosis progression by facilitating inflammatory cell infiltration to the vessel wall and developing atheroma. This project focuses on the role that smooth muscle-derived vascular progenitor cells, termed AdvSca1-SM cells, play in vasa vasorum expansion and plaque neovascularization. Our lab previously identified AdvSca1-SM cells as a subset of resident vascular progenitor cells reprogrammed from mature smooth muscle cells in situ in a Klf4 depended process. These cells are enriched for genes in the hedgehog/Wnt/β-catenin pathways, particularly Gli1, and have multilineage differentiation potential. In the setting of acute vascular injury, AdvSca1-SM cells expand robustly to contribute to fibrotic remodeling, but it is unclear what role they play in chronic injuries such as atherosclerosis. Work from several groups has suggested that adventitial progenitor cells can contribute to VV expansion, and our group has previously demonstrated that AdvSca1-SM cells are capable of de novo vessel formation via endothelial or smooth muscle cell differentiation in Matrigel plug assays. Additionally, we have identified AdvSca1-SM-like cells in the microvessels surrounding human coronary arteries, suggesting a potential role in the VV. The aim of this project is to determine the contribution of AdvSca1-SM cells to VV expansion, plaque neovascularization, and atherosclerotic progression. We hypothesize that AdvSca1-SM cells are major contributors to atherosclerosis, and that modulation of these cells could alter disease progression. Specific Aim 1 will define the functional contribution of AdvSca1-SM cells to VV expansion and/or plaque progression using a highly specific AdvSca1- SM genetic lineage tracing mouse model; AdvSca1-SM contribution will be assessed using immunofluorescent microscopy and flow cytometry. Specific Aim 2 will define the mechanisms controlling AdvSca1-SM cells in the setting of atherosclerosis, with an emphasis on the Gli/Wnt/β-catenin/Klf4 signaling axis previously identified as a regulator of AdvSca1-SM cells in the setting of acute vascular injury. We will modulate this signaling axis using AdvSca1-SM-specific Gli1 overexpression or Klf4 knockout; the effects of these modifications on AdvSca1-SM maintenance or differentiation will be assessed using immunofluorescent microscopy, flow cytometry, and single cell RNA-seq. Ultimately, the work proposed here represents a novel avenue of research that will help fill the current knowledge gap regarding the role AdvSca1-SM cells play in the atherosclerotic disease process.

项目摘要/摘要 动脉粥样硬化是一种复杂的炎症性疾病，是全球发病率和死亡率的主要原因，但 当前的疗法无法充分满足临床需求。直到最近，大多数动脉粥样硬化研究一直集中在 内皮激活，免疫细胞募集和脂质保留率，但有新兴的证据 植入动脉粥样硬化的发病机理中的血管外膜。具体来说，有人建议 冒险微血管的扩展，Vasa vasorum（VV），通过促进动脉粥样硬化进展 炎性细胞浸润到血管壁并发展动脉瘤。该项目着重于 平滑肌衍生的血管祖细胞，称为ADVSCA1-SM细胞，在VASA血管膨胀中发挥作用 和斑块新生血管形成。我们的实验室先前将Advsca1-SM细胞识别为居民血管的子集 祖细胞在KLF4依赖性过程中从成熟的平滑肌细胞进行重编程。这些细胞 在刺猬/Wnt/wnt/β-catenin途径（尤其是GLI1）中富含基因，并具有多曲目 分化潜力。在急性血管损伤的情况下，Advsca1-SM细胞稳健地扩展以贡献 要进行纤维化重塑，但目前尚不清楚它们在慢性损伤（例如动脉粥样硬化）中的作用。工作 几个小组表明，冒险祖细胞可以有助于VV扩展，我们的小组 以前已经证明ADVSCA1-SM细胞能够通过内皮或 Matrigel插头测定中的平滑肌细胞分化。此外，我们已经确定了Advsca1-SM样细胞 在人类冠状动脉周围的微丝中，表明在VV中具有潜在的作用。这个目的 项目旨在确定Advsca1-SM细胞对VV扩展，牙菌斑新生血管形成和 动脉粥样硬化进展。我们假设ADVSCA1-SM细胞是动脉粥样硬化的主要因素， 这些细胞的调节可以改变疾病进展。特定目标1将定义功能 Advsca1-SM细胞使用高度特定的advsca1-贡献对VV扩展和/或斑块的进展 SM遗传谱系追踪小鼠模型； ADVSCA1-SM贡献将使用免疫荧光评估 显微镜和流式细胞术。特定的目标2将定义控制ADVSCA1-SM细胞的机制 动脉粥样硬化的设置，重点是先前被识别为 在急性血管损伤的情况下，ADVSCA1-SM细胞的调节剂。我们将使用 Advsca1-SM特异性GLI1过表达或KLF4敲除；这些修改对ADVSCA1-SM的影响 将使用免疫荧光显微镜，流式细胞仪和单一评估维护或分化 细胞RNA-seq。最终，此处提出的工作代表了一种新颖的研究途径，这将有助于填补 当前有关ADVSCA1-SM细胞在动脉粥样硬化疾病过程中起作用的知识差距。