Regulation of Vascular Calcification by Adventitial Endothelial Cells

外膜内皮细胞对血管钙化的调节

基本信息

批准号：
10642619
负责人：
Xinjiang Cai
金额：
$ 18.71万
依托单位：
UNIVERSITY OF CALIFORNIA LOS ANGELES
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-06-01 至 2028-05-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10642619
关键词：
Acceleration Adult Affect Age Age Years Animal Model Aorta Apolipoprotein E Arterial Fatty Streak Arteries Atherosclerosis Blood Vessels Bone Development Cardiovascular system Cartilage Cell Line Cellular biology Chondrocytes Data Deposition Development Development Plans Disease Endothelial Cells Endothelium Exhibits Fluorescence Microscopy Foundations Gene Expression Gene Expression Profile Gene Targeting Glycoproteins Human K-Series Research Career Programs Knock-out Knockout Mice Knowledge Light Medical Mentors Mesenchymal Minerals Modeling Molecular Molecular Profiling Morbidity - disease rate Mus NOTCH1 gene Osteogenesis PECAM1 gene Pathologic Pathologic Processes Pathology Patients Pattern Phenotype Physicians Population Regulation Reporting Research Scientist Severities Signal Transduction Supporting Cell Therapeutic Tunica Adventitia Vascular Diseases Vascular Endothelial Cell Vascular calcification angiogenesis arterial stiffness biomineralization bone calcification cardiovascular risk factor career career development endothelial-specific sialomucin insight matrix Gla protein microCT microscopic imaging mortality notch protein novel novel therapeutic intervention osteogenic osteoprogenitor cell prevent receptor single-cell RNA sequencing stem cells transcriptome sequencing vascular contributions

Acceleration Adult Affect Age Age Years Animal Model Aorta Apolipoprotein E Arterial Fatty Streak Arteries Atherosclerosis Blood Vessels Bone Development Cardiovascular system Cartilage Cell Line Cellular biology Chondrocytes Data Deposition Development Development Plans Disease Endothelial Cells Endothelium Exhibits Fluorescence Microscopy Foundations Gene Expression Gene Expression Profile Gene Targeting Glycoproteins Human K-Series Research Career Programs Knock-out Knockout Mice Knowledge Light Medical Mentors Mesenchymal Minerals Modeling Molecular Molecular Profiling Morbidity - disease rate Mus NOTCH1 gene Osteogenesis PECAM1 gene Pathologic Pathologic Processes Pathology Patients Pattern Phenotype Physicians Population Regulation Reporting Research Scientist Severities Signal Transduction Supporting Cell Therapeutic Tunica Adventitia Vascular Diseases Vascular Endothelial Cell Vascular calcification angiogenesis arterial stiffness biomineralization bone calcification cardiovascular risk factor career career development endothelial-specific sialomucin insight matrix Gla protein microCT microscopic imaging mortality notch protein novel novel therapeutic intervention osteogenic osteoprogenitor cell prevent receptor single-cell RNA sequencing stem cells transcriptome sequencing vascular contributions

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项目摘要

PROJECT SUMMARY/ABSTRACT Vascular calcification affects ~60% of adults over 60 years of age and is frequently seen in patients with atherosclerosis. Atherosclerotic calcification is an independent risk factor for cardiovascular morbidity and mortality. Despite decades of research, no medical therapy has been convincingly established to prevent or reverse vascular calcification. The intimal (luminal) endothelium forms the innermost layer of the vasculature. Under disease conditions, the intimal endothelial cells (ECs) are stimulated to transform into osteoprogenitor cells through endothelial-mesenchymal transitions and contribute to vascular calcification. ECs in the adventitia contribute to neoangiogenesis in vascular disease. However, little is known about whether adventitial ECs are associated with vascular calcification. During bone development, a subset of ECs defined by high expression of the EC marker CD31 and the glycoprotein Endomucin (Emcn) are critical for bone formation. Our preliminary study using Matrix Gla Protein (Mgp) knockout (Mgp-/-) mice as a model of vascular calcification revealed two distinct populations of ECs in the calcified aortas, the intimal ECs (i-ECs) defined by CD31+Emcn- and the adventitial ECs (a-ECs) characterized by CD31+Emcn+. These two EC subtypes with distinct Emcn expression levels were also detected in human calcified arteries. Bulk RNA sequencing studies showed i-ECs were enriched in stem cell and osteogenic markers and a-ECs exhibited upregulated Notch expression. Endothelial deletion of the Notch1 gene reduced vascular calcification and increased the survival of the Mgp-/- mice. In this proposal, we hypothesize that a-ECs support the biomineralization in the vascular calcification in animal models and human atherosclerotic lesions, and are closely regulated by Notch signaling. In Aim 1, we will define the molecular signature of the a-EC (CD31+Emcn+) population in vascular calcification using single cell RNA sequencing. We will delineate the developmental trajectories of a-ECs in vascular calcification with correlation to the extent and severity of calcification. In Aim 2, we will determine the contribution of endothelial subtypes (a- ECs and i-ECs) and Notch signaling in atherosclerotic calcification. We will investigate the effect of endothelial- specific deletion of the Notch1 receptor on EC subtypes, calcification and transcriptional profiles in atherosclerotic lesions. The proposed studies will provide novel insight into the fundamental mechanisms of endothelial cell biology in atherosclerotic calcification and may identify potential gene targets for selective therapeutic modulation. Together with the mentored career development plan, the completion of the projects will serve as a foundation to facilitate the candidate to transition into a successful and independent physician scientist in cardiovascular research.

项目摘要/摘要血管钙化影响60岁以上的成年人的约60％，经常在患有动脉粥样硬化。动脉粥样硬化钙化是心血管发病率和死亡。尽管进行了数十年的研究，但仍未建立任何医疗疗法来预防或反向血管钙化。内皮（腔）内皮形成脉管系统的最内向层。在疾病条件下，刺激内皮内皮细胞（EC）以转化为骨基因因细胞通过内皮间充质转变，导致血管钙化。 ECS中的ECS 在血管疾病中有助于新血管生成。但是，关于外在EC是否是与血管钙化相关。在骨发育过程中，由高表达的EC的子集 EC标记CD31和糖蛋白内核素（EMCN）对于骨形成至关重要。我们的初步使用基质GLA蛋白（MGP）基因敲除（MGP - / - ）小鼠作为血管钙化模型的研究发现了两种钙化的主动脉，CD31+EMCN-定义的内膜EC（I-EC）和以CD31+EMCN+为特征的Adventitial ECS（A-ECS）。这两个具有不同EMCN表达的EC亚型在人钙化动脉中也检测到水平。散装RNA测序研究表明I-EC富含在干细胞和成骨标记和A-EC中表现出上调的凹口表达。内皮删除 Notch1基因降低了血管钙化并增加了MGP - / - 小鼠的存活率。在此提案中，我们假设A-ECS支持动物模型中血管钙化中的生物矿化，并支持人动脉粥样硬化病变，并受到Notch信号的密切调节。在AIM 1中，我们将定义使用单细胞RNA，血管钙化中A-EC（CD31+EMCN+）种群的分子特征测序。我们将描述与相关性血管钙化中A-EC的发育轨迹在钙化的范围和严重程度上。在AIM 2中，我们将确定内皮亚型的贡献（A- 动脉粥样硬化钙化中的ECS和I-ECS和Notch信号传导。我们将研究内皮的影响在EC亚型，钙化和转录曲线上的Notch1受体的特定缺失在动脉粥样硬化病变。拟议的研究将提供对有关的新见解动脉粥样硬化钙化中的内皮细胞生物学治疗调节。与指导的职业发展计划一起，项目的完成将作为促进候选人过渡为成功和独立医师科学家的基础在心血管研究中。