Molecular Mechanisms Governing Vascular Cell Function and Phenotype in Health and Disease

健康和疾病中控制血管细胞功能和表型的分子机制

基本信息

批准号：
10380102
负责人：
Hong Chen
金额：
$ 74.34万
依托单位：
BOSTON CHILDREN'S HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-04-01 至 2025-03-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10380102
关键词：
Adaptor Signaling Protein Apolipoprotein E Area Arterial Fatty Streak Arteries Atherosclerosis Attenuated Biochemical Blood Vessels Cardiovascular Diseases Cause of Death Cell Culture Techniques Cell physiology Cells Cholesterol Chronic Complement Coronary heart disease Data Development Diet Disease Endocytosis Endothelial Cells Endothelium Family Foam Cells Foundations Gene Expression Genes Goals Health Heart Diseases Homeostasis Human Immune In Vitro Infiltration Inflammation Inflammatory Knockout Mice Laboratories Lesion Light Mediating Mesenchymal Mission Modeling Molecular Molecular Target Morbidity - disease rate Mouse Strains Mus Mutant Strains Mice Myeloid Cells Osteogenesis PTPRC gene Pathogenesis Pathologic Phenotype Play Process Proteins Reagent Resolution Role Signal Pathway Signal Transduction Small Interfering RNA Smooth Muscle Myocytes Source Stimulus Testing Therapeutic Therapeutic Effect Time Transgenic Organisms Ubiquitination United States National Institutes of Health Vascular Diseases Vascular Smooth Muscle Work arterial stiffness atherogenesis calcification cell regeneration endothelial dysfunction endothelial repair epsin epsin 1 experimental study in vitro Model injured injury and repair innovation insight mortality multidisciplinary nanoparticle nanoparticle delivery next generation novel novel therapeutics osteogenic overexpression oxidized low density lipoprotein pluripotency prevent response siRNA delivery targeted treatment therapeutic target tool transcription factor transcriptome sequencing transdifferentiation translational potential vascular inflammation

项目摘要

PROJECT SUMMARY/ABSTRACT Endothelial dysfunction resulting from chronic inflammation and elevated circulating cholesterol promotes the formation of plaques in the sub-endothelium of major arteries causing coronary heart disease—a leading cause of morbidity and mortality worldwide. Repair of the injured endothelium holds great promise to treat heart disease; however, endogenous endothelial cell (EC) regeneration is an inefficient process. The ability to restore patency of the arterial endothelium would provide a significant therapeutic advancement. Because vascular smooth muscle cells (VSMCs) constitute the majority of cells in the arterial wall and are capable of phenotypic plasticity in response to pathophysiological stimuli, these cells represent an appealing source of functional endothelial cells. Unraveling the molecular mechanisms and signaling pathways that govern trans- differentiation of VSMCs into ECs to mend the injured endothelium would establish a novel treatment paradigm for coronary heart disease. Our long-term goal is to discover new molecules and signaling pathways that facilitate VSMC-to-endothelial transition (MEndoT). Our laboratory has identified and characterized a family of evolutionarily-conserved endocytic adaptor proteins called epsins, which have crucial roles in coordinating endocytosis and signal transduction. Our studies show that loss of epsins 1 and 2 in ECs and myeloid cells reduces vascular inflammation and prevents plaque initiation and progression. To further assess the therapeutic effects of targeting epsins in cells that drive lesion progression as well as plaque composition and stability, we will use recently created disease-specific mice harboring VSMC-specific deficiency of these epsins. We propose to interrogate the function of VSMC epsin proteins in these processes and establish that therapeutic targeting of these proteins will promote beneficial VSMC phenotype switching. So far, our preliminary studies indicate that ApoE-/- mice with a deficiency in VSMC epsins have a significant reduction in plaque size, enhanced plaque stability (including an increase in fibrous cap area and ACTA2+ cells within the cap), a reduction in the number of infiltrating cells (CD45+ immune and inflammatory cells and CD68+ foam cells), and a prominent decrease in vascular stiffness and calcification. In addition, RNA-seq analyses show that Klf4, the pluripotent transcriptional factor controlling phenotypic switching of VSMCs, is downregulated by epsin loss, as is oxLDL-triggered Runx2 ubiquitination and degradation. In light of these findings, we will investigate the following Specific Aims using unique mutant mice, in vitro models, and novel reagents: 1) To determine the molecular mechanisms by which epsins regulate phenotype switching and mesenchymal-to- endothelial differentiation, 2) To determine the molecular mechanisms by which epsins regulate VSMC osteogenesis and promote arterial stiffness, and 3) To determine the therapeutic potential of targeting epsins for atheroma formation and resolution. If fruitful, the proposed study will complement our prior work and strengthen the concept that epsin proteins may serve as a potent therapeutic target for coronary heart disease.

项目摘要/摘要慢性炎症和循环胆固醇引起的内皮功能障碍可促进在主要动脉的亚内皮中形成斑块，引起冠心病，这是一个领先的全球发病和死亡率的原因。受伤的内皮的维修有很大的保证心脏病;但是，内源性内皮细胞（EC）再生是一个效率低下的过程。能力恢复动脉内皮的通畅性将提供显着的热进步。因为血管平滑肌细胞（VSMC）构成动脉壁中的大多数细胞，能够对病理生理刺激的响应表型可塑性，这些细胞代表了吸引人的来源功能性内皮细胞。阐明控制反式的分子机制和信号通路将VSMC分化为EC以修补受伤的内皮将建立一种新的治疗范式用于冠状动脉疾病。我们的长期目标是发现新分子和信号通路促进VSMC到内皮过渡（Mendot）。我们的实验室已经确定并描述了一个家庭称为EPSIN的进化保存的内吞衔接蛋白，在协调中具有至关重要的作用内吞作用和信号转导。我们的研究表明，ECS和髓样细胞中Epsins 1和2的损失减少血管感染并防止牙菌斑的启动和进展。进一步评估靶向EPSIN在驱动病变进展以及斑块组成和的细胞中的治疗作用稳定性，我们将使用最近创建的具有VSMC特异性缺陷的疾病特异性小鼠 Epsins。我们建议在这些过程中询问VSMC Epsin蛋白的功能，并确定这一点这些蛋白质的治疗靶向将促进有益的VSMC表型切换。到目前为止，我们的初步研究表明，VSMC Epsins缺乏症的APOE - / - 小鼠的apoE - / - 小鼠的幅度显着降低斑块尺寸，增强的斑块稳定性（包括纤维帽面积的增加和Acta2+细胞内 CAP），减少浸润细胞的数量（CD45+免疫和炎症细胞以及CD68+泡沫细胞），血管刚度和钙化显着降低。此外，RNA-seq分析显示 KLF4是控制VSMC的表型转换的多能转录因子，被下调 Epsin损失，oxLDL触发的Runx2泛素化和降解也是如此。鉴于这些发现，我们将使用独特的突变小鼠，体外模型和新型试剂研究以下特定目的：1）确定epsins调节表型切换和间充质至 - 向 - 到 - 的分子机制内皮分化，2）确定EPSIN调节VSMC的分子机制成骨和促进动脉刚度，以及3）确定靶向Epsins的治疗潜力用于动脉粥样硬化和分辨率。如果富有成果，拟议的研究将完成我们以前的工作，并且增强了艾培蛋白蛋白可以成为冠心病的潜在治疗靶点的概念。