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.

项目概要/摘要慢性炎症和循环胆固醇升高引起的内皮功能障碍会促进导致冠心病的主要动脉内皮下斑块的形成修复受损的内皮细胞具有巨大的治疗前景。心脏病；然而，内源性内皮细胞（EC）再生是一个低效的过程。恢复动脉内皮的通畅将提供显着的治疗进展。血管平滑肌细胞 (VSMC) 构成动脉壁中的大部分细胞，能够响应病理生理刺激的表型可塑性，这些细胞代表了一个有吸引力的来源揭示功能性内皮细胞的分子机制和信号通路。将 VSMC 分化为 EC 以修复受损的内皮细胞将建立一种新的治疗模式我们的长期目标是发现新的分子和信号通路促进 VSMC 向内皮细胞的转变 (MEndoT) 我们的实验室已鉴定并表征了一系列的特征。进化上保守的内吞接头蛋白，称为 epsins，在协调我们的研究表明 EC 和骨髓细胞中 Epsins 1 和 2 的丢失。减少血管炎症并防止斑块的发生和进展。靶向驱动病变进展以及斑块组成的细胞中的epsins的治疗效果稳定性，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我们建议探究 VSMC epsin 蛋白在这些过程中的功能，并确定这一点。靶向这些蛋白质将促进有益的治疗性 VSMC 表型转换。初步研究表明，VSMC epsins 缺陷的 ApoE-/- 小鼠的斑块大小，增强斑块稳定性（包括纤维帽面积和斑块内 ACTA2+ 细胞的增加） cap），浸润细胞（CD45+免疫和炎症细胞以及CD68+泡沫）数量减少此外，RNA-seq 分析显示血管硬度和钙化显着减少。 Klf4，控制 VSMC 表型转换的多能转录因子，被下调 epsin 损失，以及 oxLDL 触发的 Runx2 泛素化和降解。根据这些发现，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具体目标：1) 确定epsins调节表型转换和间质转化的分子机制内皮分化，2) 确定epsins调节VSMC的分子机制成骨并促进动脉僵硬度，3) 确定靶向 epsins 的治疗潜力如果取得成果，拟议的研究将补充我们之前的工作和解决方案。强化了epsin蛋白可能作为冠心病有效治疗靶点的概念。