The effect of laminar and disturbed flow on endothelial glucose metabolism

层流和扰动流对内皮葡萄糖代谢的影响

基本信息

批准号：
10335226
负责人：
Alisa S Morss Clyne
金额：
$ 38.29万
依托单位：
UNIV OF MARYLAND, COLLEGE PARK
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-01-01 至 2023-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10335226
关键词：
Acetyl Coenzyme A Actins Affect Animal Model Area Arterial Fatty Streak Atherosclerosis Blood Vessels Blood flow Caveolae Cell Adhesion Molecules Cell Proliferation Cells Cellular Metabolic Process Collaborations Data Development Disease Endothelial Cells Endothelium Enzymes Epigenetic Process Exposure to Functional disorder Gene Expression Glucose Glycolysis Goals Golgi Apparatus Health Hexosamines Histone Acetylation Homeostasis Hour Impairment In Vitro Inflammation Inflammatory Learning Link Lipids Malignant Neoplasms Mass Spectrum Analysis Mediating Mediator of activation protein Metabolic Metabolic Pathway Metabolism Modeling Morphology Myocardial Infarction NOS3 gene Neoplasm Metastasis Nitric Oxide Oxygen Pathologic Pathway interactions Permeability Phenotype Phosphorylation Post-Translational Protein Processing Production Proteins Regulation Repression Research Research Personnel Side Stress Fibers Stroke Testing Time Vascular remodeling Vasodilation Warburg Effect angiogenesis atheroprotective cancer cell career endothelial dysfunction experience experimental study glucose metabolism glucose uptake hemodynamics in vivo metabolomics novel therapeutics overexpression prevent shear stress therapy development tool

项目摘要

Endothelial metabolism has recently re-emerged as a powerful tool to regulate vascular function. However, studies have focused entirely on glycolytic flux regulation via PFKFB3 and its effects in angiogenesis. Little is known about how endothelial cell metabolism impacts macrovascular endothelial function in health and disease. Endothelial cells are constantly exposed to shear stress from the flowing blood. Endothelial cells in steady laminar flow express a quiescent phenotype, maintaining vascular homeostasis through control of proliferation, permeability, inflammation, and vascular tone. Endothelial cells in oscillating disturbed flow express an athero- prone phenotype with elevated proliferation, permeability, and inflammatory adhesion molecule expression as well as impaired NO production (defined as endothelial dysfunction). Disturbed flow regions are linked to subsequent pathological vascular remodeling including atherosclerotic plaque development. Recently, endothelial cells in steady laminar reduced glycolysis partially via KLF2-mediated repression of PFKFB3. However, concurrent KLF2 and PFKFB3 overexpression did not fully restore glycolytic rate, suggesting that other metabolic mediators are involved. Our data show that endothelial cells in steady laminar flow reduce glycolytic flux at shorter times with no change in PFKFB3 expression, and that endothelial cells in oscillating disturbed flow do not decrease glycolytic flux. Our data also show that flow regulates the hexosamine biosynthetic pathway, a side branch of glycolysis which controls protein O-GlcNAcylation, and acetyl CoA, which is critical to lipid synthesis and histone acetylation. We are only beginning to discover mechanisms by which shear stress affects endothelial glucose metabolism and downstream pathways. Our long term goal is to modulate glucose metabolism to reduce endothelial dysfunction in disturbed flow. The goal of this project is to understand how steady laminar and oscillating disturbed flow differentially affect macrovascular endothelial glycolytic flux, the HBP, and acetyl CoA metabolism. We hypothesize that mean shear stress greater than 12 dynes/cm2 reduces glycolytic flux, eNOS O-GlcNAcylation, and acetyl CoA to promote an athero-protective endothelial phenotype. To test this hypothesis, we will (1) determine how steady laminar and oscillating disturbed flow regulate endothelial glycolytic flux; (2) determine how steady laminar and oscillating disturbed flow affect eNOS O-GlcNAcylation; and (3) determine how altered acetyl CoA in flow impacts lipid synthesis and histone acetylation Since atherosclerosis is a disease of altered metabolism, we will use a combination of in vitro and ex vivo experiments to discover mechanisms underlying changes in glucose metabolism with flow. Our team is uniquely prepared to pursue this research, with expertise in endothelial hemodynamics, metabolic mass spectrometry, O- GlcNAcylation, and ex vivo vessel analysis. These data will transform the field by creating a new research area at the intersection of hemodynamics and metabolomics.

内皮代谢最近重新出现是调节血管功能的强大工具。然而，研究完全集中于通过PFKFB3及其在血管生成中的影响的糖酵解通量调节。几乎没有知道内皮细胞代谢如何影响健康和疾病中的大血管内皮功能。内皮细胞不断暴露于流动血液中的剪切应力。稳定的内皮细胞层流表达静止的表型，通过控制增殖，维持血管稳态渗透性，炎症和血管张力。振荡流动流动的内皮细胞表达动脉粥样硬化俯卧的表型，具有升高，渗透性和炎症粘附分子表达升高为以及没有生产受损（定义为内皮功能障碍）。流动区域被干扰与随后的病理血管重塑，包括动脉粥样硬化斑块发育。最近，稳定层流中的内皮细胞通过KLF2介导的抑制部分降低了糖酵解 PFKFB3。但是，并发KLF2和PFKFB3过表达并未完全恢复糖酵解速率，表明涉及其他代谢介质。我们的数据表明，内皮细胞稳定层流在较短的时间减少糖酵解通量，而PFKFB3表达没有变化，而内皮振荡流动流动的细胞不会降低糖酵解通量。我们的数据还表明，流量调节己胺生物合成途径，糖酵解的侧支，控制蛋白O-glcnacylation和乙酰基 COA，这对于脂质合成和组蛋白乙酰化至关重要。我们才开始发现剪切应力影响内皮葡萄糖代谢和下游途径的机制。我们的长期目标是调节葡萄糖代谢，以减少流动干扰的内皮功能障碍。该项目的目的是了解稳定的层流和振荡流动的差异影响大血管内皮糖酵解通量，HBP和乙酰COA代谢。我们假设这意味着剪切大于12 dynes/cm2的应力可减少糖酵解通量，eNOS O-glcnacylation和乙酰基COA，以促进动脉保护性内皮表型。为了检验这一假设，我们将（1）确定层流和振荡流动的流动调节内皮糖酵解通量；（2）确定层流的稳定性和振荡流动的流动会影响eNOS O-Glcnacylation；（3）确定乙酰基COA如何改变流量会影响脂质合成和组蛋白乙酰化由于动脉粥样硬化是一种新陈代谢的疾病，我们将使用体外和离体的组合实验以发现随着流动的葡萄糖代谢发生变化的机制。我们的团队是独特的准备从事这项研究，并具有内皮血液动力学，代谢质谱法，O-的专业知识 Glcnacylation和离体血管分析。这些数据将通过创建一个新的研究领域来改变现场在血液动力学和代谢组学的交集中。