The Impact of miR-21 Expression on Endothelial Cell Apoptosis and Inflammation

miR-21表达对内皮细胞凋亡和炎症的影响

• 批准号: 8391582
• 负责人: CHARLES D SEARLES
• 金额: --
• 依托单位: VETERANS HEALTH ADMINISTRATION
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-10-01 至 2014-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8391582
• 关键词: Address; Adhesions; Anti-Inflammatory Agents; Anti-inflammatory; Antiatherogenic; Aorta; Apoptosis; Apoptotic; Arterial Fatty Streak; Atherosclerosis; Biological Process; Blood Vessels; Blood flow; Brain; Cell Adhesion Molecules; Cell physiology; Cells; Cellular Morphology; Cellular biology; Coronary; Data; Development; Diabetes Mellitus; Differentiation and Growth; Disease; Endothelial Cells; Etiology; Gene Expression; Gene Targeting; Genes; Heart Diseases; Hematopoietic; Hour; Human; Hypertension; Inflammation; Inflammatory; Inhibition of Apoptosis; Intercellular adhesion molecule 1; Knowledge; Mediating; MicroRNAs; Molecular; Molecular Biology; Morbidity - disease rate; Morphogenesis; PTEN gene; Pathway interactions; Pattern; Phenotype; Play; Population; Process; Regulation; Relative (related person); Research; Risk Factors; Role; Signal Pathway; Signal Transduction; Skeletal Muscle; Smoking; Stimulus; Testing; Trees; Untranslated RNA; Vascular Diseases; Vascular remodeling; Veterans; angiogenesis; atheroprotective; cardiogenesis; gene function; gene interaction; hypercholesterolemia; in vivo; insight; member; migration; monocyte; mortality; mouse model; public health relevance; research study; response; shear stress; therapeutic target; tumorigenesis

DESCRIPTION (provided by applicant): Project Summary The purpose of this proposal is to define the influence of microRNA expression on specific changes in endothelial cell function that occur in response to shear stress forces. Shear stress forces, generated by blood flow, play an important role in the regulation of vascular tone, vascular remodeling, and the focal development of atherosclerotic lesions. In the arterial tree, endothelial cells are exposed to different shear stress forces that induce distinct effects on gene expression and function. Unidirectional shear stress, which occurs in the straight part of the tree, elicits a change in endothelial gene expression that is generally anti-inflammatory and atheroprotective. In contrast, oscillatory shear stress, which occurs at branch points in the arterial tree, induces an overall pro-inflammatory and proatherosclerotic response. MicroRNAs (miRNAs) are a recently recognized class of short (19-25 nt), single stranded, noncoding RNAs that have become a major focus in molecular biology research because they posttranscriptionally regulate the expression of genes involved in an array of cell functions, including differentiation, growth, proliferation, and apoptosis. Although an important role for miRNA expression has been demonstrated for various biological processes, including cardiogenesis and angiogenesis, data on the role of specific miRNAs in endothelial cell biology is currently limited. In preliminary studies of human endothelial cells subjected to prolonged unidirectional shear stress (24 hrs, 15 dynes/cm2), a group of miRNAs was identified whose expression was significantly upregulated in response to this stimulus, suggesting that these miRNAs are important in regulating gene expression and function in endothelial cells. To further define the role of miRNA expression in modulating shear stress-induced changes in endothelial cell biology, the function of the most highly regulated miRNA, miR-21, will be studied. Specifically, the proposed experiments will define the impact of miR-21-target gene interaction on shear stress-induced changes in apoptosis and inflammatory molecule expression. Studies will be performed on cultured human endothelial cells subjected to unidirectional and oscillatory shear stress. The activity of apoptotic or inflammation signaling pathways containing miR-21 target genes, such as PI3K/Akt and MAP2K3, will be assessed. Pathway involvement will be tested by experimentally manipulating expression of miR-21, its target gene, or members of the pathway that are downstream of the miRNA-target gene interaction. Subsequently, the effect of these manipulations on endothelial cell apoptosis and adhesion molecule expression will be quantified. Finally, the interaction between miR-21 expression and apoptotic or inflammatory pathway activity will be studied in vivo, in a mouse model of altered aortic flow. We anticipate that these studies will help address a deficit in our knowledge about the function of miRNAs in endothelial cells and will enhance our understanding of the mechanisms by which shear stress forces modulate vascular disease.

描述（由申请人提供）： 项目摘要本提案的目的是定义microRNA表达对响应剪切应力力而发生的内皮细胞功能特定变化的影响。由血流产生的剪切应力力在调节血管张力，血管重塑和动脉粥样硬化病变的局灶性发展中起重要作用。在动脉树中，内皮细胞暴露于不同的剪切应力力，对基因表达和功能产生明显影响。发生在树的直部分中的单向剪切应力会引起通常具有抗炎和动脉保护性的内皮基因表达的变化。相比之下，发生在动脉树的分支点处的振荡剪应力会诱导整体促炎和促进性肌动膜反应。 microRNA（miRNA）是最近公认的短（19-25 nt）类，单链，非编码的RNA，它们已成为分子生物学研究的主要重点，因为它们在转录后调节了涉及一系列细胞功能的基因的表达，包括差异，生长，增殖，增殖和凋亡。尽管已经在包括心脏生成和血管生成在内的各种生物学过程中证明了miRNA表达的重要作用，但目前限制了有关特定miRNA在内皮细胞生物学中作用的数据。在对长期单向剪切应力（24小时，15 Dynes/cm2）的人内皮细胞的初步研究中，鉴定出一组miRNA，其对这种刺激的表达显着上调，这表明这些miRNA在调节基因表达和在内皮细胞中的作用很重要。为了进一步定义miRNA表达在调节剪切应力诱导的内皮细胞生物学变化中的作用，将研究最高度调节的miRNA，miRNA，miRNA的功能。具体而言，提出的实验将定义miR-21靶基因相互作用对剪切应力诱导的凋亡和炎症分子表达变化的影响。研究将对受到单向和振荡剪切应力的培养的人内皮细胞进行。将评估包含miR-21靶基因（例如PI3K/AKT和MAP2K3）的凋亡或炎症信号通路的活性。途径参与将通过实验操纵miR-21的表达，其靶基因的表达或途径的成员，这些途径是miRNA-target基因相互作用的下游的途径成员。随后，将量化这些操纵对内皮细胞凋亡和粘附分子表达的影响。最后，将在主动脉流动的小鼠模型中研究miR-21表达与凋亡或炎症途径活性之间的相互作用。我们预计这些研究将有助于解决我们对miRNA在内皮细胞中功能的知识的赤字，并将增强我们对剪切应力调节血管疾病的机制的理解。