Modulation of Endothelial Cell Function by the Shear Stress-Responsive miR-155

剪切应力响应性 miR-155 对内皮细胞功能的调节

• 批准号: 8162633
• 负责人: CHARLES D SEARLES
• 金额: $ 32.28万
• 依托单位: EMORY UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-08-01 至 2015-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8162633
• 关键词: Actins; Address; Adhesions; Angiotensin II Receptor; Anti-Inflammatory Agents; Anti-inflammatory; Antiatherogenic; Aorta; Apoptosis; Arterial Fatty Streak; Atherosclerosis; Biological Process; Blood Platelets; Blood Vessels; Blood flow; Brain; Cell Culture Techniques; Cell physiology; Cells; Cellular Morphology; Cellular biology; Coronary; Critical Pathways; Cytoskeleton; Data; Development; Diabetes Mellitus; Differentiation and Growth; Disease; Endothelial Cells; Etiology; Functional disorder; Gene Expression; Gene Expression Regulation; Gene Targeting; Genes; Heart Diseases; Hematopoiesis; Hematopoietic; Homeostasis; Hour; Human; Hypertension; Inflammatory; Knowledge; Leukocytes; Lymphocyte; Mediating; MicroRNAs; Modeling; Molecular; Molecular Biology; Morphogenesis; Mus; Pathway interactions; Pattern; Permeability; Phenotype; Play; Process; Production; Publishing; Regulation; Regulator Genes; Regulatory Pathway; Relative (related person); Research; Rho-associated kinase; Risk Factors; Role; Signal Pathway; Signal Transduction; Skeletal Muscle; Smoking; Smooth Muscle Myocytes; Stimulus; Testing; Trees; Umbilical vein; Untranslated RNA; Vascular Diseases; Vascular remodeling; Work; angiogenesis; atheroprotective; blood pressure regulation; cardiogenesis; cell motility; cell type; gene function; gene interaction; hypercholesterolemia; in vivo; insight; member; migration; monolayer; mortality; mouse model; myo-inositol-1 (or 4)-monophosphatase; overexpression; receptor expression; response; shear stress; tumorigenesis

DESCRIPTION (provided by applicant): 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 posttranscriptional 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 one highly shear-responsive miRNA, miR-155, will be studied. Specifically, the proposed research will define the impact of miR-155-target gene interaction on endothelial cell apoptosis, barrier function and migration. To study the mechanism by which miR-155 modulates apoptosis, we will focus on the SHIP1/PI3K/Akt pathway. To study the mechanism by which miR-155 modulates endothelial monolayer permeability and migration, we will focus on the RhoA/Rho kinase pathway. We will test the influence of miR- 155 on these critical pathways by experimentally manipulating expression of miR-155, 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, monolayer permeability, and migration will be quantified. Finally, the association between shear-induced miR-155 expression and activity of endothelial cell regulatory pathways 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. PUBLIC HEALTH RELEVANCE: Coronary atherosclerotic heart disease is an inflammatory disease that is the greatest cause of mortality in the U.S. Although the etiology of atherosclerosis is related to risk factors, such as diabetes, hypertension, hypercholesterolemia, and smoking, the inflammatory process occurs preferentially in arterial regions associated with low and disturbed blood flow while sparing the undisturbed flow regions, indicating that blood flow patterns are essential to the disease process. The purpose of this project is to examine the mechanisms by which an important new class of regulatory molecules, known as microRNAs, modulate vascular gene expression and function in response to flow.

描述（由申请人提供）：该提案的目的是定义microRNA表达对响应剪切应力力发生的内皮细胞功能变化的影响。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），单链，非编码的RNA，它们已成为分子生物学研究的主要重点，因为它们的转录后调节了涉及一系列细胞功能的基因的表达，包括分化，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，将研究一种高度剪切反应性miRNA，miRNA，miRNA，miR-155。具体而言，拟议的研究将定义miR-155靶基因相互作用对内皮细胞凋亡，屏障功能和迁移的影响。为了研究miR-155调节细胞凋亡的机制，我们将专注于Ship1/pi3k/akt途径。为了研究miR-155调节内皮单层渗透性和迁移的机制，我们将重点关注RhoA/Rho激酶途径。我们将通过通过实验操纵miR-155的表达，其靶基因或途径的成员在MiRNA-Target基因相互作用的下游来测试miR-155对这些关键途径的影响。随后，将量化这些操纵对内皮细胞凋亡，单层渗透性和迁移的影响。最后，将在主动脉流动的小鼠模型中研究剪切诱导的miR-155表达与内皮细胞调节途径的活性之间的关联。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。 PUBLIC HEALTH RELEVANCE: Coronary atherosclerotic heart disease is an inflammatory disease that is the greatest cause of mortality in the U.S. Although the etiology of atherosclerosis is related to risk factors, such as diabetes, hypertension, hypercholesterolemia, and smoking, the inflammatory process occurs preferentially in arterial regions associated with low and disturbed blood flow while sparing the undisturbed flow regions, indicating that blood flow patterns are对疾病过程至关重要。该项目的目的是检查一种重要的新型调节分子（称为microRNAS）调节血管基因表达和功能对流动的功能的机制。