PARs and S1P receptors in endothelial biology

内皮生物学中的 PAR 和 S1P 受体

• 批准号: 8279302
• 负责人: SHAUN R. COUGHLIN
• 金额: $ 55.81万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-09-30 至 2014-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8279302
• 关键词: Adult; Affect; Allergic Reaction; Anaphylaxis; Anatomy; Apical; Atherosclerosis; Autocrine Communication; Basement membrane; Biology; Blood Platelets; Blood Vessels; Cardiovascular system; Cell Culture Techniques; Cell Line; Cell membrane; Cell physiology; Cells; Coagulation Process; Complex; Development; Disease; Embryo; Embryonic Development; Employee Strikes; Endothelial Cells; Exhibits; Extravasation; Failure; Functional disorder; G Protein-Coupled Receptor Genes; G-Protein-Coupled Receptors; Genes; Genetic; Goals; Growth; Health; In Vitro; Inflammatory; Injury; Knockout Mice; Laboratories; Leukocytes; Ligation; Long-Term Effects; Maintenance; Malignant Neoplasms; Metabolic; Metabolism; Modeling; Molecular; Mus; Myocardial Infarction; Patients; Peptide Hydrolases; Permeability; Phosphotransferases; Physiology; Plasma; Play; Property; Proteinase-Activated Receptors; Receptor Activation; Receptor Signaling; Regulation; Role; Sepsis; Signal Pathway; Signal Transduction; Source; Specificity; Sphingosine; Sphingosine-1-Phosphate Receptor; Stroke; Surface; System; Testing; Thrombus; Tissues; Toxic effect; Transgenic Organisms; Vascular Permeabilities; base; cell type; detector; edg-1 Protein; in vivo; monolayer; mouse model; receptor; receptor function; response; sphingosine 1-phosphate; sphingosine kinase

DESCRIPTION (provided by applicant): We will characterize two signaling systems that regulate the permeability and integrity of blood vessels: coagulation proteases and protease-activated receptors, and sphingosine-1-phosphate and S1P receptors. We will test the hypothesis that both systems sense extravasation of plasma and trigger appropriate endothelial cell responses, and we will explore parallels and possible connections between these systems. We shall ask: 1) How is S1P important for regulation of vascular permeability and integrity? We generated adult mice that fail to supply S1P to plasma and found striking abnormalities in vascular permeability and integrity. We shall determine a) whether altered barrier function in these "pS1Pless" mice is due to failure to metabolize sphingosine with consequent endothelial cell-autonomous metabolic/toxic effects or to failure to supply S1P to plasma and S1P receptor activation on endothelial and other cells, b) the anatomic basis for their increased vascular leak and whether it shows tissue or vessel-type specificity, c) whether endothelial cells are the main target of plasma S1P signaling in this context, and if so, whether such signaling is continuous or whether plasma S1P provides a dynamic signal that enables endothelial cells to sense and help terminate leaks, and d) the long-term effects of lack of plasma S1P and whether they are due to dysregulated barrier function. 2) How are PARs important for regulation of vascular permeability and integrity? We generated knockout mice for all the PARs and relevant transgenics and will use these to determine a) the effects of activation of different endothelial PARs on vascular permeability and integrity in vivo and whether pS1Pless mice provide a sensitized system for uncovering such roles for PARs, b) whether PAR signaling is parallel to, partially redundant with, or dependent upon S1P signaling. 3) Do differences in apical and basal S1P and PAR function contribute to their roles in barrier regulation? Our preliminary studies suggest a model that would permit S1P signaling to serve a dynamic leak detector function and raise new questions regarding PARs by analogy. We will determine a) whether endothelial cell S1P1 receptors display apical-basal polarity in vitro and in vivo to enable the dynamic leak detecting function we posit, and b) whether apical and basal differences, such as EPCR ligation, modulate the effects of PAR activation on either surface. Complementary genetic and pharmacological approaches will be used in mouse models and in cell culture. Preliminary studies reveal an important role for plasma S1P in regulating endothelial barrier function in vivo, distinct barrier responses to activation of different PARs, dramatic effects of manipulation of S1P and PAR signaling on survival in models of anaphylaxis, and long-term effects of altered barrier function in vivo. The proposed studies will provide new information regarding vascular physiology and pathophysiology. PUBLIC HEALTH RELEVANCE: Our studies will reveal new molecular and cellular mechanisms by which the endothelial cells that line blood vessels sense and regulate leakiness. Alterations in endothelial permeability play an important role in allergic reactions, new blood vessel growth in cancer, heart attacks and strokes, atherosclerosis, and inflammatory states including sepsis. Thus, understanding of how to manipulate the endothelial leakiness might benefit patients with a range of disorders.

描述（由申请人提供）：我们将表征两个调节血管渗透性和完整性的信号系统：凝血蛋白酶和蛋白酶激活的受体，以及鞘氨醇-1-磷酸盐和S1P受体。我们将检验以下假设：两个系统都感觉到血浆的渗出和触发适当的内皮细胞反应，我们将探索这些系统之间的相似之处和可能的连接。我们将问：1）S1P对于调节血管渗透性和完整性的重要性至关重要吗？我们产生了未能向等离子体提供S1P的成年小鼠，并发现血管渗透性和完整性异常。我们应确定a）这些“ PS1PLESS”小鼠中的屏障功能的改变是由于未能代谢鞘氨酸而导致的，因此内皮细胞自治的代谢/有毒作用或未能向血浆和S1P受体激活提供S1P，以及其他细胞上的细胞和其他细胞的基础，它是否显示出渗透性，b）它是否显示出渗透性，b）b）b）b）b）b）b）b）b）b）b）b）在这种情况下，内皮细胞是血浆S1P信号传导的主要目标，如果是的，无论是连续的信号传导还是等离子体S1P是否提供了动态信号，使内皮细胞能够感知并有助于终止泄漏，d）d）缺乏血浆S1P的长期影响以及它们是否归因于障碍障碍。 2）PARS对于调节血管渗透性和完整性的重要性很重要吗？我们为所有分型和相关的转基因生成了敲除小鼠，并将使用这些小鼠来确定不同内皮par激活对体内血管渗透性和完整性的影响，以及PS1Pless小鼠是否提供了一个敏感的系统，用于发现此类pars的作用； 3）顶端和基础S1P和PAR功能的差异是否有助于它们在屏障调节中的作用？我们的初步研究提出了一个模型，该模型将允许S1P信号传导发挥动态泄漏检测器功能，并通过类比提出有关PARS的新问题。我们将确定a）内皮细胞S1P1受体在体外和体内表现出根尖的基础极性，以使动态泄漏检测功能我们提出，b）b）诸如EPCR连接之类的根尖和基础差异是否可以调节PAR激活对任一表面的影响。互补的遗传和药理方法将用于小鼠模型和细胞培养。初步研究揭示了血浆S1P在体内调节内皮屏障功能中的重要作用，对不同PAR的激活的明显屏障响应，S1P操纵的急剧影响和PAR信号对过敏反应模型的存活以及长期障碍物在体内的长期影响。拟议的研究将提供有关血管生理和病理生理学的新信息。公共卫生相关性：我们的研究将揭示新的分子和细胞机制，通过这些机制将血管缠绕并调节泄漏感的内皮细胞。内皮通透性的改变在过敏反应，癌症的新血管生长，心脏病发作和中风，动脉粥样硬化和包括败血症在内的炎症状态中起着重要作用。因此，了解如何操纵内皮泄漏可能会使患有多种疾病的患者受益。