PARs and S1P receptors in endothelial biology

内皮生物学中的 PAR 和 S1P 受体

• 批准号: 7905814
• 负责人: SHAUN R. COUGHLIN
• 金额: $ 56.37万
• 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 2014-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7905814
• 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; In Vitro; Inflammatory; Injury; Knockout Mice; Laboratories; Leukocytes; Ligation; Long-Term Effects; Maintenance; Malignant Neoplasms; Metabolic; Metabolism; Modeling; Molecular; Mus; Myocardial Infarction; PAR-2 Receptor; 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; public health relevance; 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）：我们将表征调节血管通透性和完整性的两种信号系统：凝血蛋白酶和蛋白酶激活受体，以及 1-磷酸鞘氨醇和 S1P 受体。我们将测试两个系统都能感知血浆外渗并触发适当的内皮细胞反应的假设，并且我们将探索这些系统之间的相似之处和可能的联系。我们会问：1）S1P 对于血管通透性和完整性的调节有何重要意义？我们培育了无法向血浆供应 S1P 的成年小鼠，并发现血管通透性和完整性存在显着异常。我们将确定 a) 这些“pS1Pless”小鼠中屏障功能的改变是否是由于未能代谢鞘氨醇而导致内皮细胞自主代谢/毒性作用，或者是由于未能向血浆供应 S1P 以及内皮细胞和其他细胞上的 S1P 受体激活， b) 血管渗漏增加的解剖学基础以及是否显示组织或血管类型特异性，c) 内皮细胞是否是血浆的主要目标在这种情况下，S1P 信号传导，如果是，这种信号传导是否是连续的，或者血浆 S1P 是否提供动态信号，使内皮细胞能够感知并帮助终止渗漏，以及 d) 缺乏血浆 S1P 的长期影响以及它们是否是由于屏障功能失调所致。 2) PAR 对于调节血管通透性和完整性有何重要意义？我们生成了所有 PAR 和相关转基因的敲除小鼠，并将使用它们来确定 a) 不同内皮 PAR 的激活对体内血管通透性和完整性的影响，以及 pS1Pless 小鼠是否提供了一个敏化系统来揭示 PAR 的此类作用，b ) PAR 信令是否与 S1P 信令并行、部分冗余或依赖于 S1P 信令。 3）顶端和基底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种模型，该模型允许 S1P 信号传递动态检漏仪功能，并通过类比提出有关 PAR 的新问题。我们将确定 a) 内皮细胞 S1P1 受体是否在体外和体内表现出顶端-基底极性，以实现我们假设的动态泄漏检测功能，以及 b) 顶端和基底差异（例如 EPCR 连接）是否调节 PAR 激活的影响在任一表面上。互补的遗传和药理学方法将用于小鼠模型和细胞培养。初步研究揭示了血浆 S1P 在调节体内内皮屏障功能中的重要作用、对不同 PAR 激活的不同屏障反应、操纵 S1P 和 PAR 信号传导对过敏反应模型生存的显着影响，以及屏障改变的长期影响在体内发挥作用。拟议的研究将提供有关血管生理学和病理生理学的新信息。公共健康相关性：我们的研究将揭示血管内皮细胞感知和调节渗漏的新分子和细胞机制。内皮通透性的改变在过敏反应、癌症中的新血管生长、心脏病和中风、动脉粥样硬化以及包括脓毒症在内的炎症状态中发挥着重要作用。因此，了解如何控制内皮渗漏可能有益于患有一系列疾病的患者。