Crosstalk between S1P Receptor 1/S1P1 and P-Selectin/Selp in Regulation of Inflammatory Vascular Permeability

S1P 受体 1/S1P1 和 P-选择素/Selp 之间的串扰调节炎症血管通透性

• 批准号: 10871784
• 负责人: STEVEN M DUDEK
• 金额: $ 31.31万
• 依托单位: UNIVERSITY OF FLORIDA
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-09-20 至 2027-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10871784
• 关键词: Actins; Acute Lung Injury; Acute Respiratory Distress Syndrome; Alveolar; Binding; Biological; Black Populations; Blood Vessels; CD44 gene; COVID-19 pandemic; Circulation; Code; Cytoskeleton; DNA Methylation; DOCK1 gene; Data; Deubiquitinating Enzyme; Deubiquitination; Endothelial Cells; Endothelium; Exhibits; Extravasation; FDA approved; Family suidae; Floods; Focal Adhesions; G-Protein-Coupled Receptors; GTP Phosphohydrolase Activators; Genes; Genetic; Glycoproteins; Guanosine Triphosphate Phosphohydrolases; Immunoglobulins; Inflammatory; Intervention; Ligands; Ligation; Link; Liposomes; Lung; Mechanical Stress; Mediating; Membrane; Modeling; Morbidity - disease rate; Multiple Organ Failure; Mus; P-Selectin; P-selectin ligand protein; Permeability; Pre-Clinical Model; Proteins; Proto-Oncogene Proteins c-akt; Pulmonary Circulation; Rattus; Regulation; Research Personnel; Risk; Role; Selectins; Signal Pathway; Signal Transduction; Site; Small Interfering RNA; Sphingosine-1-Phosphate Receptor; Susceptibility Gene; Therapeutic; Thrombosis; UCHL1 gene; Vascular Endothelial Cell; Vascular Permeabilities; Weibel-Palade Bodies; activated protein C receptor; analog; antagonist; demethylation; effective therapy; efficacy evaluation; epigenetic regulation; in vivo Model; inhibitor; insight; lipid mediator; lung injury; microvesicles; mortality; nitration; novel; pharmacologic; porcine model; pre-clinical; predictive marker; promoter; receptor; response; rho GTP-Binding Proteins; sphingosine 1-phosphate; survival prediction; targeted treatment; therapeutic effectiveness; trafficking; transcription factor; ubiquitin isopeptidase

PROJECT SUMMARY: The COVID-19 pandemic has dramatically highlighted a distinct vascular endotype of ARDS, characterized by profound endothelial cell (EC) permeability, thrombosis and microangiopathy (1; 2), and important unmet needs such as the critical absence of any effective FDA-approved therapies to halt or reverse lethal vascular leak in ARDS/VILI. Loss of lung EC barrier integrity results in vascular leakage, alveolar flooding, and is a critical feature of ARDS multi-organ failure and mortality. Project #4 investigators have demonstrated that sphingosine-1-phos- phate (S1P), a multifunctional lipid mediator, and its analog, Tysiponate (TySIP), effectively reduce vascular leakage and inflammatory lung injury via ligation of S1PR1, a G-protein-coupled receptor highly expressed in lung ECs. S1PR1 ligation rapidly initiates a signaling cascade that reorganizes the EC cytoskeleton via Rac GTPase signaling, enhances junctional integrity, and decreases vascular permeability. In contrast, ligation of the S1PR3 receptor induces Rho GTPase signaling to the cytoskeleton to increase lung permeability. Furthermore, S1PR3 is released into circulating microvesicles in VILI- or LPS-exposed mice serving as a novel ARDS bi- omarker that predicts survival. Both S1PR1 and S1PR3 promoter activity, expression and downstream signaling are influenced by pathophysiologic levels of mechanical stress with selective promoter demethylation, and by promoter SNPs associated with ARDS risk. We identified SELPLG as a novel ARDS susceptibility gene in Blacks, which encodes P-selectin glycoprotein ligand 1 (PSGL1) that via P-selectin (Selp) binding is critically involved in PMN trafficking. Our exciting data suggest interconnected signaling of S1PR/S1PR and PSGL1/SELPLG in regulation of EC vascular leak. Given the major role of ROS in pulmonary vascular leak and PMN trafficking, Specific Aim #1 (SA #1), with strong Core B assistance, will characterize the role of ROS- sensing transcription factors (HIF-1α/2α, NRF2), promoter SNPs (predominant in Blacks) and DNA methylation sites in genetic/epigenetic regulation of S1PR1/S1PR3 and SELPLG/SELP. Leveraging compelling preliminary data and critical Core D support, SA #2 will examine the novel barrier-regulatory roles for AKT nitration, protein deubiquitination (UCHL1), the Rac GTPase activator DOCK1, and the actin-binding FA protein, lamellipodin, in S1PR1 and S1PR3 signaling. SA #3 will characterize the dual influences of S1PR1/S1PR3 on PSGL1 secretion and P-selectin membrane mobilization from EC Weibel-Palade bodies, on signaling to the EC cytoskeleton, and altered EC barrier responses. Leveraging unparalleled Core C expertise in preclinical rat and porcine LPS/VILI models, SA #4 will evaluate the therapeutic effectiveness of the TySIPosome encargoed with either S1PR3 siRNAs or an UCHL1 activator, (CD19H28N205S), and directly assess the efficacy of a competitive inhibitor of P-selectin-PSGL-1 binding, the TSGL-Ig biologic (Tandem P-Selectin Glycoprotein Ligand-Immunoglobulin). Thus, Project #4 studies will address the unmet need for novel insights into ARDS-associated increases in EC permeability and for novel ARDS therapies that target S1P1/3/PSGL1 to restore integrity of the lung circulation.

项目概要： COVID-19大流行戏剧性地凸显了ARDS独特的血管内皮型，其特征为 严重的内皮细胞（EC）渗透性、血栓形成和微血管病（1; 2），以及重要的未满足需求 例如，严重缺乏任何有效的FDA批准的治疗方法来阻止或逆转致命的血管渗漏， ARDS/VILI。肺EC屏障完整性的丧失会导致血管渗漏、肺泡淹没，这是一个关键特征 多器官功能衰竭和死亡率的关系。项目#4研究人员已经证明，鞘氨醇-1-磷酸- 磷酸盐（S1 P），一种多功能脂质介质，及其类似物Tysiponate（TySIP），有效地减少血管 通过连接S1 PR 1（一种G蛋白偶联受体， 肺内皮细胞S1 PR 1连接快速启动信号级联，通过Rac重组EC细胞骨架 GT3信号传导，增强连接完整性，并降低血管通透性。相反， S1 PR 3受体诱导Rho GT3信号传导至细胞骨架以增加肺通透性。此外，委员会认为， S1 PR 3被释放到VILI或LPS暴露小鼠的循环微泡中，作为一种新的ARDS双 一种预测生存的标志物。S1 PR 1和S1 PR 3启动子活性、表达和下游信号传导 受机械应力的病理生理水平和选择性启动子去甲基化的影响， 与ARDS风险相关的启动子SNP。我们将SELPLG鉴定为一种新的ARDS易感基因， 布莱克，编码P-选择素糖蛋白配体1（PSGL 1），通过P-选择素（Selp）结合， 参与PMN贩卖我们令人兴奋的数据表明，S1 PR/S1 PR相互关联的信号传导， PSGL 1/SELPLG在EC血管渗漏调节中的作用考虑到ROS在肺血管渗漏中的主要作用， 在核心B强有力的协助下，PMN贩运，具体目标#1（SA #1）将表征ROS的作用， 感应转录因子（HIF-1α/2α，NRF 2），启动子SNP（主要在黑人中）和DNA甲基化 S1 PR 1/S1 PR 3和SELPLG/SELP的遗传/表观遗传调控位点。利用引人注目的初步 数据和关键的核心D支持，SA #2将研究AKT硝化，蛋白质 去泛素化（UCHL 1），Rac GT3激活剂DOCK 1和肌动蛋白结合FA蛋白，板脂蛋白， S1 PR 1和S1 PR 3信令。SA #3将表征S1 PR 1/S1 PR 3对PSGL 1分泌的双重影响 和P-选择素膜从EC韦伯-帕拉德体的移动，对EC细胞骨架的信号传导，和 改变EC屏障反应。利用在临床前大鼠和猪LPS/VILI方面无与伦比的Core C专业知识 模型中，SA #4将评估用S1 PR 3或S1 PR 3/TySIPosome进行装载的治疗效果。 siRNA或UCHL 1激活剂（CD 19 H28 N205 S）的作用，并直接评估UCHL 1竞争性抑制剂的功效。 P-选择素-PSGL-1结合，TSGL-Ig生物制剂（串联P-选择素糖蛋白配体-免疫球蛋白）。 因此，项目4研究将解决对ARDS相关EC增加的新见解的未满足需求 用于靶向S1 P1/3/PSGL 1以恢复肺循环的完整性的新型ARDS疗法。