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

基本信息

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
项目类别：
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

项目摘要

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 大流行极大地凸显了 ARDS 的独特血管内型，其特征是宽内皮细胞 (EC) 通透性、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 FDA 批准的任何有效疗法来阻止或逆转致命性血管渗漏 ARDS/VILI。肺 EC 屏障完整性丧失会导致血管渗漏、肺泡充盈，这是一个关键特征。 ARDS 多器官衰竭和死亡率项目 #4 的研究人员已经证明，sphingosine-1-phos- 多功能脂质介质磷酸酯 (S1P) 及其类似物泰西膦酸酯 (TySIP) 可有效减少血管通过连接 S1PR1（一种在肺中高度表达的 G 蛋白偶联受体）导致渗漏和炎症性肺损伤 S1PR1 连接迅速启动信号级联，通过 Rac 重组 EC 细胞骨架。 GTPase 信号传导可增强连接完整性，并降低血管通透性。 S1PR3 受体诱导 Rho GTPase 信号传导至细胞骨架，以增加肺通透性。此外， S1PR3 被释放到 VILI 或 LPS 暴露小鼠的循环微泡中，作为一种新型 ARDS 双预测 S1PR1 和 S1PR3 启动子活性、表达和下游信号传导的标记物。受具有选择性启动子去甲基化的机械应力的病理生理水平的影响，并且我们将 SELPLG 确定为与 ARDS 风险相关的启动子 SNP。 Blacks，编码 P-选择素糖蛋白配体 1 (PSGL1)，通过 P-选择素 (Selp) 结合至关重要我们令人兴奋的数据表明 S1PR/S1PR 和 S1PR 之间存在相互关联的信号。鉴于 ROS 在肺血管渗漏中的主要作用，PSGL1/SELPLG 在 EC 血管渗漏的调节中。 PMN 贩运，具体目标 #1 (SA #1)，在强大的核心 B 协助下，将表征 ROS 的作用- 传感转录因子（HIF-1α/2α、NRF2）、启动子 SNP（主要在黑人中）和 DNA 甲基化 S1PR1/S1PR3 和 SELPLG/SELP 的遗传/表观遗传调控位点。数据和关键的 Core D 支持，SA #2 将检查 AKT 硝化、蛋白质的新型屏障调节作用去泛素化 (UCHL1)、Rac GTP 酶激活剂 DOCK1 和肌动蛋白结合 FA 蛋白、片脂蛋白 (lamellipodin) S1PR1 和 S1PR3 信号传导 SA #3 将表征 S1PR1/S1PR3 对 PSGL1 分泌的双重影响。和来自 EC Weibel-Palade 小体的 P-选择素膜动员，向 EC 细胞骨架发出信号，以及利用临床前大鼠和猪 LPS/VILI 无与伦比的 Core C 专业知识改变 EC 屏障反应。模型中，SA #4 将评估装载有 S1PR3 的 TySIPosome 的治疗效果 siRNA 或 UCHL1 激活剂 (CD19H28N205S)，并直接评估竞争性抑制剂的功效 P-选择素-PSGL-1 结合，TSGL-Ig 生物制品（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通透性以及针对 S1P1/3/PSGL1 以恢复肺循环完整性的新型 ARDS 疗法。