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.

项目摘要： COVID-19大流行极大地强调了ARDS的独特血管内型，其特征是深层内皮细胞（EC）渗透率，血栓形成和微血管病（1; 2）和重要的未满足需求例如严重缺乏任何有效的FDA批准疗法以停止或反向致死性血管泄漏 ards/vili。肺EC屏障完整性的丧失导致血管泄漏，肺泡洪水，并且是关键特征 ARDS多器官失败和死亡率。项目＃4调查人员已经证明，鞘氨醇1-Phos- Phate（S1P），一种多功能脂质介质及其类似物酸（Tysip），有效减少血管通过连接S1PR1的渗漏和炎症肺损伤，S1PR1是一种高度表达的G蛋白偶联受体肺ECS。 S1PR1连接迅速启动信号级联 GTPase信号传导，增强连接完整性并降低血管渗透性。相反，结扎 S1PR3受体诱导Rho GTPase信号传导至细胞骨架以增加肺部渗透性。此外， S1pr3被释放到循环的微泡中，在vili-或LPS暴露的小鼠中，作为一种新型的Ards Bi- 预测生存的Omarker。 S1PR1和S1PR3启动子活性，表达和下游信号传导受选择性启动子脱甲基化的机械应力的病理生理水平以及与ARDS风险相关的启动子SNP。我们将Selplg确定为一种新型的ARDS敏感性基因黑色编码P-选择蛋白糖蛋白配体1（PSGL1），通过P-选择蛋白（SELP）结合至关重要参与PMN贩运。我们令人兴奋的数据表明S1PR/S1PR和 PSGL1/SELPLG调节EC血管泄漏。鉴于ROS在肺血管泄漏中的主要作用和 PMN贩运，特定的目标＃1（SA＃1），具有强大的核心B援助，将表征ROS-的作用传感转录因子（HIF-1α/2α，NRF2），启动子SNP（黑人中的主要）和DNA甲基化 S1PR1/S1PR3和SELPLG/SELP的遗传/表观遗传调节的位点。利用引人注目的初步数据和关键核心D支持，SA＃2将检查Akt硝化的新型屏障调节作用去泛素化（UCHL1），RAC GTPase激活剂DOCK1和肌动蛋白结合FA蛋白，薄片lamellipodin，In S1PR1和S1PR3信号传导。 SA＃3将表征S1PR1/S1PR3对PSGL1分泌的双重影响以及从EC微花甲状腺，向EC细胞骨架发信号和P-选择素膜动员，并 EC屏障的反应改变了。利用临床前大鼠和猪lps/vili的无与伦比的核心C专业知识型号，SA＃4将评估用任何S1PR3的质体的质体的治疗有效性 siRNA或UCHL1激活剂（CD19H28N205S），并直接评估竞争性抑制剂的有效性 p-选择蛋白-PSGL-1结合，TSGL-Ig生物学（串联P-选择蛋白糖蛋白配体配体 - 免疫球蛋白）。这是项目＃4的研究将解决对EC中与ARDS相关的新见解的未满足的需求渗透性和针对靶向S1P1/3/PSGL1的新型ARDS疗法恢复肺循环的完整性。