Targeting the Endothelium in Sepsis

描述(申请人提供)：血管内皮细胞对局部感染的反应是通过削弱屏障功能和促进凝血来传递体液免疫效应物和遏制微生物的传播。在全身感染中，这些保护性反应变得不适应，导致弥漫性血管渗漏和弥漫性血管内凝血。我们假设脓毒症内皮细胞驱动关键疾病的表现，我们寻求创新的方法来靶向和测量这一复杂的过程。目的1将评估两个主要的内皮细胞表面受体PAR1和Tie2的作用，它们的激活可以促进脓毒症时的炎症、血栓形成和屏障功能障碍。我们已经确定了针对PAR1和Tie2的新型类药物分子，并在面对败血症刺激时协同作用，实现内皮细胞的静止。这一目标将询问PAR1和Tie2如何在内皮细胞上合作，以改善脓毒症的临床相关结果。目标2将阐述内皮如何在败血症微血栓形成中起作用。除了评估PAR1和Tie2，这一目标将通过利用遗传小鼠模型中的活体显微镜和新的基于细胞的方法相结合，首次系统描述导致脓毒症早期微血栓形成和晚期凝血障碍的内皮细胞紊乱。目的3将寻找新的非侵入性方法来跟踪脓毒症时血管内皮细胞的动态反应。我们开发了适合高通量分析的定量、实时、人性化的微血管屏障功能障碍和血管内皮细胞血栓形成潜能的分析方法。应用同类中最大的ICU队列之一的血浆，我们将测试这些分析相对于临床指标和传统生物标记物的预测性能。我们还将探索这些检测方法的潜力，以衡量新的候选疗法的有效性。该应用程序将具有互补专业知识、独特资源以及工作效率和协作记录的调查团队团结在一起。我们将追求高度平行的目标，旨在通过定义基本的疾病机制，测试创新的治疗策略，并开发专注于内皮的强大的新诊断工具，在脓毒症中产生突出的影响。

DESCRIPTION (provided by applicant): The vascular endothelium responds to localized infection by weakening barrier function and promoting coagulation to deliver humoral effectors of immunity and contain the spread of microbes. In systemic infection, these same protective responses become maladaptive, leading to diffuse vascular leakage and disseminated intravascular coagulation. We hypothesize that the septic endothelium drives critical disease manifestations, and we seek innovative ways to target and measure this complex process. Aim 1 will evaluate the roles of two major endothelial cell-surface receptors, PAR1 and Tie2, whose activation can promote inflammation, thrombosis, and barrier dysfunction during sepsis. We have identified novel drug-like molecules that target PAR1 and Tie2 and synergize to achieve endothelial quiescence in the face of septic stimuli. This aim will ask how PAR1 and Tie2 collaborate at the endothelium to ameliorate clinically relevant outcomes of sepsis. Aim 2 will address how the endothelium contributes to septic microthromboses. In addition to evaluating PAR1 and Tie2, this aim will provide a first-in-kind systematic description of the endothelial derangements that drive the early microthromboses and late coagulopathy of sepsis by utilizing a combination of intravital microscopy in genetic mouse models and novel cell-based approaches. Aim 3 will seek new non-invasive ways to follow the dynamic responses of the endothelium to sepsis. We have developed quantitative, real-time, humanized assays of microvascular barrier dysfunction and endothelial prothrombotic potential suitable for high-throughput analysis. Applying plasmas from one of the largest ICU cohorts of its kind, we will test the predictive performance of these assays against clinical metrics and conventional biomarkers. We will also explore the potential of these assays to gauge the efficacy of new candidate therapies. This application unites a team of investigators with complementary expertise, unique resources, and a track record of productivity and collaboration. We will pursue highly parallel aims designed to generate outstanding impact in sepsis by defining fundamental disease mechanisms, testing innovative therapeutic strategies, and developing robust new diagnostic tools focused on the endothelium.