Host Vascular Mechanism for Therapeutic Protection against Select Pathogens

项目概要/摘要 许多 NIAID A 类病原体已发展出通过破坏宿主细胞来伤害宿主的机制。 内皮屏障诱导进行性血管渗漏。这会导致肺水肿和呼吸系统疾病 问题、出血热或休克样症状。我们假设改善血管弹性和 完整性可以保护宿主免受多种传染源的侵害。我们将研究关键的作用 使用斑马鱼模型和 R21 中基于内皮细胞的检测来调节内皮完整性 本研究的阶段。使用炭疽致命毒素作为血管渗漏的诱导剂，我们将确定如何 改变宿主血管生成信号可以改善内皮屏障完整性和血管功能。在R33中 在这个项目的阶段，我们将启动一个大规模的化学库筛选，以识别积极的命中和 描述它们的生物学效应。我们将利用当前关于 3 个关键血管生成的知识 因子及其受体 VEGF-A/VEGFR2、血管生成素 1/Tie2 和 ephrinB2/EphB4，以建立可靠的 血管完整性模型。这些模型将用于识别可以改善血管的小分子 恢复力作为潜在的宿主靶向疗法。我们建议在年内完成以下具体目标 R21 阶段：目标 1. 确定血管配体与炭疽致死毒素相比的独特作用 (LT)，使用斑马鱼和内皮细胞模型调节通透性；目标2. 创造条件 使用斑马鱼和血管内皮细胞模型进行小规模化合物筛选 通过使用 NERCE 化学资源实现渗透。将评估这些目标的实现情况 通过 5 个里程碑，使该项目能够进入 R33 阶段。的目标 R33 如下： 目标 3. 筛选化学库中可以改善血管完整性的化合物， R33、Y3-4；目标 4. 评估所选阳性化合物的剂量反应并确定是否 结构改变可以提高血管弹性，同时减少斑马鱼模型中的不利影响，如 以及血液和淋巴管内皮细胞渗漏测定，R33，Y4-5。该项目旨在测试 关于血管保护作为针对病原体攻击的宿主靶向方法的假设。我们设想 R21阶段的积极结果将有助于确定该项目R33阶段的筛选策略。 获得当地化学资源和专业知识将加快这些努力。

PROJECT SUMMARY/ABSTRACT A number of NIAID Category A pathogens has developed mechanisms to injure the host through damaging the endothelial barrier to induce progressive vascular leakage. This leads to pulmonary edema and respiratory problems, hemorrhagic fever or shock-like symptoms. We hypothesize that improving vascular resilience and integrity could protect the host against a number of infectious agents. We will investigate the role of key regulators of endothelial integrity using the zebrafish model and in endothelial cell-based assays in the R21 phase of this study. Using anthrax lethal toxin as an inducer of vascular leakage, we will determine how altering host angiogenic signaling could improve endothelial barrier integrity and vascular function. In the R33 phase of this project, we will launch a large-scale chemical library screen, to identify positive hits and characterize their biological effects. We will take advantage of the current knowledge on 3 key angiogenic factors and their receptors, VEGF-A/VEGFR2, Angiopoietin 1/Tie2 and ephrinB2/EphB4, to establish reliable models for vascular integrity. These models will be used to identify small molecules that can improve vascular resilience as potential host-targeted therapies. We propose the following Specific Aims to be completed in the R21 phase: Aim 1. To establish the distinct roles of vascular ligands in comparison with anthrax lethal toxin (LT), in the regulation of permeability using zebrafish and endothelial cell models; Aim 2. To develop conditions for a small scale chemical compound screening using the zebrafish and endothelial cell models of vascular permeability through the use of NERCE chemical resources. Achievement of these Aims will be evaluated through 5 Milestones that would enable progression to the R33 phase of this project. The Aims for the R33 are as follows: Aim 3. To screen chemical libraries for compounds that can improve vascular integrity, R33, Y3-4; Aim 4. To evaluate the dose-response of selected positive compounds and determine whether structural alterations can improve vascular resilience while reducing adverse effects in the zebrafish model, as well as in blood and lymphatic endothelial cell leakage assays, R33, Y4-5. This project is designed to test a hypothesis on vascular protection as a host-targeted approach to pathogen challenge. We envisioned that positive results in the R21 phase will help define the screening strategy in the R33 phase of this project. Access to local chemical resources and expertise will expedite these efforts.