Host Vascular Mechanism for Therapeutic Protection against Select Pathogens

项目摘要/摘要 一些NIAID A类病原体已经开发出通过破坏宿主的 内皮屏障导致进行性血管渗漏。这会导致肺水肿和呼吸道疾病。 问题、出血热或休克样症状。我们假设改善血管韧性和 正直可以保护宿主免受多种传染病的侵袭。我们将调查KEY的作用 在R21中使用斑马鱼模型和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改善内皮屏障完整性和血管功能。在R33公路上 在这个项目的阶段，我们将推出一个大规模的化学库筛选，以确定积极的打击和 描述它们的生物学效应。我们将利用现有的关于3个关键血管生成的知识 因子及其受体，血管内皮生长因子-A/血管内皮生长因子2，血管生成素1/Tie2和ewitinB2/EphB4，建立可靠的 血管完整性的模型。这些模型将被用来识别可以改善血管的小分子 复原力作为潜在的宿主靶向疗法。我们提出了以下具体目标，在 R21期：目的1.比较血管配体与炭疽致死毒素的不同作用 (Lt)，在斑马鱼和内皮细胞模型中调节通透性；目的2.开发条件 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化学资源实现渗透性。将评估这些目标的实现情况 通过5个里程碑，将使本项目的R33阶段进展。该计划的目标 R33如下：目的3.筛选可改善血管完整性的化合物的化学文库， R33，Y3-4；目的4.评估选定的阳性化合物的剂量-反应并确定 在斑马鱼模型中，结构改变可以改善血管弹性，同时减少不利影响，因为 以及血液和淋巴管内皮细胞渗漏分析，R33，Y4-5。此项目旨在测试 关于血管保护作为病原体挑战的宿主靶向方法的假说。我们预想到 R21阶段的积极结果将有助于确定该项目R33阶段的筛选战略。 获得当地的化学资源和专门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.