Host Vascular Mechanism for Therapeutic Protection against Select Pathogens

描述（由申请方提供）：许多NIAID A类病原体已经形成了通过破坏内皮屏障来损伤宿主的机制，从而诱导进行性血管渗漏。这会导致肺水肿和呼吸问题，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一些感染因子的侵害。我们将在本研究的R21阶段使用斑马鱼模型和基于内皮细胞的测定来研究内皮完整性的关键调节因子的作用。使用炭疽致死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的完整性和血管功能。在该项目的R33阶段，我们将启动一个大规模的化学库筛选，以识别阳性命中并表征其生物效应。我们将利用目前对3种关键血管生成因子及其受体VEGF-A/VEGFR 2、Angiopoietin 1/Tie 2和ephrinB 2/EphB 4的了解，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弹性的小分子，作为潜在的宿主靶向疗法。我们建议在R21阶段完成以下具体目标：目标1。建立血管配体的不同作用相比，炭疽致死毒素（LT），在调节通透性斑马鱼和内皮细胞模型;目的2。通过使用NERCE化学资源，开发使用斑马鱼和血管通透性内皮细胞模型进行小规模化合物筛选的条件。这些目标的实现将通过5个千年来评估，这将使该项目进入R33阶段。R33的目标如下：目标3。为了筛选化学文库中可以改善血管完整性的化合物，R33，Y3-4;目的4。评价所选阳性化合物的剂量反应，并确定结构改变是否可以改善血管弹性，同时减少斑马鱼模型以及血液和淋巴内皮细胞渗漏试验中的不良反应，R33，Y 4 -5。本项目旨在检验血管保护作为病原体挑战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。利用当地的化学资源和专门知识将加快这些努力。 公共卫生相关性：病原体在扰乱宿主生理学的同时诱导特定效应。虽然宿主免疫系统的激活是重要的一步，但许多感染因子通过损伤内皮细胞来改变心血管功能。当这种情况发生时，患者会出现呼吸问题，因为肺部充满了从血管中泄漏出来的液体。内皮细胞排列在人体的整个血管树中，覆盖约100 m2的表面积，并为病原体作用提供最大的宿主组织。通过了解如何改善血管完整性和识别可以诱导内皮弹性的药物，这些药物可能对许多生物防御相关病原体具有广泛的保护作用。

DESCRIPTION (provided by applicant): 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. PUBLIC HEALTH RELEVANCE: Pathogens induce specific effects while disrupting host physiology. Although activation of the host immune system is an important step, many infectious agents alter cardiovascular function through damage to endothelial cells. When this occurs, the patient suffers from respiratory problems because the lungs are filled with fluid that leaked out o the blood vessels. Endothelial cells line the entire vascular tree in the human body, covering approximately 100 m2 of surface area and providing the largest host tissue for pathogen action. By understanding how to improve vascular integrity and identifying drugs that can induce endothelial resilience, these drugs might be widely protective against a number of biodefense-relevant pathogens.