Vascular stabilization as a broad theraprutic platform for biodefense

许多A、B和C类优先病原体的感染激活先天性免疫系统，触发免疫应答。 使内皮屏障不稳定的细胞因子和其他渗透性因子的过度释放。的 血管完整性的丧失导致非心源性水肿、休克、多器官衰竭和死亡。我们提出 一个新的和广泛的治疗平台，减少发病率和死亡率的基础上，我们确定了一个 内皮受体，Robo 4。Robo 4受体被其配体Slit蛋白激活， 完整性的内皮屏障，并干扰下游信号级联从多个 内皮中的渗透性因子包括TNF-α、白细胞介素和凝血酶。在动物模型中， Robo 4在各种损伤（包括机械损伤、局部缺血和 细胞因子风暴我们最近证明，在两种病原体诱导的急性肺损伤的啮齿动物模型中， 这种方法减少了水肿、休克和死亡。最终，我们的目标是创建一个单一的平台， 治疗多种A、B和C类优先病原体的感染。 目的1：确定Robo 4肽激动剂在病毒性出血热啮齿动物模型中的效力。我们 将使用病毒性出血热模型来检查是否在这些非流感病毒中激活Robo 4， 病原体（沙粒病毒出血热病毒）降低死亡率。这一目标是由一个积极的促进 在细菌内毒素和H5 N1动物模型中已经确定了有效性。 目的2：优化Robo 4肽激动剂。我们将优化重组Slit样肽， 通过连续诱变激活Robo 4信号通路。研究将集中在交付，稳定性， 为将来的GMP/GLP研究做准备所需的生产要求。 目的3：内皮和上皮完整性中的Robo信号传导。我们将决定狭缝机器人4 增强内皮细胞-细胞相互作用，并寻求了解分子机制。我们将使用 这作为研究Slit-Robo 1信号传导是否在上皮-细胞-细胞接触中起类似作用的模型。 该项目符合RMRCE病毒治疗综合研究重点， 五年期将与RP 3.1、3.2、2.3和核心C、D和F直接相互作用。

Infection by many Category A, B, and C Priority Pathogens activates the innate immune system triggering an exuberant release of cytokines and other permeability factors that destabilize the endothelial barrier. The loss of vascular integrity results in non-cardiogenic edema, shock, multi-organ failure and death. We propose a new and broad therapeutic platform for reducing morbidity and mortality based on our identification of an endothelial receptor, Robo4. Activation of the Robo4 receptor by its ligand , Slit proteins, preserves the integrity of the endothelial barrier and interferes with the downstream signaling cascades from multiple permeability factors including TNF-alpha, interleukins, and thrombin in the endothelium. In animal models, Robo4 stabilizes the endothelium following diverse insults including mechanical injury, ischemia, and cytokine storm. We have recently demonstrated that in two rodent models of pathogen induced acute lung injury, this approach reduced edema, shock and death. Ultimately, our goal is to create a single platform for treating infections from a variety of Category A, B and C priority pathogens. AIM 1: Determine efficacy of Robo4 peptide agonist in rodent models of viral hemorrhagic fevers. We will use viral hemorrhagic fever models to examine whether activating Robo4 in these non-influenza viral pathogens (arenavirus hemorrhagic fever virus) reduces mortality. This Aim is facilitated by an active collaborative effort that has already established efficacy in bacterial endotoxin and H5N1 animal models. AIM 2: Optimize a Robo4 peptide agonist. We will optimize recombinant Slit-like peptide capable of activating the Robo4 signaling pathway by serial mutagenesis. Studies will focus on delivery, stability, and production requirements needed to prepare for future GMP/GLP studies. AIM 3: Robo signaling in endothelial and epithelial integrity. We will determine whether Slit-Robo4 enhances endothelial cell-cell interactions, and seek to understand the molecular mechanism. We will use this as a model to investigate whether Slit-Robo1 signaling plays a similar role in epithelial-cell-cell contacts. This project fits within the RMRCE Integrated Research Focus on Viral Therapeutics and over the course of the five year period will interact directly with RP3.1, 3.2, 2.3 and cores C, D, and F.