Rhinovirus accelerates wound repair via surface signaling EGFR activation

描述（由申请人提供）：气道上皮的表面代表了一个“战场”，宿主在这里拦截来自病原体的信号并激活先天防御以对抗感染。创伤修复是慢性气道疾病中气道上皮细胞对损伤做出反应的基本功能。鼻病毒（RV）感染是导致这些疾病中的几种恶化的原因。我们假设气道上皮细胞对低浓度RV有反应，以加速伤口修复。此外，我们预测高浓度的RV对气道上皮细胞有毒性并抑制伤口修复。由于表皮生长因子受体（EGFR）参与伤口修复，我们假设RV通过激活EGFR的表面信号通路加速伤口修复。我们的长期目标是研究病毒与气道上皮的相互作用，以进一步了解健康和疾病中的先天免疫。在短期内，我们将集中于三个具体目标。(1)我们将研究RV感染对气道上皮损伤修复的浓度依赖性影响，并在损伤的上皮中定位病毒感染。(2)我们将研究RV加速伤口修复中激活EGFR的表面信号通路。(3)最后，我们将检查RV感染所需的分子是否参与这种表面信号级联反应。在初步实验中，RV加速了划痕伤口模型中的伤口修复，这一效果在较高的病毒浓度下没有观察到。为了确定增加病毒浓度的影响，我们将测量伤口修复率和细胞坏死标志物。为了定位RV感染，我们将使用免疫组织化学和荧光显微镜。在初步实验中，EGFR酪氨酸激酶选择性抑制剂阻止RV加速伤口修复，涉及EGFR激活。我们计划使用中和抗体、金属蛋白酶和氧化酶抑制剂、ROS清除剂和siRNA来评估激活EGFR的表面信号级联。此外，我们将测量EGFR配体和ROS的释放。为了评估ICAM-1、神经酰胺和PKC在RV加速伤口修复中的作用，我们将使用抗体来防止病毒附着，并使用神经酰胺和PKC抑制剂。总之，我们认为这种表面信号级联涉及：RV 16-> ICAM-1 ->神经酰胺-PKC-> Duox 1-> ROS-”TACE-”TGF-α-> EGFR磷酸化。RV感染具有临床重要性，因为该病毒是普通感冒的主要原因，并导致哮喘和COPD恶化。由于RV的治疗是不可用的，研究病毒-上皮相互作用可能会导致抗病毒治疗的新靶点，并可能增加我们对气道防御损伤和感染的理解。

DESCRIPTION (provided by applicant): The surface of the airway epithelium represents a "battleground" where the host intercepts signals from pathogens and activates innate defenses to combat infection. Wound repair is an essential function of the airway epithelium in response to injury in chronic airway diseases. Rhinovirus (RV) infection is responsible for exacerbations of several of these diseases. We hypothesize that the airway epithelium responds to low concentrations of RV to accelerate wound repair. Further, we predict that high concentrations of RV are toxic to airway epithelial cells and inhibit wound repair. Because the epidermal growth factor receptor (EGFR) is involved in wound repair, we hypothesize that RV accelerates wound repair via a surface signaling pathway activating EGFR. Our long term goals are to investigate viral interactions with the airway epithelium to further our understanding of innate immunity in health and disease. In the short term we will focus on three specific aims. (1) We will study the concentration-dependent effects of RV infection on airway epithelial wound repair and localize viral infection in the wounded epithelium. (2) We will investigate the surface signaling pathway in RV-accelerated wound repair that activates EGFR. (3) Finally, we will examine whether molecules required for RV infection are involved in this surface signaling cascade. In preliminary experiments, RV accelerated wound repair in a scratch wound model, an effect that was not seen with a higher viral concentration. To determine the effects of increasing concentrations of the virus, we will measure the rate of wound repair and markers of cell necrosis. To localize RV infection we will use immunohistochemistry and fluorescent microscopy. In preliminary experiments an inhibitor selective for EGFR tyrosine kinase prevented RV-accelerated wound repair, implicating EGFR activation. We plan to evaluate the surface signaling cascade activating EGFR, using neutralizing antibodies, metalloprotease and oxidase inhibitors, ROS scavengers, and siRNA. In addition, we will measure the release of EGFR ligands and ROS. To evaluate the role of ICAM-1, ceramide and PKC in RV-accelerated wound repair, we will use an antibody to prevent viral attachment, and ceramide and PKC inhibitors. In summary, we suggest that this surface signaling cascade involves: RV 16-> ICAM-1 -> ceramide-PKC-> Duox1-> ROS-" TACE-" TGF- a-> EGFR phosphorylation. RV infection has clinical importance because the virus is the predominant cause of the common cold and causes exacerbations of asthma and COPD. Because therapies for RV are not available, investigating viral-epithelial interactions may lead to novel targets for anti-viral therapies and may increase our understanding of airway defenses against injury and infection.