Conjunctival Goblet Cell Mucin Secretion in Inflammation and Its Resolution

描述（由申请人提供）：尽管15-20%的美国人患有过敏性结膜炎，但目前的治疗仅限于抗组胺药和肥大细胞稳定剂，这些药物通常无效。该提案的长期目标是开发一种新的治疗过敏性结膜炎的方法，这种方法也可以用于治疗肺、皮肤和鼻子的过敏以及其他眼表炎症性疾病，如干眼症。新的治疗方法是一种抗炎和促溶解脂质介质，d系列resolvins与目前处于临床试验中的e系列resolvins相比。结膜杯状细胞过度分泌包括黏液蛋白MUC5AC在内的黏液是变应性结膜炎的症状之一，包括瘙痒、结膜红肿、流泪和化脓。D-，而不是E-，级数解析有两种作用模式。它们阻断组胺和白三烯对杯状细胞分泌的过度刺激，自身激活少量分泌，在过度分泌被重置时保护眼表。e系列不增加分泌。目前的建议将集中在确定促炎介质如何在分子水平上发挥作用，以在培养和体内小鼠过敏性结膜炎模型中对抗促炎介质刺激杯状细胞粘蛋白分泌。研究将集中在以下目标：1。确定D系列和e系列resolvins用于刺激杯状细胞粘液分泌的细胞机制，以及杯状细胞是否是resolvins和其他脂质介质的来源；2. 解开

DESCRIPTION (provided by applicant): Although 15-20% of Americans suffer from allergic conjunctivitis, current treatments are limited to anti-histamines and mast cell stabilizers that ar often ineffective. The long-term goal of the proposal is to develop a new treatment for allergic conjunctivitis that could also be used for allergies of the lung, skin, and nose along with other inflammatory diseases of the ocular surface, such as dry eye. The new treatment is an anti-inflammatory and pro-resolution lipid mediator, the D-series resolvins compared to E-series resolvins currently in a clinical trial. Conjunctival goblet cell over-secretion of mucous includin the mucin MUC5AC is one of the symptoms of allergic conjunctivitis that include itching, conjunctival redness, tearing, and chemosis. D-, but not E-, series resolvins have two modes of action. They block histamine and leukotriene over-stimulation of goblet cell secretion and themselves activate a small amount of secretion to protect the ocular surface while the over-secretion is being reset. E-series do not increase secretion. The current proposal will focus on determining how pro-resolution mediators act at a molecular level to counter-regulate proinflammatory mediator stimulation of goblet cell mucin secretion both in culture and in vivo in a mouse model of allergic conjunctivitis. Research will focus on the following aims: 1. Determine the cellular mechanisms the D- and E-series resolvins use to stimulate goblet cell mucin secretion and if goblet cells are the source of the resolvins and other lipid mediators; 2. Unravel the molecular mechanisms D- and E-series resolvins use to activate protein kinases to interact with histamine and leukotriene receptors to counter-regulate their action; and 3. Ascertain the efficacy of D- compared to E-series resolvins to terminate goblet cell secretion and improve symptoms in a murine model of allergic conjunctivitis. For aim 1 RvD1, aspirin triggered (AT)-RvD1, and RvE1 will be used to stimulate human conjunctival goblet cells in culture. The receptors and cellular signaling pathways activated by the resolvins including phospholipase C and D and PI-3K will be investigated using siRNA, pharmacological inhibitors, and adenovirus constructs. In addition, the production of pro-inflammatory and pro-resolution mediators will be determined by lipidomic measurements on goblet cells in culture. For aim 2, CHO cells transfected with a pro-inflammatory histamine or leukotriene receptor and a pro-resolution receptor and cultured human goblet cells will be used. Protein kinases predicted to phosphorylate the pro-inflammatory mediator receptors will be studied to determine if RvD1, AT-RvD1, and RvE1 activate the protein kinases and counter-regulate the histamine or leukotriene receptor to block its activity and terminate mucin secretion. siRNA, pharmacological inhibitors, and adenovirus constructs will be used for this aim. For aim 3, a mouse model of severe allergic conjunctivitis will be used with ovalbumin as the allergen. Resolvins compared to anti-histamines will be used to determine if they decrease MUC5AC secretion and goblet cell proliferation, ameliorate the clinical symptoms of allergic conjunctivitis, and terminate infiltraton of inflammatory cells.