Conjunctival Goblet Cell Mucin Secretion in Inflammation and Its Resolution

项目总结/摘要 作为对环境的反应，结膜杯状细胞的功能是维持健康的泪膜和眼内分泌。 表面，并防止炎症性疾病。杯状细胞分泌粘蛋白MUC 5AC，其保护性地 眼睛表面。杯状细胞通过积极地保持泪膜的最佳粘液层， 在正常的健康水平。然而，在疾病中，粘蛋白可以过度产生，如过敏性结膜炎。我们 该实验室致力于杯状细胞粘蛋白生产在健康和炎症性疾病中的作用 过敏性结膜炎专门的前分解分子（SPM）是由ω-3脂肪酸产生的 例如DHA。有一个复杂的和严格管制的生物学使用前分辨率分子来控制 杯状细胞分泌粘蛋白，维持眼表稳态。在疾病中， 这可能导致SPM解决的病理生理学的粘蛋白的失调/过度产生。我们的整体 目的是研究结膜是如何产生SPM的，并以性别依赖的方式在一个特定的年龄段起作用。 在健康和疾病中，结膜杯状细胞在分子水平上调节粘蛋白的分泌。我们将专注于 存在于泪液和结膜中的消退素Ds RvD 1 -6。RvD 1和RvD 2刺激杯状细胞 功能在具体目标1中，我们将关注健康并确定哪些RvD家族成员（RvD 3 -6） 增加细胞内[Ca ~（2+）]（[Ca ~（2+）]i），升高cAMP，并刺激培养的结膜粘液分泌 杯状细胞对于这一目标和所有三个目标，我们将评估是否存在反应的性别依赖性差异。在 具体目标2我们将研究在健康状态下RvD 1是否使用具有GPR 32（受体）/DHA的自分泌回路 （前体）/RvD 1轴来控制其生物合成。在具体目标3中，我们将研究疾病和过敏 结膜炎询问哪些RvD家族成员（RvD 2 -6）对抗促炎介质刺激 杯状细胞内[Ca ~（2+）]i增加，分泌增加，促进AED消退。我们将使用人类 培养的结膜杯状细胞，两种SPM受体敲除小鼠，和过敏性小鼠模型 眼疾。细胞内[Ca 2 +]、cAMP、MUC 5AC分泌、DHA释放和RvD 1生物合成将被 测定了在动物模型中，临床症状、MUC 5AC分泌和白细胞运输将被 测定

Project Summary/Abstract In response to the environment, the conjunctival goblet cells function to maintain a healthy tear film and ocular surface, and to prevent inflammatory diseases. Goblet cells secrete the mucin MUC5AC that is protective to the ocular surface. Goblet cells maintain an optimal mucous layer of the tear film by actively keeping it at normal levels in health. In disease, however, mucins can be over produced as in allergic conjunctivitis. Our laboratory concentrates on the role of goblet cell mucin production in health and in the inflammatory disease allergic conjunctivitis. The specialized pro-resolving molecules (SPMs) are produced from omega-3 fatty acids such as DHA. There is a complex and tightly regulated biology of using pro-resolution molecules to control goblet cell mucin secretion and maintain ocular surface homeostasis. In disease there is an allergy-mediated dysregulation/overproduction of mucins that contribute to the pathophysiology that SPMs resolve. Our overall goal is to investigate how SPMs are produced by the conjunctiva and act in a sex-dependent manner at a molecular level in conjunctival goblet cells to regulate mucin secretion in health and disease. We will focus on the resolvin Ds RvD1-6 that are present in tears and conjunctiva. RvD1 and RvD2 stimulate goblet cell function. In Specific Aim 1 we will focus on health and determine which RvD family members (RvD3-6) increase the intracellular [Ca2+] ([Ca2+]i), elevate cAMP, and stimulate mucin secretion in cultured conjunctival goblet cells. For this and all three aims we will evaluate if there a sex-dependent difference in response. In Specific Aim 2 we will investigate if in health RvD1 uses an autocrine circuit with a GPR32 (receptor) /DHA (precursor)/RvD1 axis to control its biosynthesis. In Specific Aim 3 we will study disease and in allergic conjunctivitis interrogate which RvD family members (RvD2-6) counter pro-inflammatory mediator stimulated goblet cell increase in [Ca2+]i and secretion, and promote resolution of AED in vivo. We will use human cultured conjunctival goblet cells, two types of SPM receptor knock out mice, and a mouse model of allergic eye disease. Intracellular [Ca2+], cAMP, MUC5AC secretion, DHA release, and RvD1 biosynthesis will be measured. In the animal model clinical symptoms, MUC5AC secretion, and leukocyte trafficking will be determined.