Lipid mediators and their signaling in ocular surface inflammation and meibomian gland dysfunction

眼表炎症和睑板腺功能障碍中的脂质介质及其信号传导

• 批准号: 10013711
• 负责人: Anat Galor
• 金额: --
• 依托单位: MIAMI VA HEALTH CARE SYSTEM
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-10-01 至 2024-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10013711
• 关键词: Affect; Age; Anti-Inflammatory Agents; Arachidonic Acids; Biological; Biological Assay; Cell Death; Cell Line; Cell Survival; Ceramides; Clinical; Clinical Data; Complex; Dinoprostone; Disease; Docosahexaenoic Acids; Eicosanoids; Eicosapentaenoic Acid; Epithelial Cells; Evaluation; Eye; Female; Film; Foundations; Frequencies; Functional disorder; Goals; Human; In Vitro; Individual; Inflammation; Inflammation Mediators; Inflammatory; Knowledge; Laboratories; Link; Lipids; Measures; Mediating; Modeling; Molecular; Morbidity - disease rate; Nature; Omega-3 Fatty Acids; Omega-6 Fatty Acids; Pain; Pathway interactions; Patients; Phospholipase A2; Pilot Projects; Polyunsaturated Fatty Acids; Production; Prostaglandins; Proteins; Publishing; Quality of life; Regression Analysis; Research; Role; Sample Size; Sampling; Signal Transduction; Signs and Symptoms; Sphingolipids; Sphingomyelinase; Sphingomyelins; Stress; Symptoms; Testing; Time; Veterans; Visual; Work; aqueous; base; ceramide 1-phosphate; evaporation; eye dryness; improved; in vitro Model; in vivo Model; inflammatory marker; inorganic phosphate; lipid mediator; male; meibomian gland; meibomian gland dysfunction; military veteran; molecular targeted therapies; multiplex assay; novel; ocular surface; prospective; sphingosine 1-phosphate; therapeutic development; therapeutic target; treatment optimization

Dry eye (DE) affects 1 in 5 veterans and impacts quality of life. It is a complex disease that manifests with multiple symptoms (e.g. pain, visual disturbance) and signs (e.g. decreased tear production, increased evaporation). Key pathophysiological components of DE are ocular surface inflammation and meibomian gland dysfunction (MGD). Despite its frequency and morbidity, current therapies do not relieve symptoms in all patients. In order to appropriately manage DE, it is necessary to understand mechanisms and specific contributors to disease sub-types. The overall goal of this proposal is to improve such knowledge by studying the role of bioactive sphingolipids (SPL) in MGD and other aspects of DE. Our preliminary and published work suggest SPL composition changes (higher sphingomyelin (SM)/ ceramide (Cer) ratio) in poor quality meibomian gland secretion (meibum); a higher ratio of pro-inflammatory (ω-6) /pro-resolving (ω-3) lipids, and higher levels of prostaglandin E2 in tears from DE patients. Bioactive Cer generated from SM by sphingomyelinases (SMase) and its derivatives, ceramide 1-phosphate and sphingosine 1-phosphate are inflammatory lipids and can induce formation of other inflammatory lipids such as prostaglandins. We detected SMase activity in human tears. In this proposal, we will test the hypothesis that changes in SPL composition in meibum contribute to various signs of DE including ocular surface inflammation, tear film disturbances, and MGD. In Aim 1, we will profile meibum and tear SPL and pro- and anti-inflammatory PUFAs and eicosanoids by LC-MS/MS from 120 cases (poor meibum quality) and 120 controls (normal meibum quality), chosen from our prospectively collected samples from a large veteran population after a comprehensive eye evaluation (mean age 62±10; 467 males, 46 females; 230 white, 272 black). Correlation and regression analysis will be performed with clinical data as dependent variables and lipid mediators as independent variables. In Aim 2, we will determine acitivity of SMAse (acidic and neutral) from 240 Schirmer's strips corresponding to the samples in Aim 1. We will then build models looking at relationships between SMase activity, SPL, and ocular surface inflammation. In Aim 3, we will identify molecular connections between SPL and inflammation in an in vitro model by using human immortalized meibomian gland epithelial cell line (HMGEC) to test the effects of bioactive SPL on mediating inflammation, cell viability, cell death etc. in normal and hyperosmolar conditions. Impact: Our results will identify SPL differences by MGD status, elucidate relationships between bioactive SPL, inflammatory lipid mediators, and SMase activity with ocular surface inflammation and clinical features of MGD and DE sub-types. The knowledge generated will advance the field by determining the relative contributions of different lipids on different manifestations of DE and provide the foundation for developing new molecular targets for therapy.

干眼症（DE）影响5名退伍军人中的1人，并影响生活质量。这是一种复杂的疾病，表现为 多种症状（例如疼痛，视觉灾难）和体征（例如，撕裂产生减少，增加 蒸发）。 DE的关键病理生理组成部分是眼表注射和梅博氏腺 功能障碍（MGD）。尽管其频率和发病率，但目前的疗法并未挽救所有症状 患者。为了适当管理DE，有必要了解机制和特定的机制 疾病子类型的贡献者。该提案的总体目标是通过研究来改善此类知识 生物活性鞘脂（SPL）在MGD和DE的其他方面的作用。我们的初步和出版 工作表明SPL组成的变化（较高的鞘磷脂（SM）/神经酰胺（CER）比率较差） Meibomian腺体分泌（Meibum）；较高的促炎（ω-6） /pro-pro-Crolving（ω-3）脂质的比例较高，并且 DE患者眼泪中的前列腺素E2较高。由SM生成的生物活性CER 鞘磷脂酶（SMASE）及其衍生物，神经酰胺1-磷酸盐和1-磷酸盐是 炎症性脂质，并会诱导其他炎症性脂质（如前列腺素）的形成。我们检测到 在人眼泪中的活性。在此提案中，我们将检验以下假设： Meibum有助于DE的各种迹象，包括眼表面感染，撕裂膜灾难和 MGD。在AIM 1中，我们将介绍Meibum，Tear Spl以及促炎和抗炎的Pufas和类eicosanoids 从120例（较差的Meibum质量）和120个对照（正常的Meibum质量）中，通过LC-MS/MS，从 在全面评估之后，我们从大量退伍军人人口中收集的样品 （平均年龄62±10; 467名男性，46位女性； 230 White，272黑色）。相关和回归分析将是 用临床数据作为因变量和脂质介体作为自变量进行。在AIM 2中，我们 将确定与样品相对应的240个Schirmer条的Smase（酸性和中性）的阳性 在AIM 1中。然后，我们将建立模型，以查看Smase活动，SPL和眼表面之间的关系 在AIM 3中，我们将在体外确定SPL和注射之间的分子连接 通过使用人类永生的Meibomian腺上皮细胞系（HMGEC）测试模型 生物活性SPL在正常和高质量条件下介导感染，细胞活力，细胞死亡等。 影响：我们的结果将通过MGD状态确定SPL的差异，阐明生物活性之间的关系 SPL，炎性脂质介质以及具有眼表面感染的SMASE活性和临床特征 MGD和DE子类型。产生的知识将通过确定相对来推进领域 不同脂质对DE不同表现的贡献，并为开发新的贡献提供了基础 分子治疗靶标。