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: 10293544
• 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/10293544
• 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 (DE) 影响五分之一的退伍军人，并影响生活质量。这是一种复杂的疾病，表现为以下症状。 多种症状（例如疼痛、视力障碍）和体征（例如泪液分泌减少、泪液分泌增多） DE 的关键病理生理学成分是眼表炎症和睑板腺。 尽管其频率和发病率很高，但目前的治疗方法并不能缓解所有症状。 为了正确管理 DE，有必要了解机制和具体情况。 该提案的总体目标是通过研究来提高这些知识。 生物活性鞘脂 (SPL) 在 MGD 和 DE 其他方面的作用。 研究表明，SPL 成分发生变化（鞘磷脂 (SM)/神经酰胺 (Cer) 比例较高），质量较差 睑板腺分泌（睑脂）；促炎 (ω-6)/促消解 (ω-3) 脂质的比例较高，以及 DE 患者的泪液中前列腺素 E2 水平较高，由 SM 产生的生物活性 Cer 产生。 鞘磷脂酶 (SMase) 及其衍生物、神经酰胺 1-磷酸和鞘氨醇 1-磷酸 我们检测到，它可以诱导炎症脂质的形成，例如前列腺素。 人类眼泪中的 SMase 活性在本提案中，我们将测试 SPL 成分发生变化的假设。 睑脂导致 DE 的各种症状，包括眼表炎症、泪膜紊乱和 MGD。在目标 1 中，我们将分析睑脂和泪液 SPL、促炎和抗炎 PUFA 以及类二十烷酸。 通过 LC-MS/MS 对 120 个病例（睑脂质量差）和 120 个对照（睑脂质量正常）进行分析，选自 经过全面的眼部评估后，我们前瞻性地从大量退伍军人中收集了样本 （平均年龄 62±10；467 名男性，46 名女性；230 名白人，272 名黑人）将进行相关和回归分析。 在目标 2 中，我们以临床数据作为因变量和脂质介质作为自变量进行。 将测定与样品相对应的 240 个 Schirmer 试纸条中 SMAse（酸性和中性）的活性 目标 1。然后我们将建立模型来研究 SMase 活性、SPL 和眼表之间的关系 在目标 3 中，我们将在体外鉴定 SPL 与炎症之间的分子联系。 使用人永生化睑板腺上皮细胞系（HMGEC）建立模型来测试 生物活性 SPL 在正常和高渗条件下介导炎症、细胞活力、细胞死亡等。 影响：我们的结果将根据 MGD 状态识别 SPL 差异，阐明生物活性之间的关系 SPL、炎症脂质介质和 SMase 活性与眼表炎症和临床特征的关系 MGD 和 DE 子类型所产生的知识将通过确定相关性来推动该领域的发展。 不同脂质对DE不同表现的贡献，并为开发新的脂质体提供基础 治疗的分子靶点。