Phosphatidylglycerol as a Therapy for Corneal Injury

磷脂酰甘油治疗角膜损伤

• 批准号: 10179401
• 负责人: Wendy B Bollag
• 金额: $ 37.35万
• 依托单位: AUGUSTA UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-30 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10179401
• 关键词: Adverse effects; Affect; Aftercare; Area; Binding; Biological Assay; CD14 Antigen; CD14 gene; Cell Death; Cell Proliferation; Cells; Chemical Burns; Chronic; Confocal Microscopy; Contact Lenses; Cornea; Corneal Abrasion; Corneal Injury; Data; Diabetes Mellitus; Diabetic mouse; Disease; Dose; Dry Eye Syndromes; Dyes; Energy-Generating Resources; Ensure; Epithelial Cells; Experimental Models; Eye; Generations; Genus Hippocampus; Glycolysis; Heat shock proteins; Histology; Immune; Immune response; Immunologic Monitoring; Impaired healing; Impaired wound healing; Impairment; In Vitro; Individual; Infection; Infiltration; Inflammation; Inflammation Mediators; Inflammatory; Injury; Innate Immune Response; Innate Immune System; Knock-out; Knockout Mice; Lipids; Literature; Measures; Mediating; Membrane Potentials; Mitochondria; Molecular; Monitor; Mus; Neutrophil Infiltration; Operative Surgical Procedures; Oxidative Phosphorylation; Pain; Pathway interactions; Patients; Pattern; Pattern recognition receptor; Phosphatidylglycerols; Phospholipids; Physiological; Process; Production; Publishing; Quality of life; Reactive Oxygen Species; Receptor Activation; Research; Resolution; Respiration; Rodent Model; Role; Safety; Signal Transduction; Skin; Skin wound healing; Small Interfering RNA; Sterility; TLR2 gene; TLR4 gene; Testing; Therapeutic; Thick; Time; Toll-like receptors; Topical application; Vision; Wild Type Mouse; Wound models; alpha-Crystallins; aquaporin 3; base; cell injury; cell motility; corneal epithelial wound healing; corneal epithelium; corneal regeneration; diabetic; effective therapy; experimental study; healing; improved; in vivo; in vivo Model; innovation; instrument; knock-down; macrophage; mitochondrial membrane; mouse model; neutralizing antibody; novel; novel therapeutics; prevent; response; wound; wound healing

Project Summary/Abstract Corneal wounds and abrasions occurring as a result of injury, chemical burns, surgery, contact lens wear and dry eye syndrome are painful and can predispose individuals to infection. Although these wounds tend to heal rapidly, in some cases, such as in diabetes, the healing may be delayed or may even fail; in other cases individuals may continue to show persistent or chronic inflammation despite resolution of the injury or infection. This persistent sterile inflammation can interfere with corneal clarity thereby compromising vision; therefore, treatments to inhibit inflammation while improving corneal wound healing are needed. In exciting novel results we have found that a naturally occurring phospholipid, dioleoylphosphatidylglycerol (DOPG), enhances corneal epithelial wound healing in wild-type mice and in an experimental mouse model of impaired corneal wound healing in vivo. These data suggest the possibility of using DOPG to enhance corneal wound healing therapeutically. Information in the literature also supports a potential role for DOPG in suppressing inflammation. Thus, we have recently demonstrated that DOPG suppresses skin inflammation by inhibiting the activation of pattern recognition receptors, such as toll-like receptor-2 (TLR2) and toll-like receptor-4 (TLR4), in response to damage-associated molecular patterns (DAMPs), endogenous molecules released from injured cells to alert the innate immune system to the presence of danger so that a protective immune response can be mounted. Indeed, a recent article examining an in vivo rodent model of sterile inflammation demonstrated that heat shock protein B4 (HSPB4, also known as crystallin Alpha A) released from corneal keratocytes in response to damaged corneal epithelial cells serves as a DAMP to activate TLR2 on corneal macrophages and induce corneal inflammation. In innovative preliminary studies we have found that DOPG can inhibit macrophage inflammatory mediator production in response to HSPB4, suggesting the likelihood that this lipid will suppress this corneal inflammation. Based on our preliminary results, we hypothesize that DOPG will act not only to accelerate corneal wound healing but also to suppress inflammation by inhibiting TLR2 and/or TLR4 activation. In the research proposed, we will test the idea that DOPG will: (1) dose-dependently, safely and physiologically accelerate corneal epithelial wound healing without adverse effects, at an optimal dose to be determined, in normal and diabetes-impaired corneal wound healing mouse models, and (2) inhibit neutrophil infiltration and inflammation in in vivo mouse models of corneal injury through its ability to inhibit TLR2/4 activation. In a third aim we will also determine the mechanism by which DOPG exerts these effects to stimulate corneal epithelial wound healing and inhibit TLR2/4 activation and inflammation. If our hypothesis proves correct, it would suggest the possibility of developing DOPG as a safe and effective treatment to hasten healing of corneal wounds and to prevent inflammation following injury, infection, ophthalmic surgery or other disorders necessitating corneal epithelial wound healing, thereby improving the quality of life for these patients.

项目概要/摘要 由于受伤、化学烧伤、手术、佩戴隐形眼镜而发生的角膜伤口和擦伤 干眼症会带来痛苦，并且会使人容易感染。虽然这些伤口往往 愈合很快，但在某些情况下，例如糖尿病，愈合可能会延迟，甚至可能失败；在其他情况下 尽管损伤或感染已经消退，但个体可能会继续表现出持续性或慢性炎症。 这种持续的无菌性炎症会干扰角膜清晰度，从而损害视力；所以， 需要治疗来抑制炎症，同时改善角膜伤口愈合。在令人兴奋的新颖结果中 我们发现天然存在的磷脂二油酰磷脂酰甘油 (DOPG) 可以增强角膜 野生型小鼠和角膜伤口受损的实验小鼠模型的上皮伤口愈合 体内愈合。这些数据表明使用 DOPG 增强角膜伤口愈合的可能性 治疗上。文献中的信息也支持 DOPG 在抑制 炎。因此，我们最近证明 DOPG 通过抑制 模式识别受体的激活，例如 Toll 样受体 2 (TLR2) 和 Toll 样受体 4 (TLR4) 对损伤相关分子模式 (DAMP) 的反应，即损伤释放的内源性分子 细胞警告先天免疫系统存在危险，以便保护性免疫反应可以 被安装。事实上，最近一篇研究无菌炎症体内啮齿动物模型的文章表明 角膜细胞释放的热休克蛋白 B4（HSPB4，也称为晶状体蛋白 Alpha A） 对受损角膜上皮细胞的反应充当 DAMP 来激活角膜巨噬细胞上的 TLR2， 诱发角膜炎症。在创新性的初步研究中，我们发现 DOPG 可以抑制 巨噬细胞响应 HSPB4 产生炎症介质，表明这种脂质可能 会抑制这种角膜炎症。根据我们的初步结果，我们假设 DOPG 会采取行动 不仅可以加速角膜伤口愈合，还可以通过抑制 TLR2 和/或 TLR4 来抑制炎症 激活。在提出的研究中，我们将测试 DOPG 的想法：(1) 剂量依赖性、安全性和 生理上加速角膜上皮伤口愈合，且无副作用，最佳剂量为 在正常和糖尿病受损的角膜伤口愈合小鼠模型中确定，并且（2）抑制中性粒细胞 通过抑制 TLR2/4 的能力，在角膜损伤的体内小鼠模型中进行浸润和炎症 激活。在第三个目标中，我们还将确定 DOPG 发挥这些作用的机制 刺激角膜上皮伤口愈合并抑制 TLR2/4 激活和炎症。如果我们的假设 证明是正确的，这表明开发 DOPG 作为一种安全有效的治疗方法的可能性，以加速 愈合角膜伤口并预防受伤、感染、眼科手术或其他原因引起的炎症 需要角膜上皮伤口愈合的疾病，从而改善这些患者的生活质量。