Lipid Mediators in Corneal Nerve Regeneration

角膜神经再生中的脂质介质

• 批准号: 8500660
• 负责人: Haydee E.P. Bazan
• 金额: $ 36万
• 依托单位: LSU HEALTH SCIENCES CENTER
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-04-01 至 2017-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8500660
• 关键词: Affect; Angiogenesis Inhibitors; Animal Model; Cell Line; Chemical Burns; Clinical; Coculture Techniques; Cornea; Corneal Injury; Corneal Ulcer; Cytosolic Phospholipase A2; Diabetes Mellitus; Disease; Docosahexaenoic Acids; Epithelial; Epithelial Cells; Epithelium; Esthesia; Fatty Acids; Fibroblasts; Free Nerve Ending; Gases; Grant; Herpesvirus 1; Incidence; Infection; Inflammation; Keratitis; Keratoconus; Keratoplasty; Lead; Length; Lesion; Lipids; MAPK3 gene; Mediating; Mediator of activation protein; Membrane Lipids; Methods; Modality; Modeling; Molecular; Molecular Biology Techniques; Multiple Sclerosis; Natural regeneration; Nerve; Nerve Fibers; Nerve Growth Factors; Nerve Regeneration; Neurites; Neurons; Neurotrophic Tyrosine Kinase Receptor Type 1; Omega-3 Fatty Acids; Operative Surgical Procedures; Oryctolagus cuniculus; Pathway interactions; Perforation; Phosphorylation; Phosphotransferases; Pigments; Play; Procedures; Progress Reports; Property; Receptor Signaling; Recovery; Research; Research Project Grants; Role; Serpins; Signal Pathway; Signal Transduction; Sjogren' s Syndrome; Stimulus; Stromal Cells; Structure of trigeminal ganglion; Testing; Therapeutic Agents; Time; Viral; Virus Diseases; Wound Healing; blink reflexes; corneal epithelium; corneal surgery; effective therapy; extracellular; eye dryness; improved; in vivo; in vivo Model; innovation; insight; lipid mediator; melting; neovascularization; nerve injury; nerve supply; neuroprotectin D1; neurotrophic factor; novel; ocular surface; pigment epithelium-derived factor; prevent; public health relevance; stem; success; tandem mass spectrometry

DESCRIPTION (provided by applicant): Alterations in corneal innervation result in impaired corneal sensation, severe dry eye and damage to the epithelium that may in turn lead to corneal ulcers, melting and perforation. These alterations frequently occur after refractive surgery, corneal transplant, herpetic infection, chemical burns, keratoconus, multiple sclerosis, Sjogren's syndrome and diabetes mellitus. Although there are treatments to alleviate severe dry eye, there are no therapies to compensate for the loss of innervation. This research project builds upon our finding that pigment epithelium-derived factor (PEDF) plus docosahexaenoic acid (DHA) or the docosanoid derivative neuroprotectin D1 (NPD1) induces nerve regeneration after corneal surgery that damages stromal nerves. We now focus on defining the molecular mechanism by which PEDF plus DHA or NPD1 regenerates corneal nerves after experimental refractive surgery. We additionally investigate the bioactivity of these mediators in an animal model of herpes simplex virus type-1 (HSV-1), focusing on compromised sensitivity, nerve regeneration and inflammation. Our central hypothesis is that PEDF acts through a mediated mechanism that activates cytosolic phospholipase A2/extracellular-regulated kinase (cPLA2/ERK1/2) signaling to release DHA for the synthesis of NPD1, which stimulates nerve growth factor (NGF) expression that, in turn, modulates corneal nerve regeneration. In addition to corneal surgery, HSV-1 infections cause lost sensitivity, dry eye and corneal lesions that could lead to stromal and epithelial damage. We propose to test the hypothesis that NPD1 induces recovery of sensitivity and regenerates corneal nerves after herpes viral infection. We will employ: 1) in vivo models of refractive surgery and HSV-1 infection relevant to the clinical setting; 2) primary and cell line cultures and co-cultures of trigeminal ganglion neurons with corneal epithelial cells and stromal fibroblasts to uncover the molecular mechanisms of neurite outgrowth; 3) LC-tandem mass- spectrometry lipidomic analysis to characterize and quantify DHA and its derivative, NPD1; 4) gas esthesiometry to assess corneal sensitivity to different modalities of stimulus after nerve regeneration; and 5) molecular biology techniques and our immunostaining method to quantify the epithelial, sub-basal and stromal corneal nerves. The proposed studies target new mechanisms to understand and treat complications due to corneal nerve damage. Our innovative approach will define potential agents for neurotrophic keratitis and dry eye after refractive surgery and HSV-1 infection.

描述（由申请人提供）：角膜神经支配的改变导致角膜感觉受损、严重干眼症和上皮损伤，进而可能导致角膜溃疡、熔化和穿孔。这些改变经常发生在屈光手术、角膜移植、疱疹感染、化学烧伤、圆锥角膜、多发性硬化、干燥综合征和糖尿病之后。虽然有缓解严重干眼症的治疗方法，但没有治疗方法来补偿神经支配的损失。该研究项目建立在我们的发现之上，即色素上皮衍生因子（PEDF）加二十二碳六烯酸（DHA）或二十二碳烷衍生物神经保护素D1（NPD 1）可诱导损伤基质神经的角膜手术后的神经再生。我们现在专注于定义PEDF加DHA或NPD 1在实验性屈光手术后再生角膜神经的分子机制。我们还研究了这些介质在单纯疱疹病毒1型（HSV-1）动物模型中的生物活性，重点是受损的敏感性，神经再生和炎症。我们的中心假设是PEDF通过介导的机制起作用，该机制激活胞质磷脂酶A2/细胞外调节激酶（cPLA 2/ERK 1/2）信号传导，释放DHA用于NPD 1的合成，NPD 1刺激神经生长因子（NGF）表达，进而调节角膜神经再生。除角膜手术外，HSV-1感染还会导致敏感性丧失、干眼和角膜病变，从而导致基质和上皮损伤。我们建议测试的假设，NPD 1诱导恢复敏感性和再生角膜神经疱疹病毒感染后。我们将雇用：1）与临床环境相关的屈光手术和HSV-1感染的体内模型; 2）三叉神经节神经元与角膜上皮细胞和基质成纤维细胞的原代和细胞系培养物和共培养物，以揭示神经突生长的分子机制; 3）LC-串联质谱脂质组学分析，以表征和定量DHA及其衍生物NPD 1; 4）气体感觉测量法，以评估神经再生后角膜对不同形式的刺激的敏感性;以及5）分子生物学技术和我们的免疫染色方法，以量化上皮、基底下和基质角膜神经。拟议的研究旨在了解和治疗角膜神经损伤引起的并发症的新机制。我们的创新方法将确定潜在的代理人神经营养性角膜炎和干眼症后屈光手术和HSV-1感染。