Lipid Mediators in Corneal Nerve Regeneration

角膜神经再生中的脂质介质

基本信息

批准号：
8827346
负责人：
Haydee E.P. Bazan
金额：
$ 35.28万
依托单位：
LSU HEALTH SCIENCES CENTER
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-04-01 至 2016-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8827346
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 Health 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&apos 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 stem success tandem mass spectrometry

项目摘要

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会导致角膜感觉受损，严重的干眼和对上皮的损害，这可能导致角膜溃疡，融化和穿孔。这些改变经常发生在折射手术，角膜移植，疱疹感染，化学烧伤，角膜结构，多发性硬化症，Sjogren综合征和糖尿病的糖尿病之后。尽管有治疗方法可以减轻严重的干眼症，但没有疗法可以补偿神经损失。这项研究项目基于我们的发现，即色素上皮衍生因子（PEDF）加上二十二碳六烯酸（DHA）或二战性衍生物神经蛋白神经蛋白D1（NPD1）会在角膜手术后引起神经再生的神经再生。现在，我们专注于定义PEDF加上DHA或NPD1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体单纯疱疹病毒类型1（HSV-1）的动物模型中这些介体的生物活性性，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通过介导的机制作用，该机制激活胞质磷脂酶A2/细胞外调节激酶（CPLA2/ERK1/2）信号传导以释放DHA，以释放NPD1的合成，从而刺激神经生长因子（NGF）在IN中的表达。转弯，调节角膜神经再生。除了角膜手术外，HSV-1感染会导致敏感性失去，干眼和角膜病变，可能导致基质和上皮损伤。我们建议检验以下假设：NPD1诱导敏感性恢复并再生疱疹病毒感染后的角膜神经。我们将采用：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一级和细胞系培养和共同培养，以发现神经突生长的分子机制； 3）LC-TANDEM质谱脂质分析分析以表征和量化DHA及其衍生物NPD1； 4）气体估计法评估神经再生后不同刺激方式的角膜敏感性； 5）分子生物学技术和我们的免疫染色方法，以量化上皮，基质和基质角膜神经。拟议的研究针对新机制，以理解和治疗因角膜神经损伤而引起的并发症。我们的创新方法将定义神经营养性角膜炎的潜在药物和屈光手术和HSV-1感染后的干眼症。