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：
9903357
负责人：
Haydee E.P. Bazan
金额：
$ 36.5万
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 2022-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9903357
关键词：
Affect Aging Antibodies Axon Biological Assay Brain-Derived Neurotrophic Factor CRISPR/Cas technology Calcium Chemical Burns Clinical Clinical Research Coculture Techniques Cornea Corneal Diseases Corneal Injury Corneal Stroma Corneal Ulcer Dendritic Cells Diabetes Mellitus Diabetic mouse Disease Docosahexaenoic Acids Epithelial Epithelial Cells Epithelium Esthesia Event Fatty Acids Female Flow Cytometry Future Gender Gene Expression Gene Expression Regulation Genes Grant Human Immune Impaired wound healing Impairment Incidence Infection Inflammatory Response Injections Injury Keratoconus Keratoplasty Knockout Mice Knowledge Lead Lecithin Lipids Lymphocyte Mediator of activation protein Membrane Microfluidics Modeling Molecular Molecular Biology Techniques Molecular Mechanisms of Action Multiple Sclerosis Mus Natural regeneration Nerve Nerve Regeneration Neurites Neurons Neuropeptides Neurotrophic Keratitis Omega-3 Fatty Acids Operative Surgical Procedures Outcome PLA2G6 gene Pathology Perforation Phospholipase Play Procedures Production Publications Research Research Project Grants Role Semaphorins Signal Pathway Sjogren&apos s Syndrome Structure of trigeminal ganglion Surface Testing Therapeutic Agents Tissues afferent nerve behavior measurement blink reflexes corneal epithelium corneal regeneration corneal surgery density diabetic effective therapy experimental study eye dryness functional restoration immunoregulation in vivo in vivo Model inflammatory modulation injured innovation lipid mediator macrophage male melting mouse model nerve supply neuroprotectin D1 neurotransmission neurotrophic factor neutrophil novel ophthalmic nerve pigment epithelium-derived factor pigment epithelium-derived factor receptor preservation programs protein expression regenerative response restoration selective expression sex tandem mass spectrometry

项目摘要

Abstract 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, cornea transplant, herpetic infection, chemical burns, keratoconus, multiple sclerosis, Sjogren’s syndrome, aging 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 the w-3 fatty acid docosahexaenoic acid (DHA) or the docosanoid derivative neuroprotectin D1 (NPD1) stimulate nerve regeneration after corneal surgery that damages the stromal nerves. We have recently found that: 1) corneas stimulated with PEDF+DHA also synthesize other docosanoids, one of them identified as resolvin D6 (RvD6); 2) treatment with PEDF results in activation of a calcium independent phospholipase A2ζ (iPLA2ζ); and 3) treatment also stimulates the gene expression in the cornea of neurotrophins and Semaphorin 7A (SEMA7A), which are secreted into tears, and neuropeptides in the trigeminal ganglia (TG) of a mouse model. Our objective will be to define the cascade of molecular events that conduct the corneal nerve regeneration stimulated by PEDF+DHA. Our central hypothesis is that PEDF+DHA, through specific docosanoids, activates selective gene programs and modulates the inflammatory response that in turn, induces the nerve regeneration that leads to preservation of corneal integrity. We will employ: 1) PEDF-receptor (PEDF-R) knockout (KO) mice and in vivo models of corneal injury relevant to the clinical setting; 2) microfluidic chambers for co-culture of TG neurons and corneal epithelial cells to define the molecular mechanism of neurite outgrowth; 3) LC-tandem mass spectrometry lipidomic analysis to identify and quantify the incorporation of DHA in membrane phosphatidylcholine molecular species and DHA-derivatives NPD1, RvD6 and other docosanoids; 4) flow cytometry and our immunostaining assays to determine content of lymphocytes, dendritic cells, macrophages and neutrophils; 5) immunostaining to quantify corneal nerves; 6) behavioral measurements of ocular sensation to assess the functionality of the regenerated nerves; and 7) molecular biology techniques, including gene editing to study the role of the different genes involved in the signaling of nerve regeneration activated by PEDF+DHA and docosanoids. The proposed studies target new molecular mechanisms to understand and treat complications due to corneal nerve damage. Our innovative approach will define agents for neurotrophic keratitis and dry eye after refractive surgery.

抽象的角膜神经的改变导致角膜感觉受损，严重的干眼和对上皮可能导致角膜溃疡，融化和穿孔。这些改变经常发生屈光手术后，角膜移植，疱疹感染，化学烧伤，角膜骨骼，多发性硬化症，多发性硬化症， Sjogren综合征，衰老和糖尿病。尽管有缓解严重的干眼症的治疗方法没有疗法可以补偿神经支配的丧失。该研究项目以我们的发现为基础该色素上皮源性因子（PEDF）加上W-3脂肪酸二十六烯酸（DHA）或二战性衍生物神经保护素D1（NPD1）刺激角膜手术后神经再生损害基质神经。我们最近发现：1）用PEDF+DHA刺激角膜合成其他Docosanoids，其中一个被识别为Resolvin D6（RVD6）； 2）用PEDF治疗导致钙独立磷脂酶A2ζ（IPLA2ζ）的激活； 3）治疗还刺激基因在神经营养蛋白和信号素7a（Sema7a）的角膜中的表达，它们被分泌为泪水，而小鼠模型的三叉神经节（TG）中的神经肽。我们的目标是定义通过PEDF+DHA刺激的角膜神经再生的分子事件。我们的中心假设是，PEDF+DHA通过特定的docosanoids激活选择性基因程序和调节炎症反应，而炎症反应又引起了导致的神经再生保持角膜完整性。我们将采用：1）PEDF-peptor（PEDF-R）敲除（KO）小鼠和与临床环境有关的角膜损伤的体内模型； 2）用于TG共同培养的微流体室神经元和角膜上皮细胞，以定义神经落生的分子机制； 3）LC-Tandem 质谱脂肪分析分析，以识别和量化DHA在膜中的掺入磷脂酰胆碱的分子物种和DHA衍生物NPD1，RVD6和其他二战； 4）流细胞仪和我们的免疫染色测定，以确定淋巴细胞，树突状细胞，巨噬细胞的含量和中性粒细胞； 5）免疫染色以量化角膜神经； 6）眼睛的行为测量感觉评估再生神经的功能； 7）分子生物学技术，包括基因编辑以研究涉及的不同基因在神经再生信号传导中的作用 PEDF+DHA和DOCOSANOIDS。拟议的研究针对新的分子机制来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的药物屈光手术后的角膜炎和干眼。