Molecular mechanism of corneal epithelial stratification and innervation

角膜上皮分层和神经支配的分子机制

基本信息

批准号：
10376207
负责人：
CHIA-YANG LIU
金额：
$ 6.36万
依托单位：
TRUSTEES OF INDIANA UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-04-01 至 2022-07-01
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10376207
关键词：
ARHGEF5 gene Adult Axon Basal Cell Behavior Biological Assay Biological Process Blindness Blinking Cell Line Cell Nucleus Cells Coculture Techniques Complex Cornea Corneal Diseases Cytomegalovirus Data Degenerative Disorder Development Disease Down-Regulation E-Cadherin Epithelial Epithelial Cells Esthesia Event Genes Histopathology Homeostasis Human Impairment In Vitro Keratopathy Knock-out Knowledge Lacrimation Lead Liquid substance Luciferases MAP Kinase Gene MAPK Signaling Pathway Pathway MEKs Maintenance Measures Mediating Mitogen-Activated Protein Kinases Molecular Molecular Target Monitor Morphogenesis Mouse Strains Mus Nerve Nerve Growth Factors Neurites Pathway interactions Play Proteins RAS genes Regulation Role Sensory Signal Pathway Signal Transduction Small Interfering RNA Squamous Epithelium Stratification Stratified Epithelium Stratified Squamous Epithelium Stratum Basale Structure of trigeminal ganglion Telomerase Thinness Tissues Transfection Transgenic Mice Trigeminal System Trigeminal nerve structure Ubiquitin Western Blotting base cell growth chromatin immunoprecipitation conditional knockout corneal epithelium density dosage experimental study extracellular gain of function ganglion cell knock-down migration multicatalytic endopeptidase complex mutant nerve supply new therapeutic target novel ocular surface overexpression pathogen plasmid DNA postnatal promoter response therapeutic target

项目摘要

PROJECT SUMMARY/ABSTRACT Corneal epithelium (CE) is composed of a single layer of basal cells and 4-5 cell layers of non- keratinized, stratified squamous epithelial cells to form an effective barrier against fluid loss and pathogen. CE is mainly innervated by trigeminal sensory afferents with extremely high density, which offers enormous sensitivity to the environmental insults for protecting otherwise deeper ocular tissues from damage by modulating the blink response and stimulating lacrimation. Any disruption of CE stratification and innervation can have deleterious effects on the integrity of the cornea and lead to neurotrophic keratopathy and corneal blindness. Little is known regarding the molecular and cellular mechanisms by which CE stratification and innervation is achieved during corneal morphogenesis and continuously sustained in the adulthood. In this proposal, we attempt to explore Shp2-mediated Ras–mitogen-activated protein kinase (Ras-MAPK) pathway which is extremely important for wide variety of cellular activities and behaviors, but it has not been studied in the CE stratification and innervation. We hypothesize that Shp2-mediated MAPK signaling controls CE stratification and innervation via regulating ∆Np63 expression. Three aims are proposed: Aim 1 is to elucidate the role of Shp2→Ras→Mek→∆Np63→E-cadherin signaling pathway in corneal epithelial stratification during and following development. Aim 2 is to elucidate the role of Shp2→Ras→Mek→∆Np63→NGF signaling axis of the CE on the CNV1 innervations during and following development. Aim 3 is to rescue otherwise impaired CE stratification and innervation by overexpression of ∆Np63α in Shp2cko mice. Completion of proposed objectives will delineate mechanistic event related to the Shp2-mediated Ras-MAPK pathway to fine-tune ∆Np63, which in turn regulate E-cadherin and NGF, to faciliate stratification and innervation of CE during development and in the maintenance of corneal homeostasis. This knowledge has great potential to lead to the discovery of pathway-based molecular target to treat corneal diseases such as neurotrophic keratopathy.

项目摘要/摘要角膜上皮（CE）由单层基本细胞和4-5个细胞层组成角膜化的，分层的鳞状上皮细胞，形成有效的屏障，以防止流体损失和病原。 CE主要由非常高密度的三叉神经感官传入支配，这对环境侮辱具有巨大的敏感性，以保护原本更深层次通过调节眨眼反应和刺激富集来造成损伤的眼组织。任何 CE分层和神经神经的破坏会对完整性产生有害影响角膜并导致神经营养性角膜病和角膜失明。关于通过在CE分层和神经上实现CE分层和细胞的分子和细胞机制角膜形态发生并在成年期间连续维持。在此提案中，我们尝试探索SHP2介导的RAS- MIMET基因激活的蛋白激酶（RAS-MAPK）途径对于各种细胞活动和行为极为重要，但尚未研究在CE分层和神经上。我们假设SHP2介导的MAPK信号传导通过调节ΔNP63表达控制CE分层和神经。三个目标是建议的： AIM 1是阐明SHP2→RAS→MEK→∆NP63→E-Cadherin信号通路的作用角膜上皮分层在发育过程中和之后。 AIM 2是阐明SHP2→RAS→MEK→∆NP63→NGF信号轴的作用 CNV1在发育过程中和之后的神经。 AIM 3是通过过表达来营救其他受损的CE分层和神经支配 SHP2CKO小鼠中的∆NP63α。提出的目标的完成将描述与SHP2介导的机械事件 RAS-MAPK通往微调∆NP63的途径又调节E-钙粘蛋白和NGF，以促进 CE在开发和维持角膜过程中的分层和神经支配稳态。这些知识具有巨大的潜力，可以发现基于途径的发现治疗角膜疾病的分子靶标，例如神经营养性角膜病。