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：
9902497
负责人：
CHIA-YANG LIU
金额：
$ 53.1万
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 2023-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9902497
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 Epithelium 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.

项目概要/摘要角膜上皮 (CE) 由单层基底细胞和 4-5 个非角膜细胞层组成。角化、复层鳞状上皮细胞形成有效屏障，防止体液流失和 CE主要由密度极高的三叉神经感觉传入神经支配，这对环境损害具有极大的敏感性，否则可以保护更深层的环境通过调节眨眼反应和刺激流泪来保护眼组织免受损伤。 CE 分层和神经支配的破坏会对神经元的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。 CE 分层和神经支配的分子和细胞机制角膜形态发生并在成年期持续维持。探索Shp2介导的Ras丝裂原激活蛋白激酶（Ras-MAPK）途径对于多种细胞活动和行为极其重要，但尚未得到研究我们在 CE 分层和神经支配中努力解决 Shp2 介导的 MAPK 信号传导。通过调节 ΔNp63 表达来控制 CE 分层和神经支配，其三个目标是。建议的：目的1是阐明Shp2→Ras→Mek→ΔNp63→E-cadherin信号通路在发育期间和之后的角膜上皮分层。目标2是阐明CE的Shp2→Ras→Mek→ΔNp63→NGF信号轴在发育期间和之后的 CNV1 神经支配。目标 3 是通过过度表达来挽救其他受损的 CE 分层和神经支配 Shp2cko 小鼠中的 ΔNp63α。完成拟议的目标将描述与 Shp2 介导的机制事件相关 Ras-MAPK 通路微调 ΔNp63，进而调节 E-钙粘蛋白和 NGF，以促进角膜发育和维持过程中 CE 的分层和神经支配这些知识具有巨大的潜力，可以促进基于途径的发现。治疗神经营养性角膜病等角膜疾病的分子靶标。