权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Role of unfolded protein response and COL8A2 in Fuchs corneal dystrophy

未折叠蛋白反应和 COL8A2 在 Fuchs 角膜营养不良中的作用

基本信息

批准号：
8509695
负责人：
ALBERT S JUN
金额：
$ 37.39万
依托单位：
JOHNS HOPKINS UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-09-01 至 2015-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8509695
关键词：
Affect Age Age-Months Alleles Amino Acids Animals Apoptosis Basement membrane Blindness Caspase Cell Culture Techniques Cell Death Cell Line Cell Shape Cell physiology Cells Cellular Morphology Cellular Stress Cessation of life Chronic Collagen Cornea Corneal Diseases Corneal Endothelium Corneal dystrophy Data Descemet&apos s membrane Disease Endothelial Cells Endothelium Event Exhibits Functional disorder Gene Targeting Genes Genetic Transcription Glutamine Goals Human Hydration status Immunofluorescence Immunologic In Vitro Induction of Apoptosis Inherited Keratoplasty Knock-in Mouse Knowledge Laboratories Leucine Link Lysine Mediating Missense Mutation Modeling Mus Mutant Strains Mice Mutation National Eye Institute Operative Surgical Procedures PAWR protein Pathogenesis Pathologic Pathway interactions Patients Phenotype Physiological Population Positioning Attribute Proteins Research Risk Factors Role Specimen Staging Staining method Stains Stress Surface Testing Translations Tryptophan Tunicamycin Ultraviolet Rays Up-Regulation Work caspase 12 clinically relevant density environmental stressor genetic risk factor improved in vivo insight meter modifiable risk mutant novel protein folding protein misfolding response response marker tool

项目摘要

Fuchs endothelial corneal dystrophy (FECD) is the most common and clinically relevant inherited corneal disease in the US population and the third leading cause of corneal transplants in 2007. FECD results from chronic loss of corneal endothelial cells which line the inner surface of the cornea and maintain physiologic hydration necessary for corneal clarity. In some cases, FECD is caused by missense mutations in the alpha 2 collagen VIII (COL8A2) gene. Very little is known about the pathogenesis of FECD due to the fact that only end stage corneas obtained at corneal transplant are available for study. Limited work indicates that apoptosis is occurring in endothelial cells from advanced FECD corneas, although the mechanisms leading to apoptosis are poorly understood. Thus, animal and cell culture models would be invaluable tools to elucidate early events leading to endothelial cell apoptosis in FECD. Our preliminary work includes: 1) identification of markers for the unfolded protein response (UPR) in the endothelium of FECD patient corneas. Proper protein folding is critical for normal function of the cell. The UPR is a highly conserved, broad ranging cellular response induced by an excess of misfolded proteins. Initially, the UPR functions to relieve stress of unfolded proteins through selective alterations in transcription and translation. If protein folding demand and capacity remain unbalanced, the UPR induces cells to undergo apoptosis. In addition, 2) we have developed two gene targeted knock-in mouse lines with different COL8A2 mutations known to cause FECD in humans. Preliminary results show pathologic changes in the basement membrane of the corneal endothelium which are highly similar to those seen in human FECD corneas. The overall hypothesis of this application is that "corneal endothelial cell expression of COL8A2 mutations activates the unfolded protein response and results in endothelial cell death." The availability of COL8A2 FECD knock-in mice with a relevant pathologic phenotype provides an invaluable opportunity to test this hypothesis as follows: Aim 1 will assess COL8A2 mutant mice for corneal endothelial cell apoptosis and will characterize in greater detail pathologic changes in corneal endothelium and the endothelial basement membrane comparable to human FECD corneas. Aim 2 will assess for UPR activation in corneas and cultured corneal endothelial cell lines from COL8A2 mutant mice. Aim 3 will assess for synergistic effects of UV light (a known inducer of the UPR and environmental stressor for the corneal endothelium) on UPR activation in corneas and cultured corneal endothelial cell lines from COL8A2 mutant mice. This proposal seeks to use COL8A2 FECD mutant mice and corneal endothelial cell lines to establish a novel mechanistic link between the UPR, a major cell stress pathway, and corneal endothelial cell apoptosis resulting from FECD mutations in COL8A2. The establishment of such a link would represent a major advance in the National Eye Institute's goals of improving knowledge and treatment of inherited corneal diseases.

富氏内皮性角膜营养不良（FECD）是最常见且与临床相关的遗传性疾病