New Insights into the Ocular Surface in Health and Disease

对眼表健康和疾病的新见解

基本信息

批准号：
9330185
负责人：
CHIA-YANG LIU
金额：
$ 42.01万
依托单位：
TRUSTEES OF INDIANA UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-09-01 至 2019-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9330185
关键词：
Ablation Abnormal Cell Adult Affect Alleles Attenuated Biochemical Cell Differentiation process Cells Conjunctival Epithelium Data Data Set Disease Dominant-Negative Mutation Dry Eye Syndromes Epigenetic Process Epithelial Epithelial Cells Epithelium Exhibits Eyelid structure Film Gene Expression Genes Genetic Goals Goblet Cells Health Human Hyperplasia Impairment Keratoconjunctivitis Sicca Knock-out Knockout Mice Knowledge Kruppel-like transcription factors Lead Life LoxP-flanked allele Luciferases Maintenance Mediating Metaplasia Molecular Morphogenesis Mouse Strains Mucins Mus Organ Output Pathogenesis Pathway interactions Phenotype Physiological Platelet Factor 4 Play Publishing Repression Research Role Signal Pathway Signal Transduction Testing Therapeutic Time Transcription Coactivator Transcriptional Regulation Transforming Growth Factor beta Transgenic Mice Treatment Protocols Vision Wild Type Mouse base cell type eye dryness in vivo insight interest mouse model notch protein novel ocular surface postnatal promoter public health relevance receptor therapy development transcription factor

项目摘要

DESCRIPTION (provided by applicant): Conjunctival goblet cells are the major cell type that synthesize and secrete mucins for the maintenance of ocular surface integrity. Lack of mucins in the tear film due to goblet cells abnormality causes dry eye syndrome (DES) and affects millions of people's vision and life. The lack of knowledge regarding the regulatory mechanisms by which conjunctival epithelial cells differentiate to form goblet cells hampers the development of treatment regimens for DES. We found that inhibition of Notch via conditional expression of a dominant negative transcriptional coactivator mastermind-like 1 (dnMAML1) in the ocular surface epithelia (OSdnMAML1) suppressed goblet cell differentiation in mouse model. Compared to the wild-type mouse (OSWt), the ocular surface of the OSdnMAML1 exhibited conjunctival epithelial hyperplasia, aberrant desquamation, and impaired goblet cell formation. Moreover, OSdnMAML1 inhibited Krüppel-like transcriptional factor 4 and 5 genes (Klf4 and Klf5) expression and Muc5/ac synthesis. In contrast, conjunctival epithelium was expanded and differentiated into goblet cells in entire eyelid stroma of the TGFßRII conditional knockout (cKO) mice. The expanded TGFßRIIcKO conjunctival epithelium strongly expressed SAM-pointed domain containing ETS transcription factor (SPDEF), which plays a critical role in goblet cell differentiation in multiple organs. We hypothesize that intrinsic canonical Notch and TGFß signaling pathways and their interaction(s) play pivotal roles in conjunctival goblet cell differentiation. We propose three aims to test this hypothesis. Aim 1: To elucidate that Jagged1/Notch1 N1-ICD/Rbp-jκMAML1 Klf4/5 Muc5/ac axis is critical for the conjunctival goblet cell differentiation. Aim 2: To elucidate that TGFßRII Smads SPDEF Muc5/ac axis has a role in goblet cell differentiation. Aim 3: To delineate how Notch and TGFß pathways interact to come up with a physiological output for balanced goblet cell differentiation.

描述（由申请人提供）：结膜杯状细胞是合成和分泌粘蛋白以维持眼表完整性的主要细胞类型，由于杯状细胞异常导致泪膜中缺乏粘蛋白，导致干眼症（DES）并影响数百万人。对结膜上皮细胞分化形成杯状细胞的调节机制缺乏了解，阻碍了 DES 治疗方案的开发。研究发现，与野生型小鼠 (OSWt) 相比，通过眼表上皮 (OSdnMAML1) 中显性负转录共激活因子 mastermind-like 1 (dnMAML1) 的条件表达来抑制 Notch 可以抑制杯状细胞分化。 OSdnMAML1 表面表现出结膜上皮增生、异常脱屑和杯状细胞形成受损。 OSdnMAML1 抑制 Krüppel 样转录因子 4 和 5 基因（Klf4 和 Klf5）表达和 Muc5/ac 合成，相反，TGFβRII 条件性敲除 (cKO) 的整个眼睑基质中的结膜上皮扩张并分化为杯状细胞。扩展的 TGFβRIIcKO 结膜上皮强烈表达含有 ETS 转录因子 (SPDEF) 的 SAM 指向结构域，该结构域在多个器官的杯状细胞分化中发挥着关键作用。我们提出了三个目标来检验这一假设。目标 1：阐明 Jagged1/Notch1 N1-ICD/Rbp-jκMAML1 Klf4/5 Muc5/ac 轴对于结膜杯状细胞分化至关重要。目标 2：阐明 TGFßRII Smads SPDEF Muc5/ac 轴在杯状细胞分化中的作用。目标 3：描述 Notch 和 TGFβ 通路如何相互作用，产生平衡杯状细胞分化的生理输出。