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：
8887413
负责人：
CHIA-YANG LIU
金额：
$ 42.41万
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 2018-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8887413
关键词：
Ablation Adult Affect Alleles Attenuated Biochemical Breeding Cell Differentiation process Cells Conjunctival Epithelium Data Data Set Development Disease Dominant-Negative Mutation Dry Eye Syndromes Epigenetic Process Epithelial Epithelial Cells Epithelium Equilibrium Exhibits Eyelid structure Film Genes Genetic Goals Goblet Cells Health Human Hyperplasia Keratoconjunctivitis Sicca Knock-out Knockout Mice Knowledge Lead Life Luciferases Maintenance Mediating Metaplasia Molecular Morphogenesis Mouse Strains Mucins Mus Organ Output Pathogenesis Pathway interactions Physiological Platelet Factor 4 Play Publishing Regulation Repression Research Role Signal Pathway Signal Transduction Staging Testing Therapeutic Time Transcription Coactivator 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 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（由适用提供）：结膜杯状细胞是合成和秘密粘蛋白来维持眼表面完整性的主要细胞类型。由于杯状细胞异常，催泪膜中缺乏粘蛋白会导致干眼综合征（DES），并影响数百万人的视力和生命。缺乏关于结膜上皮细胞区分形成杯状细胞的调节机制的知识阻碍了DES治疗方案的发展。我们发现，通过在眼部表面上皮（OSDNMAML1）中，通过有条件的负转录共激活因子策划者（DNMAML1）的有条件表达抑制NOTCH，抑制了小鼠模型中的Goblet细胞分化。与野生型小鼠（OSWT）相比，OSDNMAML1的眼表暴露了结膜上皮增生，异常的脱象和杯状细胞形成受损。此外，OSDNMAML1抑制了Krüppel样转录因子4和5基因（KLF4和KLF5）表达和MUC5/AC合成。相反，在TGFßRII条件敲除（CKO）的整个眼睑基质（CKO）中，结膜上皮被扩展并分化为杯状细胞老鼠。扩展的TGFßRiicko结膜上皮强烈表达的SAM点域含有ETS转录因子（SPDEF），该结构域在多个器官的杯状细胞分化中起着至关重要的作用。我们假设固有的规范缺口和TGFß信号通路及其相互作用在结膜杯细胞分化中起关键作用。我们提出了三个旨在检验这一假设的目标。目标1：阐明锯齿状1/Notch1 N1-ICD/RBP-JκMAML1KLF4/5 MUC5/AC轴对结膜杯状细胞分化至关重要。目标2：阐明TGFßRIISMADS SPDEF MUC5/AC轴在Goblet细胞分化中起作用。 AIM 3：描绘Notch和TGFß路径如何相互作用以提出一个物理输出以进行平衡的杯状细胞分化。