Determining Molecular Mechanisms of Human Glaucoma Genes

确定人类青光眼基因的分子机制

基本信息

批准号：
10612930
负责人：
Kayarat Saidas Nair
金额：
$ 38.13万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-05-01 至 2026-03-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10612930
关键词：
Affect Alleles Aqueous Humor Binding Sites Biological Assay Biological Process Biology Blindness Cell Adhesion Cell Culture Techniques Cells ChIP-seq Clustered Regularly Interspaced Short Palindromic Repeats Complex Consensus Cytoskeletal Modeling DNA Binding Defect Development Dexamethasone Disease Disease Progression Drainage procedure Enhancers Exhibits Extracellular Matrix Extracellular Matrix Proteins FOXC1 gene Functional disorder Gene Expression Gene Expression Regulation Genes Genetic Genetic Transcription Genomic approach Genomics Glaucoma Goals Heterozygote Homeostasis Human Human Genetics Impairment Introns Maintenance Mediating Modeling Molecular Mus Nucleic Acid Regulatory Sequences Pathogenesis Pathway interactions Persons Physiologic Intraocular Pressure Play Prevalence Primary Open Angle Glaucoma Promoter Regions Public Health Regulation Regulator Genes Reporter Resistance Risk Factors Role Structure of sinus venosus of sclera Study models Susceptibility Gene System Testing Tissues Trabecular meshwork structure Transforming Growth Factor beta Untranslated RNA Variant functional genomics genetic approach high intraocular pressure in vitro Assay insight lentiviral-mediated mouse model optic nerve disorder overexpression prevent programs targeted treatment therapeutic target transcription factor transcription regulatory network transcriptome sequencing

项目摘要

ABSTRACT Glaucoma, a major cause of blindness worldwide, is a significant public health concern. In the U.S., it affects over 2.7 million people and its prevalence will rise to 7.3 million by 2050. Targeted therapies are needed to prevent glaucoma or slow its progression. A major risk factor is high intraocular pressure (IOP), typically due to impaired aqueous humor (AqH) outflow. However, the genes and pathways involved are poorly understood. We have identified GLIS1, encoding the transcription factor GLIS1, as a susceptibility gene for primary open- angle glaucoma (POAG) and showed that Glis1–/– mice have pathophysiological hallmarks of glaucoma. We also found that Glis1 is predominantly expressed in the trabecular meshwork (TM), a key component of the ocular drainage tissue regulating AqH outflow, and that Glis1–/– mice exhibit progressive TM degeneration, leading to high IOP, and glaucomatous optic neuropathy—highlighting the relevance of this model for studies of glaucoma. Our preliminary functional genomic analysis suggested that GLIS1 interacts with GLIS3 and FOXC1, transcription factors previously implicated in elevated IOP, to regulate gene expression in TM cells. Moreover, reduced or increased GLIS1 activity can impair the integrity of ocular drainage tissues. Using unique mouse models, genetic and functional genomic approaches, and in vitro assays, we propose to characterize the GLIS1-dependent transcriptional regulatory network and determine its role in homeostasis and dysfunction of ocular drainage tissue. In Aim 1, we will test the hypothesis that increased GLIS1 expression contributes to POAG-associated ocular drainage tissue defects and determine whether the POAG-associated variants we identified in GLIS1 enhancer regions increased its transcriptional activity in primary human TM cells. We will also test whether GLIS1 overexpression in the mouse TM leads to high IOP and ocular drainage tissue defects similar to those in POAG. Finally, we will assess the potential role of dexamethasone and TGFβ2, previously implicated in IOP elevation, as upstream regulators of GLIS1. In Aim 2, we will test for potential genetic interactions between Glis1 and Foxc1 and/or Glis3 in ocular drainage tissue homeostasis. We will determine whether mice heterozygous for null alleles of both Glis1 and Foxc1 or Glis1 and Glis3 develop TM defects and altered IOP regulation. In parallel, we will characterize the transcriptional program and molecular pathways implicated in TM maintenance and function. These studies will provide important mechanistic insight into ocular drainage tissue homeostasis and dysfunction and could reveal targets for therapies to manage glaucoma.

抽象的青光眼是全球失明的主要原因，是一个重大的公共卫生问题。在美国，它影响到2050年，超过270万人及其患病率将增加到730万。防止青光眼或减慢其进展。主要的危险因素是高眼压（IOP），通常是由于水性幽默（AQH）出口受损。但是，所涉及的基因和途径知之甚少。我们已经确定了编码转录因子GLIS1的GLIS1，作为原发性开放的易感基因角度青光眼（POAG），表明Glis1 - / - 小鼠具有青光眼的病理生理标志。我们还发现GLIS1主要在小梁网（TM）中表达，这是该的关键组成部分调节AQH出口的眼部排水组织，Glis1 - / - 小鼠暴露了进行性TM变性，导致高IOP和青光眼的视神经神经病 - 高显示该模型与研究的相关性青光眼。我们的初步功能基因组分析表明，Glis1与Glis3相互作用 FOXC1，先前在IOP升高中实现的转录因子，以调节TM细胞中的基因表达。此外，降低或增加的GLIS1活性会损害眼部排水组织的完整性。使用唯一小鼠模型，遗传和功能基因组方法以及体外测定，我们建议表征依赖GLIS1的转录调节网络，并确定其在稳态和功能障碍中的作用眼部排水组织。在AIM 1中，我们将检验以下假设，即增加GLIS1表达有助于与POAG相关的眼引流组织缺陷，并确定与POAG相关的变体我们是否在GLIS1增强子区域中鉴定出来，增加了原代人TM细胞中其转录活性。我们将还测试小鼠TM中的GLIS1过表达是否导致高IOP和眼部排水组织缺陷与Poag中的人相似。最后，我们将评估地塞米松和TGFβ2的潜在作用，以前在IOP高程中实施，作为GLIS1的上游调节器。在AIM 2中，我们将测试潜在的通用眼部排水组织稳态中GLIS1和FOXC1和/或GLIS3之间的相互作用。我们将确定 Glis1和Foxc1和Glis1和Glis3的无效等位基因的杂合子是否会产生TM缺陷，并且 IOP调节改变了。同时，我们将表征转录程序和分子途径在TM维护和功能中实现。这些研究将为眼睛提供重要的机理洞察力排水组织稳态和功能障碍，可以揭示用于治疗青光眼的疗法的靶标。