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，在美国也是一个重大的公共卫生问题。超过 270 万人，到 2050 年患病率将上升至 730 万人。需要靶向治疗预防青光眼或减缓其进展的一个主要危险因素是高眼压（IOP），通常是由于高眼压（IOP）引起的。然而，人们对房水（AqH）流出受损的基因和途径知之甚少。我们已经鉴定出编码转录因子 GLIS1 的 GLIS1 作为原发性开放性遗传病的易感基因。角型青光眼 (POAG) 并表明 Glis1–/– 小鼠具有青光眼的病理生理学特征。还发现 Glis1 主要在小梁网 (TM) 中表达，小梁网是细胞的关键组成部分。眼部引流组织调节 AqH 流出，并且 Glis1–/– 小鼠表现出进行性 TM 变性，导致高眼压和青光眼性视神经病变——强调了该模型与研究的相关性我们的初步功能基因组分析表明 GLIS1 与 GLIS3 相互作用。 FOXC1 是先前与眼压升高有关的转录因子，可调节 TM 细胞中的基因表达。此外，GLIS1 活性的降低或增加会损害眼部引流组织的完整性。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 GLIS1 依赖性转录调控网络并确定其在稳态和功能障碍中的作用在目标 1 中，我们将检验 GLIS1 表达增加有助于的假设。 POAG 相关的眼部引流组织缺陷并确定 POAG 相关变异是否是我们所关注的 GLIS1 增强子中确定的区域增加了其在原代人类 TM 细胞中的转录活性。还测试小鼠 TM 中 GLIS1 过度表达是否导致高 IOP 和眼部引流组织缺陷与 POAG 中的类似，最后，我们将评估之前的地塞米松和 TGFβ2 的潜在作用。作为 GLIS1 的上游调节因子，与 IOP 升高有关。在目标 2 中，我们将测试遗传潜力。我们将确定 Glis1 和 Foxc1 和/或 Glis3 在眼引流组织稳态中的相互作用。 Glis1 和 Foxc1 或 Glis1 和 Glis3 无效等位基因杂合的小鼠是否会出现 TM 缺陷以及同时，我们将描述转录程序和分子途径的特征。这些研究将为眼部的机制提供重要的见解。引流组织稳态和功能障碍，并可能揭示青光眼治疗的目标。