Molecular Mechanisms of Exfoliation Glaucoma

剥脱性青光眼的分子机制

基本信息

批准号：
10672918
负责人：
MICHAEL A HAUSER
金额：
$ 57.42万
依托单位：
DUKE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-09-01 至 2024-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10672918
关键词：
Accounting Address Affect Anterior Anterior eyeball segment structure Base Pairing Behavior Behavior Control Binding Biological Biological Assay Biological Models CD44 gene Cell Culture Techniques Cells Cellular biology Clinical Complex Confusion DNA Data Data Set Deposition Development Disease Event Exposure to Extracellular Matrix Eye diseases Feedback Functional disorder Gene Expression Genes Genetic Glaucoma Human Knowledge Mass Spectrum Analysis Measures Mechanics Mediating Messenger RNA Modification Molecular Molecular and Cellular Biology Monitor Morphology Names Ocular Hypertension Open-Angle Glaucoma Organ Culture Techniques Outcomes Research Pathway interactions Perfusion Pharmacology Physiologic Intraocular Pressure Physiology Primary Open Angle Glaucoma Promoter Regions Protein-Lysine 6-Oxidase Proteins RNA Splicing Regulation Reporting Research Resistance Risk Role Signal Pathway Signal Transduction Single Nucleotide Polymorphism Stretching Structure Structure of sinus venosus of sclera Testing Time Tissues Trabecular meshwork structure Untranslated RNA Variant Vision anterior chamber cell behavior cell type design disorder risk experimental study eye chamber genetic variant genome wide association study human tissue knock-down lens mechanotransduction monolayer novel therapeutics overexpression physical property promoter protective effect protein expression therapeutic target transcriptome

项目摘要

.ABSTRACT – Hauser/Stamer R01 . Pseudoexfoliation (PEX) glaucoma is a blinding form of glaucoma clinically characterized by insoluble protein deposits in the anterior chamber of the eye, and it is the most common identifiable secondary form of OAG, accounting for ~7 million cases worldwide. Genome wide association studies have identified genetic variants in the lysyl oxidase-like-1 (LOXL1) locus that are strongly associated with risk of PEX. Unfortunately, the functional mechanisms by which this locus contribute to PEX glaucoma are unknown. Our recent report characterized a long non-coding RNA (denoted herein as PEXpress) within the LOXL1 locus. We found that genetic variants alter the promoter strength of PEXpress, and are strongly associated with PEX glaucoma. We have extended these observations in preliminary experiments, revealing that knock down of PEXpress changes the expression of hundreds of downstream target genes, and analysis reveals modifications in pathways that lead to ocular hypertension including extracellular matrix remodeling and mechanotransduction. Using unbiased mass spectrometry and direct binding assays, we discovered that PEXpress specifically binds to the mRNA processing protein, hnRNPL via a 14 base pair binding region in PEXpress, and that endogenous hnRNPL complexes with endogenous PEXpress in cell culture. Based upon these observations, we hypothesize that alterations in the PEXpress/hnRNPL complex result in the dysregulation of downstream target genes, leading to altered trabecular meshwork (TM) and Schlemm’s canal (SC) cell biology, conventional outflow function and ultimately, increased intraocular pressure. To test this hypothesis, we have constructed three specific aims. In the first aim, we will determine the role of the PEXpress/hnRNPL complex on gene and protein regulation in human conventional outflow cells. Aim 2 is designed to determine the functional effects of the PEXpress/hnRNPL complex on human conventional outflow cells. Aim 3 will determine the effects of PEXpress on outflow facility using perfused human anterior segments in organ culture. As outcomes of this research we expect to (i) identify gene and protein targets plus signaling pathways regulated by PEXpress /hnRNPL complex in cell types responsible for the regulation of outflow resistance, (ii) identify role of PEXpress/hnRNPL in TM and SC cell signaling, contractility and barrier function, plus (iii) determine the role of PEXpress on conventional outflow function in an intact model system.

.摘要 – 豪瑟/斯塔默 R01 。假性剥脱性 (PEX) 青光眼是一种致盲性青光眼，临床特征为不溶性蛋白质沉积在眼前房，它是最常见的可识别的继发性 OAG 形式，全球范围内约 700 万例病例的全基因组关联研究已确定了基因变异。不幸的是，赖氨酰氧化酶样 1 (LOXL1) 位点与 PEX 风险密切相关。我们最近的报告描述了该基因座导致 PEX 青光眼的机制尚不清楚。我们发现 LOXL1 基因座内的长非编码 RNA（本文表示为 PEXpress）。改变 PEXpress 的启动子强度，并且与 PEX 青光眼密切相关。初步实验中的这些观察结果表明，PEXpress 的敲低会改变表达数百个下游靶基因，分析揭示了导致眼部疾病的途径的改变高血压，包括使用无偏质量进行细胞外基质重塑和机械转导。通过光谱测定和直接结合测定，我们发现 PEXpress 特异性结合 mRNA 加工蛋白质，hnRNPL 通过 PEXpress 中的 14 个碱基对结合区域，并且内源 hnRNPL 与基于这些观察，我们追踪了细胞培养中的内源性 PEXpress 的变化。 PEXpress/hnRNPL 复合物导致下游靶基因失调，导致小梁改变网状（TM）和施累姆氏管（SC）细胞生物学，常规流出功能并最终增加为了检验这一假设，我们制定了三个具体目标。确定 PEXpress/hnRNPL 复合物对人类常规基因和蛋白质调控的作用目的 2 旨在确定 PEXpress/hnRNPL 复合物对人类的功能影响。目标 3 将确定 PEXpress 对使用灌注人体的流出设施的影响。作为本研究的结果，我们期望 (i) 识别基因和蛋白质。靶标以及由 PEXpress /hnRNPL 复合物调节的细胞类型中的信号通路负责流出阻力的调节，(ii) 确定 PEXpress/hnRNPL 在 TM 和 SC 细胞信号传导、收缩性中的作用和屏障函数，加上 (iii) 确定 PEXpress 对完整模型中传统流出函数的作用系统。