Molecular Mechanisms of Exfoliation Glaucoma

剥脱性青光眼的分子机制

• 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
• 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

.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.

.abstract - Hauser/Stamer R01。 假脱糖粉（PEX）青光眼是一种盲目的青光眼临床表征，其特征是不溶性蛋白 在眼前腔室中的沉积物，它是OAG的最常见识别次级形式， 全球约有700万个案件。基因组广泛的关联研究已经确定了遗传变异 与PEX风险密切相关的赖氨酸氧化物样-1（LOXL1）基因座。不幸的是，功能 该基因座对PEX青光眼促进的机制尚不清楚。我们最近的报告描述了 LOXL1基因座中的长非编码RNA（本文表示为pexpress）。我们发现遗传变异 改变pexpress的启动子强度，并与PEX青光眼密切相关。我们已经扩展了 初步实验中的这些观察结果表明，击倒pexpress改变了表达 数百个下游靶基因和分析揭示了导致眼的途径的修改 高血压包括细胞外基质重塑和机械转导。使用公正的质量 光谱法和直接结合测定法，我们发现pexpress特异性结合了mRNA加工 蛋白质，通过14个碱基对结合区域中的hnrnpl，以及内源性HNRNPL复合物与 细胞培养中的内源性pexpress。基于这些观察结果，我们假设 pexpress/hnrnpl复合物导致下游靶基因失调，导致小梁改变 网格工作（TM）和Schlemm的运河（SC）细胞生物学，常规出口功能，最终增加 眼压。为了检验这一假设，我们构建了三个具体目标。在第一个目标中，我们将 确定pexpress/hnrnpl复合物对人类常规的基因和蛋白质调节的作用 出口细胞。 AIM 2旨在确定PEXPRESS/HNRNPL复合物对人的功能效应 常规的出口细胞。 AIM 3将确定Pexpress使用灌注的人类对出口设施的影响 器官培养的前段。作为这项研究的结果，我们期望（i）识别基因和蛋白质 目标加上由PEXPRESS /HNRNPL复合物调节的信号通路，负责细胞类型 调节出口电阻，（ii）确定pexpress/hnrnpl在TM和SC细胞信号，收缩力中的作用 和障碍功能，加上（iii）确定pexpress在完整模型中传统输出函数的作用 系统。