Mechanisms of Formation of Pseudoexfoliation Material on Human Surgical Lens Capsules

人体手术晶状体囊上假性剥脱材料的形成机制

• 批准号: 9808397
• 负责人: Teresa Borras
• 金额: $ 23.33万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 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 2021-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9808397
• 关键词: Address; Affect; Alzheimer' s Disease; Amyloid; Amyloid beta-Protein; Amyloid deposition; Animal Diseases; Animals; Anterior; Anterior Surface of the Lens; Apolipoprotein E; Aqueous Humor; Awareness; Back; Binding Proteins; Blindness; Cataract; Cataract Extraction; Cells; Cessation of life; Characteristics; Ciliary Body; Clinical; Complementary DNA; Confocal Microscopy; Contracts; Country; Data; Deposition; Development; Diagnostic; Disease; Elastic Fiber; Elastin; Environmental Risk Factor; Epithelial Cells; Epithelium; Exfoliation Syndrome; Exposure to; Extracellular Matrix; Eye; FBN1; Fiber; Friction; Gene Delivery; Gene Proteins; Gene Transfer; Genes; Glaucoma; Grant; Histologic; Human; Immunohistochemistry; Iris; Laboratories; Lead; Left; Link; Liquid substance; Logistics; Mechanics; Modeling; Molecular Chaperones; Nature; Operative Surgical Procedures; Ophthalmic examination and evaluation; Ophthalmologist; Ophthalmology; Outcome; Oxidative Stress; Pathologic; Patients; Pharmaceutical Preparations; Physiologic Intraocular Pressure; Physiological; Pigment Epithelium; Play; Production; Proteins; Proteomics; Pupil; Reagent; Resistance; Retinal Ganglion Cells; Role; Senile Plaques; Slide; Specimen; Stress; Surface; Surgeon; System; Systemic disease; Testing; Time; Tissues; Trabecular meshwork structure; Tropoelastin; Viral Vector; Visual Fields; amyloid formation; anterior chamber; aqueous humor flow; base; capsule; experience; experimental study; fibrillin; high intraocular pressure; high reward; high risk; interest; lens; lens capsule; nonhuman primate; novel strategies; optic nerve disorder; pressure; protein complex; small hairpin RNA; stressor; sulfated glycoprotein 2; tissue culture; tool

PROJECT SUMMARY/ ABSTRACT Exfoliation syndrome is a systemic disease of the elastic fibers. The manifestation of this disease in the eye is Pseudoexfoliation Glaucoma (XFG), which is the most common identifiable cause of glaucoma, the most aggressive, and the one harder to treat. The disease is characterized by the formation of amyloid-like deposits by various tissues of the anterior segment of the human eye. In particular, the material is more prominently deposited on the anterior surface of the lens, between the iris and the lens, at the pupillary border. The material is easily recognized by the ophthalmologist during a regular examination and it is dubbed as “a dandruff-like material”. Its presence is the base for the diagnostic of the disease. It is widely accepted that the mechanical friction exerted between the iris and the lens during the opening and closing of the pupil, leads to dislodgement and release of the material into the aqueous humor. During the outflow of this fluid, the material is carried to the trabecular meshwork, causing clogging and a consequent elevated IOP. Proteomics on the composition of the material revealed the presence of a number of components. Among them, some, such as Clusterin (CLU) and ApoE, have been known to be associated with the β-amyloid plaques characteristic of Alzheimer disease. No many studies on XFG have been conducted To date, seven genes have been linked to the XFG, with LOXL1 being the first and best studied. But very little is known about how this material is formed and no attempts to reproduce its formation are available. Also, the XFM has not been observed in any animal species, including nonhuman primates. In this application, we propose to address this need. Because this material is only formed in humans, we devised a novel strategy to conduct the study on human lens, more precisely on the most anterior region, where the material is clinically observed. This region of the lens is routinely peeled off (and discarded) by ophthalmology surgeons during cataract surgery. Termed “lens capsule” (LC), it comprises the 14 µm lens capsule together with the single layer of epithelial cells underneath. Thus, we will develop and characterize these organotypic cultures form the discarded surgical LCs to begin studying how the XFM is formed. We will focus on two major relevant components, CLU and LOXL1. We will modulate their production by gene transfer, and expose them, not only to XFG stressors, but to conditioned media secreted by the lens proximal tissues, specially by the Iris Pigment Epithelium (IPE). We will evaluate formation of XFG-like deposits by confocal microscopy and their effect on the elastin and fibrillin networks in the insoluble ECM by immunohistochemistry. We have initial feasibility data and although important challenges need to be overcome, we believe that the new system could provide an invaluable tool to the field and render important information about the disease. The understanding gained by this study will also open the door to the search of specific treatments of the pseudoexfoliation glaucoma.

项目摘要/摘要 去角质综合征是弹性纤维的全身性疾病。这种疾病在眼中的表现是 假脱糖曲瘤青光眼（XFG），这是青光眼最常见的原因，最常见的原因是最常见的原因 积极进取，一个更难治疗的人。该疾病的特征是形成淀粉样蛋白的沉积物 通过人眼前部的各种组织。特别是，该材料更为突出 沉积在晶状体的前表面，虹膜和镜头之间，瞳孔边界。材料 在常规检查中很容易被眼科医生认识，并且被称为“类似头皮屑的样子 材料”。它的存在是疾病诊断的基础。 在学生开放和关闭期间，虹膜和镜头之间施加了摩擦，导致脱落 并将材料释放到水性幽默中。在这种流体的出口期间，将材料携带到 小梁网架，引起堵塞，随之而来的IOP升高。 材料组成的蛋白质组学揭示了许多成分的存在。他们之中， 已知有些（例如簇蛋白（CLU）和APOE）与β-淀粉样斑块有关 阿尔茨海默氏病的特征。迄今为止，还没有进行过关于XFG的研究，七个基因具有 我们与XFG链接，LOXL1是第一个也是最好的研究。但是对这是什么知之甚少 形成材料，没有尝试复制其形成的尝试。另外，XFM还没有 在任何动物物种中都观察到，包括非人类隐私。 在此应用程序中，我们建议满足这一需求。因为这种材料仅在人类中形成，所以我们设计了 对人晶状体进行研究的一种新型策略，更准确地说是在最前部区域 临床观察到材料。镜头的这个区域通常被眼科剥离（并丢弃） 白内障手术期间的外科医生。称为“镜头胶囊”（LC），共同包含14 µm镜头胶囊 下面的上皮细胞的单层。那就是我们将发展并描述这些有机物 培养物形成废弃的手术LC，开始研究XFM的形成方式。我们将专注于两个主要 相关组件，clu和loxl1。我们将通过基因转移来调节它们的生产，并暴露它们， 不仅要XFG压力源，而且要针对透镜代理组织分泌的条件培养基，特别是由虹膜 色素上皮（IPE）。我们将评估通过共聚焦显微镜及其的形成XFG样沉积物的形成 免疫组织化学对不溶性ECM中弹性蛋白和原纤维蛋白网络的影响。我们有初始 可行性数据，尽管需要克服重要的挑战，但我们认为新系统可以 为该领域提供宝贵的工具，并提供有关该疾病的重要信息。理解 这项研究获得的还将为寻找伪exfoliation的特定治疗方法打开大门 青光眼。