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.

项目总结/摘要 剥脱综合征是一种全身性弹性纤维疾病。这种疾病在眼睛中的表现是 假性剥脱性青光眼（XFG）是青光眼最常见的可识别原因， 这种疾病的特征是淀粉样沉积物的形成， 由人眼前段的各种组织造成。特别是，材料更加突出 沉积在透镜的前表面上、虹膜和透镜之间、瞳孔边缘处。材料 很容易被眼科医生在定期检查中识别出来，并被称为“头皮屑样 材料”。它的存在是疾病诊断的基础。人们普遍认为， 在瞳孔的打开和关闭期间虹膜和透镜之间施加的摩擦导致移位 并将材料释放到房水中。在该流体流出期间，材料被携带到 小梁网，导致堵塞和随之而来的IOP升高。 对材料组成的蛋白质组学研究揭示了许多组分的存在。其中， 已知某些蛋白质，如β-淀粉样蛋白（CLU）和ApoE，与β-淀粉样蛋白斑块有关 阿尔茨海默病的特征。迄今为止，对XFG的研究还不多， 与XFG有关，LOXL 1是第一个也是最好的研究。但我们对这是如何发生的知之甚少 材料形成，并且没有再现其形成的尝试。此外，XFM还没有 在任何动物物种中观察到，包括非人类灵长类动物。 在本申请中，我们提出解决这一需求。因为这种物质只在人类体内形成，我们设计了 一种对人类透镜进行研究的新策略，更准确地说，是在最前部区域， 临床观察材料。透镜的这一区域通常由眼科医生剥离（并丢弃 白内障手术中的外科医生称为“透镜囊”（LC），它包括14 µm透镜囊 下面是单层上皮细胞。因此，我们将开发和表征这些器官型 从废弃的外科LC培养物中开始研究XFM如何形成。我们将重点关注两大 相关组件，CLU和LOXL 1。我们将通过基因转移来调节它们的生产，并将它们暴露出来， 不仅对XFG应激源，而且对透镜近端组织，特别是虹膜分泌的条件培养基 色素上皮（IPE）。我们将通过共聚焦显微镜和它们的作用来评估XFG样沉积物的形成。 通过免疫组织化学观察对不溶性ECM中弹性蛋白和胶原蛋白网络的影响。我们有首字母缩写 可行性数据，尽管需要克服重大挑战，但我们认为，新系统可以 为该领域提供了宝贵的工具，并提供了有关该疾病的重要信息。的理解 这项研究所获得的也将打开大门，寻找具体的治疗假性剥脱 青光眼