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

基本信息

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
项目类别：
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&apos 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

项目摘要

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.

项目概要/摘要剥脱综合征是一种弹力纤维的全身性疾病，这种疾病在眼部的表现是。假性剥脱性青光眼 (XFG) 是青光眼最常见的可识别病因，也是最常见的青光眼病因。该疾病具有侵袭性，且较难治疗，其特点是形成淀粉样蛋白沉积物。尤其是人眼眼前节的各种组织材料更为突出。沉积在晶状体前表面、虹膜和晶状体之间、瞳孔边缘处。眼科医生在定期检查时很容易识别出这种现象，被称为“类似头皮屑的现象”。人们普遍认为，它的存在是诊断该疾病的基础。在瞳孔打开和关闭过程中虹膜和晶状体之间施加的摩擦会导致移位以及将物质释放到房水中。在该液体流出期间，物质被携带至眼房水。小梁网，导致堵塞并导致眼压升高。该材料组成的蛋白质组学揭示了其中许多成分的存在，其中一些，例如 Clusterin (CLU) 和 ApoE，已知与 β-淀粉样斑块有关迄今为止，尚未对 XFG 进行太多研究，有 7 个基因是阿尔茨海默病的特征。与 XFG 相关联，其中 LOXL1 是第一个也是研究最深入的，但人们对其如何进行了解却知之甚少。材料已形成，并且无法尝试重现其形成，而且 XFM 尚未可用。在任何动物物种中观察到，包括非人类灵长类动物。在此应用中，我们提出解决这一需求，因为这种材料仅在人类中形成，所以我们设计了这种材料。一种对人类晶状体进行研究的新策略，更准确地说是在最前部区域，其中眼科通常会通过临床观察将镜片的该区域剥离（并丢弃）。外科医生在白内障手术中将其称为“晶状体囊”(LC)，它由 14 µm 晶状体囊组成。下面是单层上皮细胞，因此，我们将开发并表征这些器官型。培养物形成废弃的手术 LC，开始研究 XFM 的形成方式，我们将重点关注两个主要方面。相关组件，CLU和LOXL1我们将通过基因转移调节它们的产生，并暴露它们，不仅针对 XFG 应激源，而且针对晶状体近端组织（特别是虹膜）分泌的条件介质我们将通过共焦显微镜及其沉积物评估 XFG 样沉积物的形成。通过免疫组织化学我们初步了解了不溶性 ECM 中弹性蛋白和原纤维蛋白网络的影响。可行性数据，尽管需要克服重要挑战，但我们相信新系统可以为该领域提供了宝贵的工具并提供有关该疾病的重要信息。这项研究获得的成果也将为寻找假性剥脱的具体治疗方法打开大门青光眼。