Mechanisms of Formation of Pseudoexfoliation Material on Human Surgical Lens Capsules

项目摘要/摘要 剥脱综合征是一种全身性弹力纤维疾病。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，最 攻击性强，也是最难治疗的。该病的特征是形成淀粉样沉积。 通过人眼前段的各种组织。特别是，材料更加突出 位于晶状体前表面，虹膜和晶状体之间，在瞳孔边缘。材料 在常规检查中很容易被眼科医生辨认出来，被称为“头皮屑样” 材料“。它的存在是诊断该病的基础。人们普遍认为，机械设备 在瞳孔打开和关闭过程中，虹膜和晶状体之间产生的摩擦会导致移位。 并将材料释放到房水中。在这种液体流出的过程中，物质被带到 小梁网状结构，造成阻塞，继而导致眼压升高。 蛋白质组学对材料的组成进行了分析，发现存在许多成分。其中， 已知一些与β-淀粉样斑块有关的物质，如聚集素和载脂蛋白E 阿尔茨海默病的特征。到目前为止，对XFG的研究还不多，有7个基因 已与XFG联系在一起，LOXL1是第一个也是研究最深入的。但人们对这一点知之甚少 材料已经形成，没有任何复制其形成的尝试。此外，XFM还没有 在任何动物物种中都能观察到，包括非人类灵长类动物。 在本申请中，我们建议解决这一需求。因为这种物质只在人类中形成，所以我们设计了 一种对人类晶状体进行研究的新策略，更准确地说，是在最前面的区域进行研究，在那里 材料是临床观察的。眼科医生通常会将晶状体的这一区域剥离(并丢弃)。 白内障手术中的外科医生。被称为“晶状体胶囊”(LC)，它由14微米的晶状体胶囊组成 下面是单层的上皮细胞。因此，我们将开发和表征这些器官类型 培养形成废弃的外科LC，开始研究XFM是如何形成的。我们将重点关注两个主要问题 相关组件、CLU和LOXL1。我们将通过基因转移来调节它们的生产，并将它们暴露出来， 不仅对XFG应激源，而且对晶状体近端组织，特别是虹膜组织分泌的条件介质 色素上皮(IPE)。我们将用共聚焦显微镜评价类XFG矿床的形成及其 免疫组织化学方法对不溶性ECM中弹性蛋白和纤维蛋白网络的影响。我们有首字母 虽然需要克服重大挑战，但我们认为，新系统可以 为实地提供宝贵的工具，并提供有关疾病的重要信息。理解 这项研究也将为寻找假性剥脱症的具体治疗方法打开大门 青光眼。

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.