Project 3 - Mechanisms of extra- and intra-cellular calcification

项目3概要 细胞外和细胞内钙化的机制。我们实验室最近的研究结果表明， 细胞外钙化是年龄相关性黄斑变性（AMD）的标志性病变的组成部分。 富含磷酸钙矿物羟基磷灰石（HAP）的钙化沉积物积聚在 视网膜色素上皮细胞（RPE）在眼睛的后面，营养吸收和清除不需要的 这些物质通常在血液循环和RPE细胞之间交换。禁止这种交流 这一过程可能导致快速进展为以不可逆视力丧失为特征的终末期疾病。除了 从AMD开始，亚RPE存款形成也与其它疾病如阿尔茨海默病有关 （AD）。基于最近的研究结果，我们提出了一个新的模型，亚RPE存款启动和增长，其中 钙化起着关键作用。我们从临床和基础科学研究中知道，钙化， 与蛋白质和脂质相关的相互作用对疾病进展具有负面影响。我们也 了解到了解不同的矿化形式如何导致疾病至关重要。但我们 还不知道钙化是如何发生的，以及细胞内和细胞外的分子和途径对钙化的影响。 RPE下空间的矿物质沉积。该项目将研究细胞和系统的贡献者， 细胞外错位（异位）钙化使用细胞模型系统，我们开发的研究， 含矿物质的亚RPE沉积物的形成和发展。我们还将开发一个显著改进的， 一种新的3D微流体细胞模型，用于研究钙化，作为亚RPE存款的形成，发展和 AMD和AD的进展与脉络膜内皮及其变性有关。结果将 使用来自AMD和AD患者的人死后眼组织进行验证。验证和 使用2D和3D细胞培养系统的实验计划将为开发新的 干预策略以RPE下沉积物的钙化为中心。除了AMD和AD之外， 治疗策略也适用于其中亚RPE沉积物起作用的其它疾病。

PROJECT 3 SUMMARY Mechanisms of extra- and intra-cellular calcification. Recent findings from our laboratory showed that extracellular calcification is an integral part of the hallmark lesions of age-related macular degeneration (AMD). Calcified deposits rich in the calcium phosphate mineral hydroxyapatite (HAP) accumulate in the space under the retinal pigment epithelium (RPE) in the back of the eye, where nutrient uptake and clearance of unwanted material usually are exchanged between the blood circulation and the RPE cells. Inhibition of this exchange process can lead to rapid progression to an end-stage disease characterized by irreversible visual loss. Apart from AMD, sub-RPE deposit formation is also associated with other diseases such as Alzheimer’s disease (AD). Based on recent findings, we proposed a new model for sub-RPE deposit initiation and growth where calcification plays a pivotal role. We know from clinical and basic science studies that calcification, and the associated interactions with proteins and lipids, have a negative effect on disease progression. We also learned that understanding how different mineralization forms contribute to diseases is critical. However, we do not yet know how calcification occurs and what intra- and extracellular molecules and pathways contribute to mineral deposition in the sub-RPE space. This project will study the cellular and systemic contributors to extracellular mislocalized (ectopic) calcification using a cellular model system we developed to study the initiation and progression of the mineral-laden sub-RPE deposits. We will also develop a significantly improved, new 3D microfluidic cell model to study calcification, as sub-RPE deposit formation, development and progression of AMD and AD are linked to the choroidal endothelium and its degeneration. The results will be validated using human postmortem eye tissues from patients with AMD and AD. The validation and the experimental plan using the 2D and 3D cell culture systems will provide a robust foundation for developing new intervention strategies centered on the calcification of the sub-RPE deposits. Apart from AMD and AD, these treatment strategies will also be applicable for other diseases in which sub-RPE deposits play a role.