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

项目3摘要 细胞外和细胞内钙化的机制。我们实验室的最新发现表明 细胞外钙化是老年性黄斑变性(AMD)标志性病变的重要组成部分。 富含钙磷矿物羟基磷灰石(HAP)的钙化沉积物堆积在地下空间 位于眼球后部的视网膜色素上皮(RPE)，在那里营养物质的吸收和不需要的物质的清除 物质通常在血液循环和RPE细胞之间交换。对这种交流的抑制 这一过程可导致以不可逆转的视力丧失为特征的终末期疾病的快速进展。分开 在AMD中，亚RPE沉积也与其他疾病有关，如阿尔茨海默病 (Ad)。基于最近的发现，我们提出了一种新的亚RPE矿床形成和生长模型，其中 钙化起着关键作用。我们从临床和基础科学研究中了解到，钙化和 与蛋白质和脂质相关的相互作用，对疾病进展有负面影响。我们也 认识到了解不同矿化形式如何导致疾病是至关重要的。然而，我们有 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此有何贡献 RPE以下空间中的矿物沉积。该项目将研究细胞和系统因素对 使用我们开发的细胞模型系统研究细胞外错位(异位)钙化 富含矿物的次RPE矿床的形成和演化。我们还将开发一种显著改进的， 新的3D微流体细胞模型用于研究钙化作用，作为亚RPE矿床的形成、发展和 AMD和AD的进展与脉络膜内皮细胞及其变性有关。结果将是 使用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 干预策略以亚RPE沉积物的钙化为中心。除了AMD和AD，这些 治疗策略也将适用于RPE亚沉积起作用的其他疾病。

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.