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

项目3——细胞外和细胞内钙化的机制

• 批准号: 10628930
• 负责人: Imre Lengyel
• 金额: $ 34.01万
• 依托单位: MEDICAL COLLEGE OF WISCONSIN
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-07-01 至 2028-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10628930
• 关键词: 3-Dimensional; Affect; Age related macular degeneration; Aging; Alzheimer' s Disease; Apical; Authorization documentation; Autopsy; Back; Basal Cell; Basic Science; Binding Proteins; Biological Models; Blindness; Blood Circulation; Bruch' s basal membrane structure; Cadaver; Calcium; Cell Culture System; Cell Culture Techniques; Cell model; Cells; Choroid; Clinical Sciences; Collection; Complex; Confocal Microscopy; Culture Media; Data; Dependovirus; Deposition; Development; Devices; Diphosphates; Disease; Disease Progression; Electron Microscopy; Endothelium; Ethics; Eye; Fluorescence; Foundations; Genes; Growth; Human; Hydroxyapatites; In Vitro; Intervention; Laboratories; Lead; Learning; Lesion; Link; Lipids; London; Maps; Methods; Microfluidic Microchips; Microfluidics; Minerals; Modeling; Molecular; Monitor; Nutrient; Ophthalmology; Pathway interactions; Patients; Play; Positioning Attribute; Primary Cell Cultures; Process; Proteins; Role; Sampling; Satellite Viruses; Serum; Structure of retinal pigment epithelium; Techniques; Tissues; Transfection; Universities; Validation; Visualization; calcification; calcification inhibitor; calcium phosphate; college; end stage disease; experimental study; extracellular; genetic makeup; human subject; human tissue; improved; in vivo; inorganic phosphate; laser capture microdissection; macromolecule; materials science; microvesicles; mineralization; multiple omics; overexpression; repository; single cell sequencing; three dimensional cell culture; three-dimensional modeling; treatment strategy; uptake; vector

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.

项目3摘要 细胞外计算的机制。我们实验室的最新发现表明 细胞外钙化是与年龄相关的黄斑变性（AMD）的标志性病变的组成部分。 计算出富含磷酸钙矿物羟基磷灰石（HAP）的沉积物积聚在空间中 眼睛背面的视网膜色素上皮（RPE），那里的营养吸收和清除不需要 通常在血液循环和RPE细胞之间交换材料。抑制这种交流 过程可能导致迅速发展为以视觉丧失不可逆转的特征的末期疾病。除外 从AMD中，子RPE沉积形成也与其他疾病（例如阿尔茨海默氏病）有关 （广告）。根据最近的发现，我们提出了一个针对子RPE存款计划和增长的新模型 钙化起关键作用。我们从临床和基础科学研究中知道，钙化以及 与蛋白质和脂质相关的相互作用对疾病进展有负面影响。我们也是 了解到，了解不同的矿化形式如何对疾病产生贡献至关重要。但是，我们这样做 尚不知道计算是如何发生的，哪些细胞内分子和途径有助于 子RPE空间中的矿物沉积。该项目将研究细胞和全身性贡献者 我们开发的用于研究的细胞外（异位）计算（异位）计算 矿物质亚rpe沉积物的起始和进展。我们还将发展出明显的改进， 新的3D微流体细胞模型研究计算，作为子RPE沉积的形成，发育和 AMD和AD的进展与脉络膜内皮及其变性有关。结果将是 使用AMD和AD患者的人类后眼组织进行验证。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基础提供强大的基础 干预策略以子RPE沉积物的计算为中心。除了AMD和AD，这些 治疗策略也将适用于其他疾病，其中亚RPE沉积物起着作用。