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

项目3——细胞外和细胞内钙化的机制

基本信息

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
项目类别：
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&apos s Disease Apical Authorization documentation Autopsy Back Basal Cell Basic Science Binding Proteins Biological Models Blindness Blood Circulation Bruch&apos 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

项目摘要

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 摘要我们实验室的最新研究结果表明，细胞外和细胞内钙化的机制。细胞外钙化是年龄相关性黄斑变性（AMD）标志性病变的一个组成部分。富含磷酸钙矿物羟基磷灰石 (HAP) 的钙化沉积物积聚在下面的空间中眼睛后部的视网膜色素上皮 (RPE)，在这里吸收营养并清除不需要的物质物质通常在血液循环和 RPE 细胞之间交换。该过程可能导致快速进展为以不可逆转的视力丧失为特征的终末期疾病。与 AMD 相比，亚 RPE 沉积物的形成也与阿尔茨海默病等其他疾病有关 (AD) 根据最近的发现，我们提出了一种新的亚 RPE 沉积起始和增长模型，其中我们从临床和基础科学研究中得知，钙化起着关键作用。与蛋白质和脂质的相关相互作用也对疾病进展产生负面影响。我们了解到，了解不同的矿化形式如何导致疾病至关重要，但我们确实如此。尚不知道钙化是如何发生的以及细胞内和细胞外分子和途径有何贡献该项目将研究亚 RPE 空间中的矿物质沉积。使用我们开发的细胞模型系统来研究细胞外错位（异位）钙化我们还将开发一个显着改善的、富含矿物质的亚 RPE 矿床的起始和进展。新的 3D 微流体细胞模型用于研究钙化、亚 RPE 沉积物的形成、发展和 AMD 和 AD 的进展与脉络膜内皮及其变性有关。使用 AMD 和 AD 患者死后眼组织进行验证。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 之外，还有这些。治疗策略也适用于亚 RPE 沉积物发挥作用的其他疾病。