Correlating Genomic AMD Risk Variants with Lipid Composition and Phagocytic Function of Patient-Derived Induced Pluripotent Stem Cell (iPSC)-derived Retinal Pigment Epithelium (RPE)

将基因组 AMD 风险变异与患者来源的诱导多能干细胞 (iPSC) 来源的视网膜色素上皮 (RPE) 的脂质成分和吞噬功能相关联

• 批准号: 10701841
• 负责人: Dorota Skowronska-Krawczyk
• 金额: $ 39.35万
• 依托单位: UNIVERSITY OF CALIFORNIA-IRVINE
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-30 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10701841
• 关键词: Age related macular degeneration; Antibodies; Apical; Back; Basal lamina; Biological Assay; Biological Markers; Blindness; Blood-Retinal Barrier; Cell Line; Cell Polarity; Cell membrane; Cells; Characteristics; Choroidal Neovascularization; Clinical; Clustered Regularly Interspaced Short Palindromic Repeats; Collecting Cell; Coupled; Culture Media; Data; Defect; Deposition; Development; Disease; Docosahexaenoic Acids; Drusen; Electroretinography; Electrospray Ionization; Endocytosis; Enzymes; Event; Exhibits; Exocytosis; Eye; Eye diseases; Fatty Acids; Fluorescence; Functional disorder; Gene Expression Profile; Genomics; Growth Factor; Health; Heterogeneity; Homeostasis; Human; Immunohistochemistry; In Vitro; Inflammatory; Inflammatory Response; Laboratories; Lead; Light; Link; Lipids; Liquid Chromatography; Measures; Membrane; Methods; Modeling; Molecular; Mutation; Nature; Pathogenesis; Pathology; Patients; Phagocytes; Phagocytosis; Photoreceptors; Pigments; Play; Polyunsaturated Fatty Acids; Predisposing Factor; Proteins; RPE65 protein; Recycling; Resistance; Retina; Rod Outer Segments; Role; Sampling; Signal Transduction; Source; Stains; Structure of retinal pigment epithelium; System; Tight Junctions; U-Series Cooperative Agreements; Vision; Western Blotting; age related; cell type; chromophore; complement C3 precursor; differential expression; experimental study; genetic variant; geographic atrophy; high risk; induced pluripotent stem cell; inflammatory modulation; knockout gene; lipid metabolism; lipidomics; mass spectrometer; molecular marker; molecular phenotype; novel; novel therapeutic intervention; novel therapeutics; participant enrollment; photoreceptor degeneration; potential biomarker; prognostic indicator; receptor binding; retinal rods; risk variant; tool; transcriptome sequencing; transcriptomics; visual cycle

PROJECT SUMMARY Age-related macular degeneration (AMD) is one of the leading causes of blindness worldwide. Dysfunction of the retinal pigment epithelium (RPE) is one of the primary early events during AMD. RPE cells play many important roles in vision and helps to maintain the health and integrity of the retina and defects in this layer lead to progressive degeneration of photoreceptor cells. Hallmarks of AMD include protein deposits called drusen which lie underneath the retinal pigment epithelium (RPE) and can evolve into geographic atrophy or choroidal neovascularization with consequent loss of photoreceptors. While treatments exist that slow down the course of some forms of AMD, a permanent cure to stop the inexorable loss of vision does not exist. A critical need in the field is a better model for early features of AMD such as drusen formation and recognition of disease-causing mechanisms is needed. This is a difficult problem given the multifactorial nature of AMD. It is well known that photoreceptor and retinal pigment epithelial (RPE) cells are crucially dependent on tightly controlled lipid homeostasis to maintain function. Each day, roughly a tenth of the rod outer segments (OS) are shed and phagocytized by the RPE, which recycle the membranes as part of the visual cycle. Given the vast amount of cell membrane, mostly comprised of lipids and proteins that is recycled daily, it is plausible that defects in lipid metabolism could be involved in at least some part of the disease pathology. In this project we propose to study lipidomics in human RPE cells from patient derived iPSCs as part of the Age-Related Eye Disease Study 2 (AREDS2) which aims to identify predisposing factors, clinical features, and prognostic indicators of AMD. We propose to differentiate iPSC lines from this study, assess potential variability between lines, and evaluate transcriptional differences. We will then assess differences in the lipid content of ‘disease’ versus control lines searching for potential biomarkers of disease. Lastly, we will explore functional differences between lines by performing phagocytosis assays. Together, we hope to identify novel disease-associated biomarkers that may ultimately lead to new therapeutic options for treating this devastating disease.

项目概要 年龄相关性黄斑变性（AMD）是全世界失明的主要原因之一。 视网膜色素上皮 (RPE) 是 AMD 中 RPE 细胞发挥多种作用的主要早期事件之一。 在视力中发挥重要作用，有助于维持视网膜的健康和完整性以及该层铅的缺陷 AMD 的特征包括称为玻璃膜疣的蛋白质沉积。 位于视网膜色素上皮 (RPE) 下方，可演变成地理性萎缩或脉络膜萎缩 新血管形成导致光感受器丧失，而现有的治疗方法可以减缓病程。 对于某些形式的 AMD，目前并不迫切需要永久治愈以阻止不可避免的视力丧失。 场是AMD早期特征的更好模型，例如玻璃疣的形成和致病因素的识别 鉴于 AMD 的多因素性质，这是一个难题。 光感受器和视网膜色素上皮 (RPE) 细胞很大程度上依赖于严格控制的脂质 每天，大约十分之一的视杆外节 (OS) 会脱落并脱落。 被 RPE 吞噬，作为视觉循环的一部分，RPE 会回收膜。 细胞膜主要由每天循环利用的脂质和蛋白质组成，因此脂质缺陷似乎是合理的 新陈代谢可能至少涉及疾病病理学的某些部分。在这个项目中，我们建议进行研究。 作为年龄相关眼病研究 2 的一部分，对来自患者 iPSC 的人类 RPE 细胞进行脂质组学研究 (AREDS2) 旨在确定 AMD 的诱发因素、临床特征和预后指标。 建议从本研究中区分 iPSC 细胞系，评估细胞系之间的潜在变异性，并评估 然后我们将评估“疾病”与对照系的脂质含量差异。 最后，我们将通过以下方式探索品系之间的功能差异。 我们希望共同进行吞噬作用测定，以确定可能的新型疾病相关生物标志物。 最终为治疗这种毁灭性疾病带来新的治疗选择。