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.

项目总结 老年性黄斑变性(AMD)是世界范围内导致失明的主要原因之一。失调症 视网膜色素上皮(RPE)是AMD的主要早期事件之一。RPE细胞发挥了许多作用 在视觉中的重要作用，并有助于保持视网膜的健康和完整性，这一层的缺陷导致 光感受器细胞的进行性退化。AMD的特征包括一种名为玻璃体的蛋白质沉积 位于视网膜色素上皮(RPE)下面，可演变为地理性萎缩或脉络膜 导致光感受器丧失的新生血管。虽然有治疗方法可以延缓糖尿病的进程 某些形式的AMD，一种阻止不可避免的视力丧失的根治方法并不存在。中国的一项迫切需要 FIELD是研究AMD早期特征的较好模型，如玻璃体形成和对致病因素的识别 机制是必要的。考虑到AMD的多因素性质，这是一个难题。众所周知， 光感受器和视网膜色素上皮(RPE)细胞关键依赖于严格控制的脂质 维持功能的动态平衡。每天，大约十分之一的棒外节段(OS)被脱落和 被RPE吞噬，作为视觉循环的一部分，RPE循环膜。考虑到大量的 细胞膜，主要由每天循环使用的脂类和蛋白质组成，有可能是脂类缺陷 新陈代谢可能至少参与了疾病病理的某些部分。在这个项目中，我们建议研究 作为年龄相关性眼病研究2的一部分，患者来源的人视网膜色素上皮细胞的脂类组学 (AREDS2)，旨在确定AMD的易感因素、临床特征和预后指标。我们 建议从这项研究中区分IPSC品系，评估品系之间的潜在变异性，并评估 转录差异。然后，我们将评估“疾病”与对照品系的脂肪含量的差异。 寻找潜在的疾病生物标志物。最后，我们将通过以下方式探讨行之间的功能差异 进行吞噬试验。我们希望共同识别新的疾病相关生物标志物，这些生物标志物可能 最终导致治疗这种毁灭性疾病的新的治疗选择。