Deconstructing and Modeling the Single Cell Architecture of the Age-Related Macular Degeneration Retina and RPE/Choroid

年龄相关性黄斑变性视网膜和 RPE/脉络膜的单细胞结构的解构和建模

• 批准号: 10674702
• 负责人: MARGARET M DEANGELIS
• 金额: $ 60.76万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-01 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10674702
• 关键词: Address; Adult; Affect; Age related macular degeneration; Alabama; Alleles; Alzheimer' s Disease; American; Anatomy; Animal Model; Animals; Architecture; Area; Atlases; Automobile Driving; Autopsy; Back; Biology; Blindness; Brain; Cancer Patient; Cell model; Cell physiology; Cells; Choroid; Clinic; Collaborations; Complex; Computer software; Data; Defect; Development; Diabetic Retinopathy; Disease; Disease Pathway; Eye; Eye Banks; Fright; Functional disorder; Future; Gene Expression; Genes; Genetic; Genomics; Geographic Locations; Health; Heterogeneity; Hour; Human; Image; In Vitro; Individual; Industry; Light; Location; Measurable; Measures; Metabolic; Methods; Microglia; Midbrain structure; Modeling; Molecular; Optic Nerve; Outcome; Pathologic; Pathology; Perception; Peripheral; Pharmaceutical Preparations; Phenotype; Population; Precision therapeutics; Process; Quantitative Trait Loci; Recovery; Retina; Retinal Degeneration; Retinal Diseases; Role; Sampling; Signal Transduction; Structure; Structure of retinal pigment epithelium; Surface; Technology; Testing; Thalamic structure; Tissues; Translations; Vision; Visualization; cell type; computerized tools; design; differential expression; disability; disease diagnostic; genome wide association study; improved; insight; macula; novel; open source; single-cell RNA sequencing; therapeutic target; transcriptome sequencing; transmission process; user-friendly; web site

Vision requires an orchestrated coordination between all parts of the eye. Of all the parts, the retina is the most vital for normal perception of an image. It is a precisely layered structure lining the surface of the back of the eye, comprising many millions of cells packed together in a tightly knit network. The optic nerve connects the retina with the brain. The retina not only receives light, but also processes it, and transmits downstream signals to the midbrain and the thalamus. When the retina becomes diseased as in age-related macular degeneration (AMD), the unfortunate result can be blindness which is the most feared disability. Progress in the genetics of AMD has been substantial, yet the translation of these results has been slow to reach the clinic. Reasons for this delay include lack of suitable animal models to perform functional genetics because of anatomical differences with humans, insufficient understanding about the specific cell types involved in the initiation of AMD and an incomplete understanding of human retinal biology. It is challenging to assess if the early pathology in AMD affects diverse cell populations versus highly specific cell types. Recent technologic breakthroughs in single-cell RNA-seq (scRNA-seq) have made it possible to measure gene expression in single cells, paving the way for exploring cellular heterogeneity. Collaborating with the Alabama Eye Bank, we will deeply sample human retinal cells and RPE/choroid, fully characterize cell diversity, and elucidate the functional roles of findings from genome- wide association studies for AMD. We propose the following aims. Aim 1 will generate single and bulk RNA-seq data from eyes of 20 healthy adults, 24 early/intermediate AMD and 6 GA donors. Aim 2 will characterize cell diversity and cell gene expression in normal human retina and RPE/choroid, and compare these results to AMD eyes. Aim 3 will infer cell-type specific eQTLs and integrate these results with AMD GWAS to identify target genes. These pioneering studies leverage novel methods and interdisciplinary expertise to characterize cell type-specific gene expression in human retina and supporting tissues. By detailed characterization of the cell atlases in four geographical areas in human eye, our study will provide novel insights into cell- type specific functions that can power precision therapeutic targeting of AMD.

视觉需要眼睛各部分之间的精心协调。其中 在这些部分中，视网膜对于正常感知图像来说是最重要的。它是一个 眼睛后部表面的精确分层结构，包括 数以百万计的细胞聚集在一个紧密相连的网络中。视神经 连接视网膜和大脑。视网膜不仅接收光线，还 处理它，并将下游信号传输到中脑和丘脑。 当视网膜患病（如年龄相关性黄斑变性）时 (AMD)，不幸的结果可能是失明，这是最可怕的残疾。 AMD 遗传学方面取得了巨大进展，但这些成果的转化 结果到达诊所的速度很慢。造成这种延迟的原因包括缺乏 由于解剖学原因，适合进行功能遗传学的动物模型 与人类的差异，对特定细胞类型的了解不足 参与 AMD 的发生以及对人类视网膜的不完全了解 生物学。评估 AMD 的早期病理是否影响不同的细胞具有挑战性 群体与高度特异性的细胞类型。最近的技术突破 单细胞 RNA-seq (scRNA-seq) 使测量基因成为可能 在单细胞中表达，为探索细胞异质性铺平了道路。 与阿拉巴马州眼库合作，我们将深入采集人类视网膜样本 细胞和 RPE/脉络膜，充分表征细胞多样性，并阐明功能 AMD 全基因组关联研究结果的作用。我们建议 以下目标。目标 1 将生成来自 20 个人眼睛的单个和批量 RNA-seq 数据 健康成年人、24 名早期/中期 AMD 和 6 名 GA 捐献者。目标2将 表征正常人视网膜中的细胞多样性和细胞基因表达 RPE/脉络膜，并将这些结果与 AMD 眼睛进行比较。目标 3 将推断细胞类型 特定的 eQTL 并将这些结果与 AMD GWAS 相结合以确定目标 基因。这些开创性的研究利用了新颖的方法和跨学科 表征人类视网膜中细胞类型特异性基因表达的专业知识 支持组织。通过四种细胞图谱的详细表征 人眼的地理区域，我们的研究将为细胞提供新的见解 类型特定功能可以为 AMD 的精准治疗提供动力。