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: 10450171
• 负责人: MARGARET M DEANGELIS
• 金额: $ 57.42万
• 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/10450171
• 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; 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 Diseases; Retinal Pigments; Role; Sampling; Signal Transduction; Structure; Structure of retinal pigment epithelium; Surface; 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; 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协调。所有 视网膜对正常的图像感知是最重要的。这是一个 衬在眼睛后部表面上的精确分层结构，包括 数以百万计的细胞聚集在一起形成一个紧密的网络。视神经 连接视网膜和大脑视网膜不仅接收光线， 处理它，并将下游信号传递到中脑和丘脑。 当视网膜发生病变，如老年性黄斑变性， (AMD)不幸的结果可能是失明，这是最可怕的残疾。 在AMD的遗传学方面的进展已经是实质性的，但是这些基因的翻译是不可能的。 结果到达诊所的速度很慢。延迟的原因包括缺乏 合适的动物模型进行功能遗传学，因为解剖 与人类的差异，对特定细胞类型的了解不足 参与AMD的启动和对人类视网膜病变的不完全理解， 生物学评估AMD的早期病理是否影响不同的细胞， 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）已经使得测量基因 在单个细胞中表达，为探索细胞异质性铺平了道路。 与亚拉巴马眼库合作，我们将深入采集人类视网膜样本， 细胞和RPE/脉络膜，充分表征细胞的多样性，并阐明功能 AMD全基因组关联研究结果的作用。我们建议 的目标。Aim 1将从20只眼睛中生成单个和批量RNA-seq数据 健康成人，24例早期/中期AMD和6例GA供体。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 将这些结果与AMD GWAS整合，以确定靶点 基因.这些开创性的研究利用新颖的方法和跨学科的 专门知识来表征人类视网膜中细胞类型特异性基因表达， 支持组织。通过对四种细胞图谱的详细表征， 在人类眼睛的地理区域，我们的研究将提供新的见解细胞， 类型特异性功能，可以为AMD的精确治疗靶向提供动力。