Single-Cell Transcriptomic Analysis of Human Retina

人类视网膜的单细胞转录组分析

基本信息

批准号：
10159930
负责人：
Mingyao Li
金额：
$ 53.49万
依托单位：
UNIVERSITY OF PENNSYLVANIA
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-05-01 至 2023-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10159930
关键词：
Address Adult Affect Age related macular degeneration Alabama Alleles Alzheimer&apos s Disease Anatomy Animal Model Atlases Automobile Driving Autopsy Back Biology Blindness Brain Cell physiology Cells Cellular Morphology Choroid Communities Complex Computer software Data Diabetic Retinopathy Diagnostic Disease Eye Eye Banks Eye diseases Fright Future Gene Expression Gene Expression Profile Genes Genetic Genomics Geographic Locations Health Heterogeneity Hour Human Human Genetics Image Immunohistochemistry Laboratories Light Measures Methods Microglia Midbrain structure Mus Mutation Optic Nerve Pathologic Pattern Perception Phenotype Population Precision therapeutics Process Recovery Retina Retinal Diseases Role Sampling Signal Transduction Stains Structure Structure of retinal pigment epithelium Surface Testing Thalamic structure Thick Thinness Tissues Translations Vision Visualization cell type computerized tools deep learning denoising disability genome wide association study improved insight learning strategy macula medical specialties millimeter novel open source precision medicine protein expression single-cell RNA sequencing success therapeutic target transcriptome sequencing transcriptomics user-friendly web app web site

项目摘要

PROJECT SUMMARY Vision, the most important of the human senses, occupies 25% of the brain function. It 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, the unfortunate result is blindness, which is the most feared disability. Diseases that affect the retina are complex because of the diverse number of cell types and total number of cells involved. It remains challenging to assess if pathological phenotypes affect diverse cell populations versus highly specific cell types. While advances in retinal disease diagnostics have progressed rapidly, treatments for retinal diseases directed at primary genetic defects have progressed slowly. Despite major successes in genetics, the vision community is lagging behind the advances in precision medicine occurring in other specialties. Modest progress is due in part to an incomplete understanding of human retinal biology. Anatomical differences between humans and commonly used animal models have severely hindered the translation of results from laboratory to human health. Therefore, there is an urgent need to collect and analyze retinal cells from human eyes to advance our understanding of human retinal diseases and assess the cell type conservation between mouse and human. 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, fully characterize cell diversity, and elucidate the functional roles of findings from genome- wide association studies for retinal diseases. We propose the following aims. Aim 1 will generate scRNA-seq data from eyes of 20 healthy adult human donors, and produce de-noised gene expression data for downstream analyses. Aim 2 will characterize cell diversity in human retina and supporting tissues, and validate novel cell type-specific marker genes by immunohistochemistry. Aim 3 will infer cell type compositions and allele-specific gene expression in each cell type by integrating scRNA-seq and bulk RNA-seq data from normal human eyes. 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 retinal diseases.

项目摘要视觉是人类最重要的人，占据了大脑功能的25％。它需要一个眼睛各部位之间的协调协调。在所有零件中，视网膜对于正常而言是最重要的对图像的感知。这是一个精确的分层结构，衬有眼睛背面的表面，包括数百万个单元在紧密编织的网络中包装在一起。视神经将视网膜与脑。视网膜不仅接收光，还可以对其进行处理，并将下游信号传输到中脑和丘脑。当视网膜患病时，不幸的结果是失明，就是最担心的残疾。影响视网膜的疾病由于细胞数量的不同而变得复杂涉及的细胞类型和总数。评估病理表型是否影响各种细胞种群与高度特异性的细胞类型。虽然视网膜疾病诊断的进展具有进展迅速，针对原发性遗传缺陷的视网膜疾病的治疗方法已缓慢发展。尽管遗传学取得了重大成功，但愿景社区仍在精确的进步落后其他专业中发生的药物。适度的进步部分是由于对人类视网膜生物学。人类和常用动物模型之间的解剖学差异具有严重阻碍了实验室结果向人类健康的转化。因此，迫切需要收集和分析视网膜细胞从人眼中提高我们对人类视网膜疾病的理解并评估小鼠和人之间的细胞类型保护。最近的技术突破单细胞RNA-SEQ（SCRNA-SEQ）使得在单细胞中测量基因表达，铺路探索细胞异质性的方法。与阿拉巴马州眼库合作，我们将深入采样人类视网膜细胞完全表征细胞多样性，并阐明了基因组发现的功能作用视网膜疾病的广泛关联研究。我们提出以下目标。 AIM 1将产生scrna-seq 来自20个健康的成年人类捐助者的眼睛的数据，并产生含糊不清的基因表达数据下游分析。 AIM 2将表征人视网膜和支撑组织中的细胞多样性，以及通过免疫组织化学验证新型细胞类型特异性标记基因。 AIM 3将推断细胞类型组成通过整合SCRNA-SEQ和BOLK RNA-SEQ数据中的每种细胞类型中的等位基因特异性基因表达正常的人眼睛。这些开创性的研究利用新颖的方法和跨学科专业知识表征人视网膜中细胞类型特异性基因表达和支持组织。通过详细在人眼中四个地理区域中细胞图谱的表征，我们的研究将提供新颖对细胞类型特定功能的见解，可以为视网膜疾病的精确治疗靶向启动。