Genetic Modifiers of Enhanced S-cone Syndrome –Role of the External Limiting Membrane

增强型 S 锥综合征的遗传修饰 — 外部限制膜的作用

基本信息

批准号：
10334439
负责人：
JUERGEN K. NAGGERT
金额：
$ 42.44万
依托单位：
JACKSON LABORATORY
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-02-01 至 2024-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10334439
关键词：
Address Adherens Junction Adult Affect Age related macular degeneration Alleles Apical Back Binding Blindness Cell membrane Cell physiology Cells Chemicals Cluster Analysis Cone Data Defect Development Diabetic Retinopathy Diffusion Disease Disease Pathway Disease Progression Disease model Dysplasia Enhanced-S cone syndrome Ethylnitrosourea Event Eye Eye diseases Failure Fundus Gene Mutation Gene Proteins Genes Genetic Genetic Screening Goals High-Throughput Nucleotide Sequencing Human Image Immunofluorescence Immunologic Immunoprecipitation Inherited Intercellular Junctions Knockout Mice Knowledge Lead Leber&apos s amaurosis Location Maintenance Membrane Modeling Molecular Muller&apos s cell Mus Mutagenesis Mutate Mutation Natural History Nature Night Blindness Optical Coherence Tomography Pathologic Pathology Pathway interactions Patients Phenotype Photoreceptors Population Process Proteins Public Health Research Retina Retinal Cone Retinal Degeneration Retinal Diseases Retinal Dysplasia Retinitis Pigmentosa Rod Role Stains Structure Testing Tight Junctions Variant Visual Acuity Visual impairment Yeasts bioinformatics tool cell type inherited retinal degeneration insight mouse model mutant novel photoreceptor degeneration precursor cell prevent recruit retinal progenitor cell single cell analysis single-cell RNA sequencing tool transcription factor transcriptome yeast two hybrid system

项目摘要

PROJECT SUMMARY/ABSTRACT Enhanced S-cone syndrome (ESCS) is an inherited retinal degeneration characterized by an increased number of S-cones, retinal dysplasia, and progressive photoreceptor degeneration leading to early night blindness and loss of visual acuity. The disease is caused by mutations in the rod-fate determining transcription factor NR2E3 in humans and the rd7 mouse model. The disease presentation is highly variable in human and dependent on genetic background in the mouse, demonstrating the existence of genetic modifiers. The mouse model demonstrates that the retinal dysplasia is intimately associated with the occurrence of breaks in the external limiting membrane (ELM), a network of adherens/tight junctions between Müller cell apical processes and photoreceptor inner segments. Fragmentation of the ELM is also observed in other retinal diseases associated with dysplasia, such as Leber's Congenital Amaurosis (LCA) due to mutations in crumbs1 (CRB1) and RP27, and in diabetic retinopathy. We have identified a genetic modifier that prevents the fragmentation of the ELM in both the rd7 mouse and the Nrl ko mouse (RP27) models. This discovery strongly suggests a causative role for the ELM fragmentation in the development of photoreceptor dysplasia and provides a tool with which to establish the mechanisms by which ELM breaks occur, how they lead to retinal dysplasia, and how this affects disease progression. In order to identify these mechanisms we will: 1) Determine the natural history of ELM junction formation and photoreceptor differentiation in wt and the rd7 model using marker analysis of cell junction proteins. 2) Identify the cell types involved in the ELM fragmentation by single cell RNAseq analysis. 3) Determine how the modifier protein alters recruitment of junctional proteins to the cell membrane. 4) Identify the molecular basis of additional genetic modifier strains that we have generated to gain further insight into the NR2E3 disease pathways.

项目摘要/摘要增强的S-Cone综合征（ESC）是一种遗传性视网膜变性，其特征是增加 S-cons，视网膜发育不良和进行性光感受器变性的数量导致清夜视力的失明和视力丧失。该疾病是由确定杆命令中的突变引起的人类和RD7小鼠模型中的转录因子NR2E3。疾病的表现在很大程度上变化人类和依赖小鼠的遗传背景，证明了遗传修饰剂的存在。小鼠模型表明，视网膜发育不良与发生密切相关外部限制膜（ELM）中的断裂，这是一个粘合剂/紧密连接的网络根尖过程和感光体内部段。在其他与发育不良相关的视网膜疾病，例如由于突变而引起 Crumbs1（CRB1）和RP27，以及糖尿病性视网膜病。我们已经确定了一种遗传修饰符，该修饰符可防止两种RD7小鼠中榆树的碎片和NRL KO鼠标（RP27）模型。这一发现强烈暗示了榆树的原因光感受器发育不良的发展中的破碎，并提供了一种建立该工具以建立该工具榆树断裂发生的机制，如何导致残留发育不良以及这如何影响疾病进展。为了确定这些机制，我们将：1）确定ELM交界处的自然历史使用细胞连接的标记分析WT和RD7模型中的形成和光感受器分化蛋白质。 2）通过单细胞RNASEQ分析确定ELM碎片涉及的细胞类型。 3）确定修饰蛋白如何改变连接蛋白向细胞膜募集。 4）确定我们生成的其他遗传修饰菌株的分子基础，以进一步了解 NR2E3疾病途径。