Molecular mechanism underlying late-onset retinal/macular degeneration

迟发性视网膜/黄斑变性的分子机制

基本信息

批准号：
10058720
负责人：
Radha Ayyagari
金额：
$ 46.65万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN DIEGO
依托单位国家：
美国
项目类别：
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/10058720
关键词：
ATAC-seq Ablation Aging Architecture Atlases Biological Assay Biological Models Blindness Bruch&apos s basal membrane structure Cell Line Cell Nucleus Cells Cellular Stress Choroid Chromatin Chronic Complement 1q Complex Dark Adaptation Data Deposition Development Disease Disease Progression Doyne honeycomb retinal dystrophy Drusen Enhancers Exposure to Extracellular Matrix Gene Expression Genes Genomics Goals Human Individual Joints Knock-in Knock-in Mouse Late-Onset Disorder Macular degeneration Mendelian disorder Methods Mitotic Modeling Molecular Mus Mutation Onset of illness Outcome Outcome Study Pathology Pathway interactions Patients Phenotype Physiological Play Proteins Proteome Proteomics Regulator Genes Resolution Retina Retinal Degeneration Role Signal Transduction Small Nuclear RNA Sorsby&apos s fundus dystrophy TIMP3 gene TNF gene Testing Time Tissues Validation Vertebrate Photoreceptors Visual Acuity base cell type clinical phenotype epigenome epigenomics genome editing genomic profiles genomic tools induced pluripotent stem cell innovation member molecular pathology mouse model multimodality neovascularization normal aging novel novel therapeutics therapeutic target transcriptome

项目摘要

ABSTRACT The goal of this proposal is to understand the molecular underpinnings of the well-characterized monogenic disease, Late Onset Retinal/Macular Degeneration (L-ORD/L-ORMD) that recapitulates the major features of other macular degenerations (MDs) with a drusen forming phenotype including AMD although with later onset. L-ORMD is a dominant macular degeneration characterized by the presence of dark adaptation abnormality as early as in the 30s, drusen-like sub-RPE deposits in the 40s, progressive loss of visual acuity, and neovascularization in the 50s leading to irreversible blindness. The dominant Doyne Honeycomb Retinal Dystrophy (DHRD) and Sorsby's Fundus Dystrophy (SFD) as well as the complex disease AMD are examples of other MDs with drusen phenotype. We identified mutations in the gene C1q-TNF-Related Protein 5 (CTRP5/C1QTNF5) in patients with L-ORMD. CTRP5 is secreted by RPE and interacts with EFEMP1 and TIMP3 whose genes have been implicated in DHRD and SFD respectively. All three proteins are components of the extracellular matrix (ECM) and are substrates of the ECM regulator HTRA1. Likewise, AMD associated proteins CFH and C3 are also members of ECM and substrates of HTRA1. These findings support a role for Bruch's membrane (BrM), a specialized ECM of RPE, in MD pathology. S163R Ctrp5 mutation knock-in (KI) mouse models (KI/Wt & KI/KI) that we developed mimic the human L-ORMD phenotype including sub-RPE deposits and BrM abnormalities. We have also established iPSC-RPE of patients with L-ORMD. Using these models, we will (1) characterize the gene regulatory landscape underlying disease pathology by profiling changes in chromatin accessibility and the transcriptome of retinal cells that are the primary and secondary targets of L-ORMD pathology, (2) analyze the proteome profile of BrM-Choroid of these mice to evaluate changes in ECM composition and matricellular proteins with role in signaling associated with aging and with progression of disease to determine the role of ECM in L-ORMD pathology and (3) validate the molecular networks found to play a role in L-ORMD pathology using model systems. The outcomes of this study have the potential to delineate how the molecular networks in each retinal cell type is individually impacted by aging and by progression of disease, and if the retinal cell types adapt to the chronic cellular stress of disease by modulating the epigenome. The studies proposed will significantly enhance our understanding of not only L-ORMD, but also other late-onset pathologies such as AMD.

抽象的该提案的目的是了解特征良好的分子基础单基因疾病，迟发性视网膜/黄斑变性（L-ord/L-ormd），概括了主要其他黄斑变性（MDS）的特征，具有drusen的表型，包括AMD，尽管稍后发作。 L-ormd是一种主要的黄斑变性，其特征是存在暗适应早在30年代，异常，在40年代，视力逐渐丧失，视力丧失， 50年代的新血管形成导致不可逆转的失明。主要的多恩蜂窝视网膜营养不良（DHRD）和SORSBY的眼底营养不良（SFD）以及复杂疾病AMD 其他MD具有drusen表型的示例。我们鉴定了基因C1Q-TNF相关蛋白中的突变 L-ERMD患者的5（CTRP5/C1QTNF5）。 CTRP5由RPE分泌，并与EFEMP1相互作用 Timp3的基因分别与DHRD和SFD有关。这三种蛋白质都是细胞外基质（ECM）的成分，是ECM调节剂HTRA1的底物。同样地， AMD相关的蛋白CFH和C3也是ECM的成员和HTRA1的底物。这些发现在MD病理学中支持BRUCH膜（BRM）的角色，RPE的专业ECM。 S163R CTRP5 突变敲入（Ki）小鼠模型（ki/wt＆ki/ki）我们开发了模拟人类L-ormd 表型，包括子RPE沉积和BRM异常。我们还建立了IPSC-RPE L-ormd患者。使用这些模型，我们将（1）表征基因调节景观潜在的疾病病理学通过分析染色质可及性的变化和视网膜转录组的变化是L-ormd病理学的主要和次要靶标的细胞，（2）分析这些小鼠的BRM-舌形，以评估ECM组成和基质蛋白的变化，并在与衰老和疾病进展相关的信号传导，以确定ECM在L-ormd中的作用病理学和（3）使用模型验证发现在L-ormd病理中起作用的分子网络系统。这项研究的结果有可能描绘每个分子网络如何视网膜细胞类型会受到衰老和疾病进展的单独影响，以及视网膜细胞类型通过调节表观基因组来适应疾病的慢性细胞应激。提出的研究将显着增强了我们对L-ormd的理解，还可以增强其他晚期病理 AMD。