CIS-REGULATORY MECHANISMS UNDERLYING RETINOPATHY

视网膜病变的 CIS 调节机制

• 批准号: 9462155
• 负责人: SHIMING CHEN
• 金额: $ 34.31万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-04-01 至 2021-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9462155
• 关键词: Affect; Age of Onset; Binding; Binding Sites; Biological Assay; ChIP-seq; Chromatin; Computer Simulation; DNA; DNA-Protein Interaction; Data; Development; Disease; Gene Expression; Gene Expression Regulation; Gene Targeting; Genes; Genetic; Genome; Genomic Library; Genomic medicine; Genomics; Global Change; Goals; Human; Knock-in; Knock-in Mouse; Lead; Leber' s amaurosis; Link; Maintenance; Measurement; Measures; Methods; Modeling; Molecular; Mus; Mutation; Pathogenesis; Pathologic; Pathology; Pattern; Phenotype; Photoreceptors; Proteins; Reporter; Reporter Genes; Retina; Retinal Diseases; Retinitis Pigmentosa; Role; Severities; Site; Technology; Testing; Training; base; design; disease-causing mutation; genome-wide; in vivo; individualized medicine; mouse model; mutant; predictive modeling; protein protein interaction; targeted treatment; therapy development; transcription factor

Project Summary    Different human mutations in the photoreceptor-specific gene Crx are linked with multiple retinopathies that vary in their severity and age of onset. How different mutations in this single gene lead to different pathologies is not well understood. Because CRX is a transcription factor, these disease mutations must act by altering the ability of CRX to regulate gene expression. Our goal is to understand how different classes of CRX mutations modify its regulatory function. To achieve this, we will apply a recently developed massively parallel reporter gene technology to systematically and comprehensively measure CRX regulatory function in live retina from multiple mouse knock-in models that carry human CRX disease mutations. We will use the data to train and test a mechanistic, quantitative model that describes how CRX mutations alter protein-DNA and protein-protein interactions to modify gene regulation. In a complementary approach, we will use the knock-in mouse models to determine the effects of CRX disease mutations on its in vivo genome-wide binding, and on the binding of the cooperatively interacting transcription factors OTX2, NRL, and NR2E3. Using these two approaches, we aim to understand how different classes of CRX disease mutations modify gene regulation in photoreceptors, and discover new mechanisms of disease pathogenesis. Our results will provide a strong basis for classifying new CRX mutations, and for designing targeted therapies to treat different genetic forms of retinopathy.

项目总结： -- 人类光感受器特异基因CRX的不同突变与 多种视网膜病变，其严重程度和发病年龄各不相同。有多么不同 这一单一基因的突变导致了不同的病理，目前还不清楚。 因为CRX是一种转录因子，这些疾病突变必须通过改变 CRX调控基因表达的能力。我们的目标是了解 CRX突变的种类改变了它的调节功能。要实现这一点，我们将应用 最近发展起来的大规模平行报告基因技术，以系统和 综合测定多只小鼠活体视网膜CRX的调节功能 携带人类CRX疾病突变的敲入模型。我们将使用这些数据进行训练 并测试一个描述CRX突变如何改变的机械性、量化模型 蛋白质-DNA和蛋白质-蛋白质相互作用，以改变基因调控。在一个 补充方法，我们将使用敲入小鼠模型来确定 CRX病突变对其体内全基因组结合的影响 协同相互作用的转录因子OTX2、NR1和NR2E3的结合。 使用这两种方法，我们的目标是了解不同类别的CRX 疾病突变改变光感受器的基因调控，并发现新的 疾病的发病机制。我们的结果将为以下方面提供强有力的基础 对新的CRX突变进行分类，并设计靶向疗法来治疗不同的 遗传性视网膜病变。