Characterization of DHDDS RP59 Knockin Models of Retinitis Pigmentosa

视网膜色素变性 DHDDS RP59 敲入模型的表征

• 批准号: 10507785
• 负责人: Mai N Nguyen
• 金额: $ 3.86万
• 依托单位: UNIVERSITY OF ALABAMA AT BIRMINGHAM
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-03-15 至 2026-03-14
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10507785
• 关键词: 1p36; Affect; Alleles; Anatomy; Animal Model; Biochemical; Biochemical Pathway; Blindness; CRISPR/Cas technology; Cells; Chromosome Mapping; Chromosomes; Clinical; Congenital disorders of glycosylation; Data; Diphosphates; Disease; Electroretinography; Exons; Functional disorder; Genes; Glial Fibrillary Acidic Protein; Hereditary Disease; Human; Immunohistochemistry; Individual; Inherited; Intervention; Knock-in; Knock-in Mouse; Lead; Link; Measurement; Missense Mutation; Modeling; Molecular; Morphology; Mus; Mutation; Online Mendelian Inheritance In Man; Pathology; Patients; Phenotype; Photoreceptors; Physiological; Physiology; Plant Roots; Point Mutation; Pre-Clinical Model; Preclinical Testing; Protein Glycosylation; Proteins; Publishing; Quality of life; Recovery; Reporting; Retina; Retinal Degeneration; Retinal Diseases; Retinal Dystrophy; Retinitis Pigmentosa; Severity of illness; Synapses; Synaptic Transmission; Technology; Time; Up-Regulation; base; expectation; glycosylation; inorganic phosphate; insight; light microscopy; loss of function; mouse model; novel; outer plexiform layer; prenyl; response; retinal rods

Project Summary/Abstract Retinitis pigmentosa (RP) is a heterogeneous group of disorders that cause retinal degeneration. To date, mutations in more than 58 genes have been linked to classic retinitis pigmentosa, and collectively all forms of inherited retinal dystrophies involves at least 300 genes. My studies are focused on retinitis pigmentosa-59 (RP59), a non-syndromic autosomal recessive form of RP caused by mutations in the dehydrodolichyl diphosphate synthase (DHDDS) gene, localized to exon 3 on chromosome 1p36.11. The most prevalent mutation is K42E, while T206A and R98W also have been reported. We generated a murine Dhdds K42E knock- in mouse model using CRISPR/Cas9 gene editing technology. Preliminary data shows significant reduction of ERG b-wave amplitudes and we have reported marked GFAP up-regulation, without retinal degeneration or decreased protein N-glycosylation. This suggests that defective DHDDS-dependent retinal degeneration may be more complicated than simply a loss-of-function mechanism due to altered protein glycosylation. To better understand the basis for selective retinal pathology associated with RP59 mutations, we plan to characterize the DhddsK42E/K42E knock-in mouse anatomically and physiologically. Additionally, we will create and analyze two new models of DHDDS-related inherited retinal disease, Dhdds T206A/T206A and DhddsT206A/K42E. Studying the retina in these mouse models will give us insight into the pathophysiology of this inherited disorder, and may lead to novel treatment options for this disorder.

项目概要/摘要 色素性视网膜炎 (RP) 是一组导致视网膜变性的异质性疾病。迄今为止， 超过 58 个基因的突变与经典色素性视网膜炎以及所有形式的视网膜色素变性有关 遗传性视网膜营养不良涉及至少 300 个基因。我的研究重点是色素性视网膜炎-59 (RP59)，一种非综合征性常染色体隐性遗传形式的 RP，由脱氢多利奇基突变引起 二磷酸合酶 (DHDDS) 基因，位于染色体 1p36.11 上的外显子 3。最普遍的 突变为K42E，同时也有报道T206A和R98W。我们生成了小鼠 Dhdds K42E 敲击- 使用 CRISPR/Cas9 基因编辑技术在小鼠模型中进行实验。初步数据显示显着减少 ERG b 波振幅，我们报告了明显的 GFAP 上调，没有视网膜变性或 蛋白质 N-糖基化降低。这表明 DHDDS 依赖性视网膜变性缺陷可能是 比简单地由于蛋白质糖基化改变而导致的功能丧失机制更为复杂。为了更好 了解与 RP59 突变相关的选择性视网膜病理学的基础，我们计划表征 DhddsK42E/K42E 敲入小鼠的解剖学和生理学。此外，我们将创建并分析两个新的 DHDDS 相关遗传性视网膜疾病模型，Dhdds T206A/T206A 和 DhddsT206A/K42E。研究视网膜 这些小鼠模型将使我们深入了解这种遗传性疾病的病理生理学，并可能导致新的发现 这种疾病的治疗选择。