Targeting Nr2e3 to prevent photoreceptor degeneration

靶向 Nr2e3 预防光感受器变性

基本信息

批准号：
10587113
负责人：
JOSEPH CORBO
金额：
$ 52.12万
依托单位：
WASHINGTON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-03-01 至 2028-02-28
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10587113
关键词：
Acute Adult Adverse effects Affect Animal Disease Models Animal Model Behavioral Behavioral Assay Biological Assay Blindness CRISPR/Cas technology Candidate Disease Gene Cells Cessation of life Clinical Research Cone Data Dependovirus Development Disease Disease Progression Disease model Electron Microscopy Electrophysiology (science)Functional disorder Future Gene Expression Gene Expression Profiling Genes Genetic Genetic Heterogeneity Goals Histologic Human Hybrid Cells Immunohistochemistry Knock-out Light Mediating Modeling Molecular Morbidity - disease rate Morphology Motivation Mus Mutation Patients Persons Photoreceptors Physiological Retinal Degeneration Retinal Dystrophy Retinitis Pigmentosa Rod Source Structure Testing Therapeutic Treatment Efficacy Vision Vision research Wild Type Mouse Work behavioral phenotyping candidate selection cell type effective therapy effectiveness testing efficacy evaluation inherited retinal degeneration mouse model neuroprotection new therapeutic target novel novel strategies novel therapeutics overexpression photoreceptor degeneration preservation prevent retinal rods rho targeted treatment transcription factor transcriptome sequencing

项目摘要

Project Summary Retinitis pigmentosa (RP) is the most common form of retinal dystrophy and can be caused by mutations in any one of dozens of rod-enriched genes. The genetic heterogeneity of RP represents a major challenge for the development of effective therapies. For this reason, gene-independent treatments for RP have become a long- sought goal in vision research. In this proposal, we will test the hypothesis that knockout of the rod-specific transcription factor Nr2e3 prevents photoreceptor degeneration in multiple mouse disease models. In Specific Aim 1, we will characterize the neuroprotective effects of developmental Nr2e3 knockout in multiple models of photoreceptor degeneration, including a light-damage model and four mechanistically diverse models of RP (Pde6brd10/rd10, RhoP23H/+, Rho-/- and Cngb1-/-). We will use a combination of molecular, cellular, physiological, and behavioral assays to evaluate the efficacy and versatility of this therapeutic approach. In Specific Aim 2, we will evaluate the therapeutic potential of acute, adeno-associated virus (AAV)-delivered, CRISPR-Cas9-mediated Nr2e3 knockout in the same four mouse RP models used in Aim 1. For each model, we will evaluate the ability of acute Nr2e3 knockout to protect photoreceptors at multiple stages of degeneration. We will also compare the effects of acute Nr2e3 knockout to those of Nr2e3 overexpression, which has also been suggested to prevent degeneration. Together, these studies will test the effectiveness of Nr2e3-based reprogramming as a gene- independent therapy for RP. Finally, in Specific Aim 3, we will determine the effects of acute Nr2e3 knockout in wild-type mouse rods and identify neuroprotective factors downstream of Nr2e3. We will first compare the effects of acute Nr2e3 knockout to those of developmental Nr2e3 knockout. We will then perform RNA-seq on Nr2e3- knockout rods to generate a list of Nr2e3-downstream candidate effector genes. We will knockout or overexpress selected candidate genes, singly and in combination, in four mouse models of RP to identity those that confer a neuroprotective effect. If successful, these studies will establish Nr2e3 knockout as a novel gene-independent therapy for RP, paving the way for future studies in large-animal models of photoreceptor degeneration and for clinical studies in human patients.

项目摘要色素性视网膜炎（RP）是视网膜营养不良的最常见形式，可能是由任何突变引起的数十个富含杆的基因之一。 RP的遗传异质性代表了对开发有效的疗法。因此，对RP的基因无依赖性治疗已成为长期在视力研究中寻求目标。在此提案中，我们将测试杆特异性敲除的假设转录因子NR2E3可防止多种小鼠疾病模型中的光感受器变性。具体 AIM 1，我们将表征发育NR2E3敲除多种模型的神经保护作用光感受器的变性，包括轻伤模型和四个机械多样性的RP模型（pde6brd10/rd10，rhop23h/+，rho - / - 和cngb1 - / - ）。我们将结合分子，细胞，生理和行为分析以评估这种治疗方法的功效和多功能性。在特定的目标2中，我们将评估急性，腺相关病毒（AAV）传递，CRISPR-CAS9介导的治疗潜力在AIM 1中使用的相同四个鼠标RP模型中的NR2E3敲除。对于每个模型，我们将评估能力在多个变性阶段保护光感受器的急性NR2E3敲除。我们还将比较急性NR2E3敲除对NR2E3过表达的影响，这也已被建议防止退化。总之，这些研究将测试基于NR2E3的重编程作为基因的有效性 RP的独立治疗。最后，在特定的目标3中，我们将确定急性NR2E3敲除的影响野生型小鼠杆并鉴定NR2E3下游的神经保护因子。我们将首先比较效果急性NR2E3敲除发展的NR2E3敲除。然后，我们将在NR2E3-上执行RNA-Seq 敲除杆生成NR2E3-Downstream候选效应子基因的列表。我们将淘汰或过表达在RP的四种小鼠模型中，选定的候选基因是赋予A的四种小鼠模型神经保护作用。如果成功，这些研究将建立NR2E3敲除作为一种新型基因独立的 RP的治疗，为在大动物模型的光感受器变性和用于未来的研究铺平了道路人类患者的临床研究。