A broadly applicable, regulatable strategy to control signaling in the retina.

一种广泛适用的、可调节的策略来控制视网膜信号传导。

• 批准号: 9373910
• 负责人: John Douglas Hulleman
• 金额: $ 24.3万
• 依托单位: UT SOUTHWESTERN MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-30 至 2019-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9373910
• 关键词: Affect; Age; Age related macular degeneration; Antibiotics; Atrophic; Biological Models; Cells; Cellular Stress; Chimeric Proteins; Data; Dependovirus; Development; Dihydrofolate Reductase; Disease; Dose; Eye; Eye diseases; Fundus; Genes; Genetic; Goals; Heat shock proteins; Image; Immunohistochemistry; Individual; Inherited; Injectable; Injection of therapeutic agent; Kinetics; Modeling; Mus; Mutation; Organism; Outcome; Oxidative Stress; Pathway interactions; Phenotype; Physiological; Proteins; Quality Control; Regulation; Research; Research Project Grants; Retina; Retinal; Retinal Degeneration; Retinal Diseases; Signal Pathway; Signal Transduction; Stress; Structure of retinal pigment epithelium; Technology; Tertiary Protein Structure; Testing; Therapeutic; Tissues; Trimethoprim; Western Blotting; Work; age related; aged; base; biological adaptation to stress; cell type; early onset; fluorescence imaging; in vivo; interest; minimally invasive; mouse model; prevent; red fluorescent protein; response; small molecule; stress protein; transcription factor

PROJECT SUMMARY A number of prevalent and currently incurable eye diseases are caused by aberrant oxidative and/or unfolded protein stress. In healthy individuals, these stresses are mitigated by the activation of stress-responsive signaling pathways such as the oxidative stress response and the unfolded protein response. After the cellular stress has been remedied, signaling through these pathways returns to basal levels. However, as an individual ages, or in the case of specific inherited mutations, the ability to effectively recognize stress and activate the appropriate signaling pathways declines5, in many instances resulting in prevalent diseases such as dry age-related macular degeneration (AMD). Thus, one approach to treat stress- related ocular diseases such as AMD would be to restore the appropriate stress-responsive signaling pathways in the eye. However, the technical capability to re-establish this signaling in a physiological context (i.e., conditional sinusoidal expression) in the eye is lacking. Therefore, there is an urgent need to develop a conditional and reversible therapeutic strategy that allows experimental control of the timing and extent of expression of a gene of interest (i.e., a stress- responsive transcription factor) in the eye. Towards this end, in this exploratory/developmental project, we will use adeno-associated virus (AAV) to introduce a small molecule-regulated destabilized domain (DD) strategy to conditionally control protein abundance using mice as a model system. The expected outcomes of this R21 project are to i) validate the use of DD technology for controlling protein levels in the eye in aged mice and ii) to test whether DDs can be used in retinal cells already undergoing degeneration/atrophy. These findings will be critical to determine whether this promising, and potentially broadly applicable DD strategy can be adapted to future research projects targeted at modulating or restoring stress-responsive signaling pathways in the eye. Therefore, successful completion of this project will ultimately positively impact the treatment of currently incurable age-related and inherited eye diseases.

项目概要 许多流行且目前无法治愈的眼部疾病是由异常氧化引起的 和/或未折叠蛋白质应激。在健康的个体中，这些压力可以通过 应激反应信号通路的激活，例如氧化应激反应和 未折叠的蛋白质反应。细胞应激得到缓解后，通过信号传导 这些途径返回到基础水平。然而，随着个人年龄的增长，或者在 特定的遗传突变，有效识别压力并激活压力的能力 适当的信号传导途径下降5，在许多情况下导致流行疾病 例如干性年龄相关性黄斑变性（AMD）。因此，治疗压力的一种方法是 相关眼部疾病如 AMD 将恢复适当的应激反应 眼睛中的信号通路。然而，重建此信令的技术能力 眼睛中缺乏生理背景（即条件正弦曲线表达）。所以， 迫切需要开发一种有条件且可逆的治疗策略，使 实验控制感兴趣基因的表达时间和程度（即应激 眼睛中的反应性转录因子）。为此，在这个探索性/发展性的 项目中，我们将利用腺相关病毒（AAV）引入一种小分子调控的 使用小鼠作为条件控制蛋白质丰度的不稳定结构域（DD）策略 模型系统。该 R21 项目的预期成果是 i) 验证 DD 的使用 控制老年小鼠眼内蛋白质水平的技术，以及 ii) 测试 DD 是否可以 可用于已经发生变性/萎缩的视网膜细胞。这些发现至关重要 以确定这种有前途且可能广泛适用的 DD 策略是否可以 适应未来旨在调节或恢复压力反应的研究项目 眼睛中的信号通路。因此，本项目的顺利完成最终将 对目前无法治愈的与年龄有关的遗传性眼病的治疗产生积极影响。