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.

项目摘要 许多流行的和目前无法治愈的眼部疾病是由异常的氧化应激引起的。 和/或未折叠蛋白质应激。在健康的个体中，这些压力通过 激活应激反应信号通路，如氧化应激反应， 未折叠的蛋白质反应。在细胞压力得到补救后， 这些通路恢复到基础水平。然而，随着个体年龄的增长，或者在 特定的遗传突变，能够有效地识别压力并激活 适当的信号通路下降5，在许多情况下导致流行疾病 如干性年龄相关性黄斑变性（AMD）。因此，一种治疗压力的方法- 相关的眼部疾病，如AMD将恢复适当的应激反应， 眼睛中的信号通路。然而，在网络中重新建立这种信令的技术能力是有限的。 生理环境（即，条件正弦表达式）在眼睛中缺乏。因此，我们认为， 迫切需要开发一种有条件的和可逆的治疗策略， 对感兴趣基因表达的时间和程度的实验控制（即，一种压力- 反应性转录因子）。为此，在这种探索性/发展性的 项目，我们将使用腺相关病毒（AAV）引入一个小分子调节的 不稳定结构域（DD）策略，以有条件地控制蛋白质丰度，使用小鼠作为 模型系统。该R21项目的预期成果是：i）验证DD的使用 用于控制老年小鼠眼睛中蛋白质水平的技术和ii）测试DD是否可以 用于已经经历变性/萎缩的视网膜细胞。这些发现将至关重要 以确定这一有前途的、可能广泛适用的DD战略是否可以 适应未来的研究项目，旨在调节或恢复应激反应 眼睛中的信号通路。因此，该项目的成功完成将最终 积极影响目前无法治愈的年龄相关和遗传性眼病的治疗。