Writing and erasing O-GlcNAc on target proteins in the brain

在大脑中的目标蛋白上写入和擦除 O-GlcNAc

• 批准号: 10637668
• 负责人: Christina Woo
• 金额: $ 175.63万
• 依托单位: HARVARD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-04-15 至 2026-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10637668
• 关键词: Acceleration; Affect; Alzheimer disease prevention; Alzheimer' s Disease; Alzheimer' s disease brain; Alzheimer' s disease model; Alzheimer' s disease risk; Alzheimer' s disease therapy; Binding; Biological Assay; Biological Models; Biological Process; Brain; Brain region; Cells; Chemicals; Circadian Rhythms; Clinic; Clinical Trials; Complement; Cytoplasm; Cytoplasmic Protein; Data Set; Defect; Distress; Drosophila genus; Drosophila inturned protein; Enzymes; Evaluation; Event; Genes; Genetic study; Glucosamine; Glycoproteins; Goals; Homer protein; In Vitro; Intervention; Investigation; Joints; Knock-in; Letters; Libraries; Link; Maps; Measurement; Measures; Methods; Mitochondria; Mitochondrial Proteins; Modality; Modeling; Modification; Molecular; Neurodegenerative Disorders; Neurons; Nuclear; Nuclear Proteins; Nutrient; Nutrient availability; O-GlcNAc transferase; Organism; Outcome; Parkinson Disease; Pathogenesis; Pathology; Phenotype; Physiological; Physiological Processes; Play; Post-Translational Protein Processing; Prevention; Protein Engineering; Proteins; Proteome; Public Health; Regulation; Research Personnel; Role; Site; Sleep; Sleep Wake Cycle; Sleep disturbances; Symptoms; System; Tauopathies; Writing; behavioral study; cell type; circadian; disease phenotype; drug repurposing; flexibility; fly; glycoproteomics; in vitro Assay; in vivo; inhibitor; innovation; insight; knock-down; nanobodies; novel; peptide O-linked N-acetylglucosamine-beta-N-acetylglucosaminidase; prevent; remediation; sensor; sleep abnormalities; sleep regulation; small molecule; small molecule libraries; sugar; supportive environment; targeted treatment; tau Proteins; therapeutic target; tool

WRITING AND ERASING O-GLCNAC ON TARGET PROTEINS IN THE BRAIN PROJECT SUMMARY O-Linked N-acetyl glucosamine (O-GlcNAc) is a nutrient sensor that dynamically modifies nuclear, cytoplasmic, and mitochondrial proteins. Dysregulation of O-GlcNAc has been linked to disruptions in sleep and circadian rhythm and several neurodegenerative diseases, including Alzheimer’s Disease (AD). While sleep and circadian rhythm defects are distressing symptoms of AD and other tauopathies, sleep disturbance may be a major risk factor for AD and is thought to accelerate its pathology. Extensive studies on the association of O-GlcNAc to AD have led to the first clinical trials targeting O-GlcNAc for therapy. However, these inhibitors globally alter the O- GlcNAc proteome, where a more targeted strategy may provide greater benefit. A systematic investigation of the connection between the O-GlcNAc modification and sleep regulation and AD pathogenesis would significantly impact the discovery of novel mechanisms to provide new avenues for targeted prevention and therapy. O-GlcNAc is regulated by nutrient availability and the complementary activity of two enzymes: O-GlcNAc transferase (OGT) writes the modification and O-GlcNAcase (OGA) erases it from proteins. Recently, innovations in protein engineering and gene editing tools developed by the co-investigators have provided access to precise tuning of O-GlcNAc on specific neurons and desired target proteins in the brain of Drosophila model systems of sleep and AD. Here, we will capitalize on the joint expertise in the Woo Lab and Walker Lab to facilitate the first systematic study to measure, map, and manipulate O-GlcNAc from desired target proteins and in specific neurons in the brain to yield crucial insights to the pathogenesis of AD and novel chemical strategies for remediation. To meet this goal, we will take a three-pronged approach. We will first systematically examine the relationship between O-GlcNAc in specific neurons of Drosophila models of sleep and AD pathogenesis to identify the brain regions that are most dependent on O-GlcNAc regulation. Second, we will use a targeted writer and eraser of O-GlcNAc, developed through protein engineering, to systematically examine the role of O-GlcNAc on selected target proteins in the brain to identify drivers and potential targets for alleviating sleep disruptions and AD pathogenesis. Third, we will pursue the discovery of small molecules that selectively write and erase O-GlcNAc in vitro and in vivo, which will complement our protein engineering approaches and provide targeted alternatives to global inhibitors that are under evaluation for AD therapy in the clinic. The successful outcome of this proposal will afford validated Drosophila models of sleep and AD with neuron-specific manipulation of OGT and OGA or specific target proteins using target writers and erasers of O-GlcNAc, with associated maps of O-GlcNAc proteins and sites, in addition to new and selective small molecule editors of O- GlcNAc to enable more targeted therapeutic approaches in the long-term. Additionally, the systematic methods and tools to connect physiological measurements to molecular function developed here will be translatable to the study of the connection of O-GlcNAc to other neurodegenerative diseases and beyond.

在大脑中的靶蛋白上编写和删除O-GLCNAC 项目摘要 O连接的N-乙酰葡萄糖胺（O-GLCNAC）是一种营养传感器，可以动态修饰核，细胞质， 和线粒体蛋白。 O-GLCNAC的失调与睡眠和昼夜节律的干扰有关 节奏和几种神经退行性疾病，包括阿尔茨海默氏病（AD）。而睡眠和昼夜主义者 节奏缺陷是AD和其他功的令人痛苦的症状，睡眠障碍可能是主要风险 AD的因素，被认为可以加速其病理。关于O-GLCNAC与AD的关联的广泛研究 导致了针对O-GLCNAC治疗的首次临床试验。但是，这些抑制剂在全球范围内改变了O- GlcNAC蛋白质组，更具针对性的策略可能会提供更大的好处。系统投资的 O-GLCNAC修饰与睡眠调节与AD发病机理之间的联系将显着 影响发现新机制的发现，为靶向预防和治疗提供了新的途径。 O-GLCNAC受养分可用性和两种酶的互补活性的调节：O-GlCNAC 转移酶（OGT）写入修饰，O-Glcnacase（OGA）从蛋白质中删除它。最近， 共同研究器开发的蛋白质工程和基因编辑工具的创新已提供 访问果蝇大脑中特定神经元和所需靶蛋白上O-GLCNAC的精确调整 睡眠和广告的模型系统。在这里，我们将利用Woo Lab和Walker Lab的共同专业知识 促进第一个系统的系统研究，以测量，映射和操纵所需靶蛋白的O-GLCNAC 在大脑中的特定神经元中，对AD和新型化学的发病机理产生关键见解 补救策略。为了实现这一目标，我们将采取三管齐下的方法。我们将首先系统地 检查果蝇模型和AD的特定神经元中O-GLCNAC之间的关系 发病机理确定最依赖于O-GLCNAC调节的大脑区域。第二，我们会的 使用通过蛋白质工程开发的O-GLCNAC的有针对性的作家和橡皮擦，以系统地检查 O-GLCNAC在大脑中选定的靶蛋白上的作用，以识别驱动因素和潜在的靶标以减轻 睡眠破坏和AD发病机理。第三，我们将追求有选择性的小分子的发现 在体外和体内写入并擦除O-GlcNAC，这将补充我们的蛋白质工程方法 提供针对诊所AD疗法的全球抑制剂的有针对性替代方法。 该提案的成功结果将提供经过验证的果蝇模型，并具有神经特异性的AD 使用O-GLCNAC的目标作者和橡胶对OGT和OGA或特定目标蛋白操纵 O-GLCNAC蛋白和位点的相关图，除了O-的新的和选择性的小分子编辑器外 从长远来看，GLCNAC可以实现更具针对性的治疗方法。此外，系统方法 以及将物理测量连接到此处开发的分子功能的工具将转换为 O-GLCNAC与其他神经退行性疾病及其他地区的联系的研究。