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

基本信息

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
项目类别：
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&apos s Disease Alzheimer&apos s disease brain Alzheimer&apos s disease model Alzheimer&apos s disease risk Alzheimer&apos 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

项目摘要

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.

在大脑中的目标蛋白上写入和擦除 O-GLCNAC 项目概要 O-连接的 N-乙酰氨基葡萄糖 (O-GlcNAc) 是一种营养传感器，可动态修饰细胞核、细胞质、 O-GlcNAc 的失调与睡眠和昼夜节律紊乱有关。节律和多种神经退行性疾病，包括阿尔茨海默病 (AD) 睡眠和昼夜节律。节律缺陷是 AD 和其他 tau蛋白病的令人痛苦的症状，睡眠障碍可能是主要风险 O-GlcNAc 与 AD 之间的关系被认为可以加速其病理学的广泛研究。首个针对 O-GlcNAc 治疗的临床试验已经开展。然而，这些抑制剂在全球范围内改变了 O-GlcNAc。 GlcNAc 蛋白质组，更有针对性的策略可能会带来更大的益处。 O-GlcNAc 修饰与睡眠调节和 AD 发病机制之间的联系将显着影响新机制的发现，为有针对性的预防和治疗提供新途径。 O-GlcNAc 受营养可用性和两种酶的互补活性调节：O-GlcNAc 最近，转移酶 (OGT) 写入修饰，而 O-GlcNAcase (OGA) 将其从蛋白质中删除。联合研究人员开发的蛋白质工程和基因编辑工具的创新提供了获得对果蝇大脑中特定神经元和所需目标蛋白的 O-GlcNAc 的精确调节在这里，我们将利用 Woo 实验室和 Walker 实验室的联合专业知识。促进首次系统研究从所需目标蛋白中测量、绘制和操作 O-GlcNAc 以及大脑中的特定神经元，以对 AD 的发病机制和新型化学物质产生重要的见解为了实现这一目标，我们将采取三管齐下的方法。检查果蝇睡眠模型特定神经元中的 O-GlcNAc 与 AD 之间的关系其次，我们将确定最依赖 O-GlcNAc 调节的大脑区域。使用通过蛋白质工程开发的 O-GlcNAc 靶向写入器和擦除器来系统地检查 O-GlcNAc 对大脑中选定目标蛋白的作用，以确定缓解的驱动因素和潜在目标第三，我们将致力于发现选择性的小分子。在体外和体内写入和擦除 O-GlcNAc，这将补充我们的蛋白质工程方法和为正在临床评估 AD 治疗的全球抑制剂提供有针对性的替代品。该提案的成功结果将提供经过验证的睡眠和 AD 神经元特异性果蝇模型使用 O-GlcNAc 的目标写入器和擦除器操作 OGT 和 OGA 或特定目标蛋白， O-GlcNAc 蛋白和位点的相关图谱，以及新的选择性 O-小分子编辑器 GlcNAc 能够实现更有针对性的长期治疗方法。这里开发的将生理测量与分子功能测量连接起来的工具将可转化为研究 O-GlcNAc 与其他神经退行性疾病及其他疾病的关系。