Exploring regulatory mechanisms of glyoxalase-1

探索乙二醛酶-1的调控机制

基本信息

批准号：
10646721
负责人：
JACOB M HAUS
金额：
$ 23.4万
依托单位：
UNIVERSITY OF MICHIGAN AT ANN ARBOR
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 2025-02-28
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10646721
关键词：
Acceleration Acetylation Adult Advanced Glycosylation End Products Aging Alanine Amino Acids Antioxidants Attenuated CRISPR/Cas technology Carbon Cell Culture Techniques Cell Line Cell physiology Cells Clustered Regularly Interspaced Short Palindromic Repeats Coupled Data Deacetylase Deacetylation Disease Drug Metabolic Detoxication Enzymes Gene Dosage Generations Genes Glucose Intolerance Health Heterozygote Human Impairment Inflammation Inflammatory Insulin Resistance Knock-out Knowledge Lactoylglutathione Lyase Longevity Lysine Measures Mediating Metabolic Metabolic Diseases Metabolism Mitochondria Modeling Modification Muscle Muscle Fibers Muscle function Mutagenesis Mutate NADH Non-Insulin-Dependent Diabetes Mellitus Nucleotides Obesity Organ Overnutrition Oxidative Stress Peptides Phenotype Phosphorylation Phosphorylation Inhibition Physiological Polyubiquitination Post-Translational Modification Site Post-Translational Protein Processing Process Production Proteins Proteomics Pyruvaldehyde Reactive Oxygen Species Regulation Role SIRT1 gene Sirtuins Skeletal Muscle Stress System Technology Thinness Threonine Tissues Work adduct adult obesity age related attenuation functional outcomes gain of function genome editing glucose metabolism glycation healthspan in silico in vitro Model inhibitor knock-down loss of function mutant nicotinamide phosphoribosyltransferase nicotinamide-beta-riboside novel obesogenic preservation senescence skeletal muscle metabolism stressor targeted treatment therapeutic target translational model young adult

项目摘要

PROJECT ABSTRACT Methylglyoxal (MG) is a potent intracellular glycating agent that forms advanced glycation endproducts. Formed spontaneously from 3-carbon glycolytic intermediates, MG rapidly glycates proteins and nucleotides, damages mitochondria and directly increases reactive oxygen species production; thus inducing a pro-oxidant state and senescent-like condition. MG and the related glyoxalase enzymatic defense system are emerging as critical players in aging and age-related disease processes. Under physiologic conditions MG is rapidly detoxified by glyoxalase 1 (GLO1). However, when GLO1 is attenuated, MG flux is increased and MG-modified proteins accumulate (termed dicarbonyl stress), both within and outside the cell. Dicarbonyl stress promotes glucose intolerance, oxidative stress and inflammation. The mechanisms regulating GLO1 protein stability and enzymatic activity in skeletal muscle tissue, a tissue critical to glucose metabolism, are not well studied and there is a critical need to understand the functional consequences of reduced GLO1 in the context of obesity, aging and age- related disease. GLO1 is critical to cellular function and subject to numerous posttranslational modifications (PTMs) that regulate GLO1 protein stability and activity. Our objective is to establish robust translational models to delineate the mechanisms by which GLO1 is regulated to better understand the functional consequences of attenuated GLO1. The generation of new, state-of-the-art translational models will help to accelerate the understanding of GLO1 attenuation and dicarbonyl stress and the implications for skeletal muscle health across both the life-span and health-span. We aim to establish the functional relevance of both GLO1 loss and the impact of PTMs of GLO1 in human myotubes. Our approach is to attenuate GLO1 and mutate critical amino acid residues using CRISPR gene editing technology, coupled with measures of dicarbonyl stress. We expect to identify a novel, muscle specific mechanism of GLO1 dysregulation and methylglyoxal-mediated damage. The successful completion of this work will have an important positive impact on advancing the understanding, and provide potential therapeutic targets, to maintain skeletal muscle function with aging and age-related disease.

项目摘要甲基糖（MG）是形成晚期糖基化终产物的有效细胞内糖化剂。形成从3-碳糖水解中间体的自发自发，MG迅速降低蛋白质和核苷酸，损害线粒体并直接增加活性氧的产生；从而诱导促氧化态和类似衰老的状况。 MG和相关的乙二醛酶酶促防御系统正在成为关键衰老和与年龄相关的疾病过程的参与者。在生理条件下，mg通过糖酶1（GLO1）。但是，当glo1衰减时，Mg通量增加并MG修饰蛋白在细胞内外，累积（称为二骨应力）。双骨应力促进葡萄糖不耐受，氧化应激和炎症。调节GLO1蛋白稳定性和酶促的机制在骨骼肌组织中的活性，一种至关重要的葡萄糖代谢的组织，没有很好地研究，并且存在关键需要了解在肥胖，衰老和年龄的背景下降低GLO1的功能后果相关疾病。 GLO1对细胞功能至关重要，受到众多翻译后修饰的影响（PTMS）调节GLO1蛋白稳定性和活性。我们的目标是建立强大的翻译模型描述对GLO1进行调节以更好地理解功能后果的机制减弱GLO1。新的，最先进的翻译模型的产生将有助于加速了解GLO1衰减和双骨压力以及对整个骨骼肌肉健康的影响生命跨度和健康跨度。我们旨在建立GLO1损失和 GLO1的PTM在人肌管中的影响。我们的方法是减弱glo1和突变临界氨基酸使用CRISPR基因编辑技术的残留物，再加上双骨应力的措施。我们希望确定一种新型的GLO1失调和甲基甘氨酸介导的损伤的新型肌肉特异性机制。这成功完成这项工作将对促进理解和提供潜在的治疗靶标，以维持骨骼肌功能，并具有与年龄相关的疾病。