Role of Glucose metabolism in Chondrocyte Mechanotransduction

葡萄糖代谢在软骨细胞力转导中的作用

基本信息

批准号：
10400393
负责人：
Ronald Kent June
金额：
$ 12.05万
依托单位：
MONTANA STATE UNIVERSITY - BOZEMAN
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-06-01 至 2024-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10400393
关键词：
Acids Address Affect Age Aging Amino Acids Animals Basic Science Biochemical Reaction Biological Biological Assay Biological Models Biology Body Weight decreased Carbon Cartilage Cartilage Matrix Cells Chondrocytes Citric Acid Cycle Clinical Complex Data Degenerative polyarthritis Deterioration Drug Targeting Elderly Energy-Generating Resources Environment Enzyme Inhibitor Drugs Evaluation Exercise Future Glucose Glutamine Glycolysis Goals Health Histopathology Human Impairment In Vitro Individual Inflammatory Response Injury Isotope Labeling Isotopes Joints Knowledge Link Liquid substance Mass Spectrum Analysis Mechanical Stimulation Mechanics Mediating Metabolic Metabolic Pathway Metabolism Methods Mission Modeling Molecular Motion Movement Mus Musculoskeletal System National Institute of Arthritis and Musculoskeletal and Skin Diseases Non-Essential Amino Acid Outcome Pain Pathogenesis Pathology Pathway interactions Patients Pattern Pentosephosphate Pathway Periodicity Pharmaceutical Preparations Physiological Process Production Proteins Quality of life Reaction Reference Values Regulation Replacement Arthroplasty Respiration Role Running Signal Transduction Source Stains Stimulus Symptoms Synovial Membrane Synovial joint Systems Biology Testing Time Tissues Translating Translations United States National Institutes of Health Walking blood glucose regulation bone bone cell cartilage repair cell injury cell type experimental study glucose metabolism healing improved in vivo inhibitor/antagonist innovation insight joint destruction joint function joint injury joint loading joint mobilization mechanical force mechanical load mechanotransduction metabolomics new therapeutic target novel pre-clinical pre-clinical research reaction rate repaired respiratory response sex small molecule inhibitor translational approach viscoelasticity

项目摘要

All cells are subject to and respond to mechanical forces like compression. However the molecular mechanisms linking the mechanics to biological responses are not fully understood. The cells of our model system, the chondrocytes of cartilage, undergo compression in vivo, and these cells can transduce compression into biological signals. There is evidence that glucose utilization in chondrocytes is regulated by compression and that physiologic compression stimulates glycolysis, the main pathway chondrocytes use to make ATP. This phenomenon has been linked to the ability of chondrocytes to maintain cartilage. Thus, the study of glucose metabolism is relevant to NIH because millions suffer from chondrocyte-driven cartilage deterioration in osteoarthritis. Current osteoarthritis treatments involve joint motion, which is counterintuitive. We show for the first time that physiologically relevant culture conditions enable in vitro compression of chondrocytes. This project tests the hypothesis that physiological compression of both normal and osteoarthritic chondrocytes results in a specific pattern of metabolites within glucose metabolism that support protein production to maintain the cellular microenvironment. The premise is that by quantifying glucose metabolism in chondrocytes this project will develop strategies that use mechanical loading to produce the building blocks for cartilage repair. Aim 1 - In vitro experiments will examine the source of carbon (glucose or glutamine) and the mechanism of regulation. Dependent variables include sex, donor age and the level (low or high) of applied compression. Targeted metabolomics data will be generated from normal and osteoarthritic chondrocytes subjected to compression under different experimental conditions. Aim 2 - Experiments using mice subjected to voluntary running will assess in vivo mechanotransduction. Dependent variables include sex and the duration of running. Readouts will include both targeted metabolites and immunohistological markers examining regulation of glucose metabolism. Assays will employ highly specific enzyme inhibitors that will allow a step-by-step analysis of critical metabolic pathways. This project has substantial innovation including a novel systems biology model and analytical approach that calculate the relative rates of reaction for each step in glucose metabolism. These modeling results will be used both to refine existing hypotheses and to generate new ones. The goal of this project is to identify changes in patterns of small metabolites that result from compression for normal and osteoarthritic chondrocytes. The expected outcome is to identify candidate target reactions that leverage glucose metabolism to increase mechanically driven production of amino acid precursors to repair cartilage. Understanding these mechanisms may prove useful in developing translational strategies to heal cartilage by activating existing mechanosensitive pathways. Insight into how chondrocytes respond to compression will advance osteoarthritis translation by providing new therapeutic targets for cartilage repair and enabling substantial clinical progress.

所有细胞均受到压缩等机械力的影响。但是分子机制将力学与生物反应联系起来尚不完全了解。我们模型系统的单元格软骨软骨细胞，体内受压，这些细胞可以将压缩转换为生物信号。有证据表明，软骨细胞中的葡萄糖利用受压缩和生理压缩刺激糖酵解，这是软骨细胞用来制造ATP的主要途径。这现象与软骨细胞保持软骨的能力有关。因此，葡萄糖的研究代谢与NIH有关，因为数百万人遭受软骨驱动的软骨恶化骨关节炎。当前的骨关节炎治疗涉及关节运动，这是违反直觉的。我们为第一次，生理上相关的培养条件使软骨细胞体外压缩。这项目检验了正常和骨关节炎软骨细胞的生理压缩的假设导致葡萄糖代谢中代谢物的特定模式，该模式支持蛋白质生产以维持细胞微环境。前提是，通过量化软骨细胞中的葡萄糖代谢项目将制定使用机械负载来生产软骨维修的构建块的策略。 AIM 1-体外实验将检查碳的来源（葡萄糖或谷氨酰胺）和机制规定。因变量包括性别，供体年龄和应用压缩的水平（低或高）。靶向代谢组学数据将由正常和骨关节炎软骨细胞产生在不同的实验条件下进行压缩。 AIM 2-使用自愿的小鼠实验跑步将评估体内机械转导。因变量包括性别和跑步持续时间。读数将包括靶向代谢物和免疫组织学标记，以检查调节葡萄糖代谢。测定将采用高度特定的酶抑制剂，以逐步进行分析关键代谢途径。该项目具有实质性的创新，包括新型系统生物学模型和分析方法，这些方法计算葡萄糖代谢中每个步骤的相对反应速率。这些建模结果将既用于完善现有假设并生成新的假设。目标的目标项目是为了确定由正常和骨关节炎软骨细胞。预期的结果是确定利用的候选目标反应葡萄糖代谢增加机械驱动的氨基酸前体的产生以修复软骨。了解这些机制可能对制定翻译策略有用，以治愈软骨激活现有的机械敏感途径。了解软骨细胞对压缩的反应如何通过为软骨修复提供新的治疗靶标，可以提高骨关节炎的翻译实质性的临床进展。