Role of Glucose metabolism in Chondrocyte Mechanotransduction

葡萄糖代谢在软骨细胞力转导中的作用

• 批准号: 9924448
• 负责人: Ronald Kent June
• 金额: $ 34.48万
• 依托单位: MONTANA STATE UNIVERSITY - BOZEMAN
• 依托单位国家: 美国
• 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/9924448
• 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

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.

所有细胞都会受到机械力的影响，并对其做出反应，比如压缩。然而，分子机制 将机制与生物反应联系起来还没有完全理解。我们模型系统的细胞， 软骨细胞在体内受到压缩，这些细胞可以将压缩转化为 生物信号。有证据表明，软骨细胞中葡萄糖的利用受压缩和 生理压力刺激糖酵解，这是软骨细胞制造三磷酸腺苷的主要途径。这 这一现象被认为与软骨细胞维持软骨的能力有关。因此，对葡萄糖的研究 新陈代谢与NIH有关，因为数百万人患有软骨细胞驱动的软骨退化 骨性关节炎。目前的骨关节炎治疗涉及关节运动，这是违反直觉的。我们的节目是为 这是第一次生理相关的培养条件能够在体外压缩软骨细胞。这 项目测试的假设是，正常和骨关节炎软骨细胞的生理性压缩 导致葡萄糖代谢中的特定代谢物模式，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，这 该项目将开发使用机械加载来生产软骨修复的积木的策略。 目标1-体外实验将检查碳(葡萄糖或谷氨酰胺)的来源和机制 监管。因变量包括性别、供者年龄和施压水平(低或高)。 靶向代谢组学数据将从正常和骨关节炎软骨细胞中产生 不同实验条件下的压缩性能。目标2-使用自愿参加的小鼠进行实验 跑步将评估体内的机械转导。因变量包括性别和跑步时间。 读数将包括靶向代谢物和免疫组织学标志物，以检查对 葡萄糖代谢。化验将使用高度特异的酶抑制剂，从而实现逐步分析。 关键的新陈代谢途径。这个项目有很大的创新，包括一个新的系统生物学模型。 以及计算葡萄糖代谢每一步的相对反应速度的分析方法。这些 建模结果将用于改进现有假设和生成新的假设。这样做的目的是 该项目是为了确定正常和正常情况下受压导致的小代谢物模式的变化 骨关节炎软骨细胞。预期的结果是确定候选目标反应 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。 了解这些机制可能有助于开发修复软骨的翻译策略 激活现有的机械敏感通路。深入了解软骨细胞对压缩的反应 通过为软骨修复和使能提供新的治疗靶点来促进骨关节炎的翻译 在临床上取得了实质性进展。