Do Changes in Thiol Metabolism Mediate Osteoarthritis Progression?

硫醇代谢的变化是否会介导骨关节炎的进展？

• 批准号: 9788031
• 负责人: Mitchell Carl Coleman
• 金额: $ 24.9万
• 依托单位: UNIVERSITY OF IOWA
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-14 至 2021-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9788031
• 关键词: Acetylcysteine; Age; Anatomy; Animal Model; Animals; Antioxidants; Biology; Biomedical Research; Buffers; Cartilage; Cell Nucleus; Cells; Cellular biology; Cellularity; Chondrocytes; Chronic; Clinical; Clinical Research; Data; Degenerative polyarthritis; Dependovirus; Deterioration; Development; Disease; Environment; Enzymes; Etiology; Excision; Family suidae; Fracture; Funding; Gene Targeting; Genes; Glutamate-Cysteine Ligase; Glutathione; Goals; Heme Group; Homeostasis; Hour; Inflammation; Inflammatory; Injury; Intra-Articular Injections; Iowa; Joints; K-Series Research Career Programs; Knockout Mice; Knowledge; Lesion; Lipid Peroxidation; Lipid Peroxides; Lipids; Measures; Mechanics; Mediating; Mediation; Membrane Lipids; Mentors; Mentorship; Metabolic; Metabolic dysfunction; Metabolism; Miniature Swine; Mitochondria; Modeling; Operative Surgical Procedures; Orthopedics; Oryctolagus cuniculus; Outcome; Oxidation-Reduction; Oxidative Stress; Pathogenesis; Pathology; Pathway interactions; Patients; Peroxidases; Pharmacologic Substance; Phase; Positioning Attribute; Prevention; Process; Proteins; Proteoglycan; Protons; Rehabilitation therapy; Reproducibility; Research; Research Personnel; Research Training; Resistance; Role; Scientist; Signal Transduction; Signaling Molecule; Site; Specificity; Structure; Sulfhydryl Compounds; Surgeon; System; TXN gene; Testing; Time; Tissues; Training; Transgenic Mice; Trauma; Traumatic injury; United States National Institutes of Health; Universities; Viral; career; career development; clinically relevant; design; effective therapy; experience; genetic manipulation; glutathione peroxidase; heme oxygenase-1; joint inflammation; joint injury; mitochondrial autophagy; mitochondrial metabolism; novel; operation; outcome forecast; overexpression; oxidation; peroxiredoxin; preservation; prevent; response; response to injury; sample fixation; skills; small molecule; symposium; tenure track; therapeutic target; tool; transcription factor

This career development project is focused on orthopedic training and conducting novel research into the role of thiol metabolism during chronic oxidative stress after mechanical injury to establish an independent line of research. This will be done under the guidance of: primary mentor, Dr. James Martin, an expert in orthopedic cell biology with over 30 years’ experience in research and training; co-mentor for redox biology, Dr. Douglas Spitz, an expert in redox biology with 30 years of continuous independent NIH funding and training experience including an NIH T32 and several K-award mentorships; and clinical co-mentor, Dr. Larry Marsh, a world class trauma surgeon and clinical researcher with experience training clinical and basic scientists. These mentors will guide the acquisition of preliminary data to characterize chondrocyte thiol metabolism and redox signaling, the search and negotiation for a tenure track position, the execution of the R00 research into thiol metabolism after injury, and final R01 composition to establish independence. The training goal of the K99 phase of this project is to provide critical knowledge and experience for operation as an independent orthopedic researcher via incorporation into resident training. Resident conferences will be attended including no fewer than three days a week, 1-2 hours each day. The distinguished University of Iowa Department of Orthopedics and Rehabilitation provides a powerful research environment for pursuit of research goals during the K99 phase, highlighted by large animal models of orthopedic disease. Concurrent with their funded projects and using the same animals, the redox status of intracellular thiols including glutathione and mitochondrial thioredoxin/peroxiredoxin, NRF2/Bach-1 interplay, GPx4 content, and mitochondrial redox function during disease development after intraarticular fracture will be measured. These endpoints have not been explored as a unified pathway, and have not been described in these disease settings. Observing these pathways over three different time points corresponding to early, intermediate, and late stages of disease in a large animal model will allow for a comprehensive understanding of the status of intracellular thiols over time. Taken together with the clinical training described, this will result in an enhanced orthopedic skill set, and development of novel research ideas in well characterized orthopedic models. Detailed description of thiol redox responses during disease will enable logical design of an adeno-associated virus seeking to disrupt thiol metabolism similar to chronic orthopedic disease processes. As the R00 phase commences, data from the K99 phase will guide construction and characterization of the viral tool described. Once this tool has been validated, it will be applied in a rabbit model of post-traumatic injury and complimented with a transgenic mouse resistant to oxidative stress. By comparing responses to injury from control rabbits to rabbits overexpressing Bach-1, a mechanistic role for thiol metabolism in response to inflammation will be described.

这个职业发展项目的重点是骨科培训和进行新的研究， 巯基代谢在机械损伤后慢性氧化应激过程中的作用，以建立一个独立的 研究路线。这将在以下人员的指导下完成：主要导师，詹姆斯·马丁博士， 骨科细胞生物学，拥有30多年的研究和培训经验;氧化还原生物学的共同导师， 博士道格拉斯斯皮茨，氧化还原生物学专家，30年来连续独立的国家卫生研究院的资金， 培训经验，包括NIH T32和几个K奖导师;和临床共同导师，博士。 Larry Marsh是世界级的创伤外科医生和临床研究人员，具有培训临床和 基础科学家这些导师将指导初步数据的采集，以表征软骨细胞 巯基代谢和氧化还原信号，寻找和谈判的任期轨道的位置，执行 研究R 00在损伤后的巯基代谢，并最终确定R 01的组成独立性。 本项目K99阶段的培训目标是为以下人员提供关键知识和经验： 通过纳入住院医师培训，作为独立的骨科研究人员开展手术。居民 参加会议的时间不少于每周三天，每天1-2小时。的 著名的爱荷华州大学骨科和康复系提供了一个强大的 在K99阶段追求研究目标的研究环境，以大型动物为重点 骨科疾病的模型。与他们资助的项目同时进行，使用相同的动物， 细胞内巯基的状态，包括谷胱甘肽和线粒体硫氧还蛋白/过氧化物氧还蛋白，NRF 2/Bach-1 关节内注射后疾病发展过程中的相互作用、GPx 4含量和线粒体氧化还原功能 将测量骨折。这些终点尚未作为统一途径进行探索， 在这些疾病中被描述。在三个不同的时间点观察这些通路 在大型动物模型中，对应于疾病的早期、中期和晚期阶段， 全面了解细胞内硫醇随时间的状态。结合临床 培训描述，这将导致增强骨科技能，并开发新的研究 在良好表征的骨科模型中的想法。疾病期间硫醇氧化还原反应的详细描述 将使得能够合理设计腺相关病毒，寻求破坏类似于 慢性骨科疾病过程。随着R 00阶段的开始，K99阶段的数据将指导 所述病毒工具的构建和表征。一旦该工具得到验证， 应用于创伤后损伤的兔子模型，并与抗 氧化应激通过比较对照兔和过表达Bach-1的兔对损伤的反应， 将描述硫醇代谢在炎症反应中的机制作用。