COLLAGEN CROSS-LINKING IN SKELETAL AGING AND DISEASE

骨骼老化和疾病中的胶原蛋白交联

• 批准号: 7107318
• 负责人: David R Eyre
• 金额: $ 33.41万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 1986
• 资助国家: 美国
• 起止时间: 1986-07-01 至 2010-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7107318
• 关键词: aging; amine oxidoreductase; articular cartilage; biomarker; clinical research; collagen; crosslink; extracellular matrix; histopathology; human tissue; mass spectrometry; molecular pathology; osteoarthritis; osteogenesis imperfecta; osteoporosis; patient oriented research; posttranslational modifications; protein isoforms; protein protein interaction; protein structure function; skeletal system; tissue /cell culture

DESCRIPTION (provided by applicant): The goal is to define in precise detail at the protein level how the lysyl oxidase-mediated cross-linking mechanisms of the major skeletal tissue collagens differ. Insights and questions from studying heritable disorders make it clear that we do not yet understand the local features of collagen cross-linking chemistry and their underlying control mechanisms well enough to relate form to function. It is also clear that the posttranslational quality of collagen can change with age and disease in ways that affect the basic pattern of enzyme-regulated cross-linking. There is growing evidence that such differences between individuals are a risk factor for osteoporosis, osteoarthritis and other degenerative skeletal disorders. The experimental plan targets the biochemistry of collagen cross-linking using advanced mass spectroscopic methods and custom antibodies against cross-linking domains. In parallel, the insights and molecular tools gained from tissue studies will be applied to advance knowledge on existing biomarker assays based on collagen cross-links and develop new biomarkers for monitoring bone and joint disorders non-invasively. The specific aims include: 1) defining the detailed post-translational chemistry of bone collagen including chain usage in the placement and chemical forms of pyridinoline, pyrrole and keto-imine cross-links and differences that occur with age in osteoporosis, osteogenesis imperfecta and other conditions of altered metabolism; 2) compare in similar detail the cross-linking properties of non-mineralized skeletal collagens of cartilages, tendon, ligament and joint capsule; 3) determine the cross-linking properties of type III collagen as a modifier of the extracellular matrix of cartilage and other non-mineralized skeletal tissues; 4) improving and, developing clinically useful biomarkers from the insights gained on collagen cross-linking mechanisms from the tissue studies. The emphasis in the research design and methods is to apply mass spectrometry and other advanced methods in protein analysis to determine details of tissue collagen structure, then use cell culture to explore candidate control mechanisms responsible for the diversity in tissue-specific cross-linking and the changes that occur in aging and disease.

描述（由申请方提供）：目的是在蛋白质水平上精确详细地定义赖氨酰氧化酶介导的主要骨骼组织胶原交联机制的差异。研究遗传性疾病的见解和问题清楚地表明，我们还没有足够好地了解胶原交联化学的局部特征及其潜在的控制机制，以将形式与功能联系起来。同样清楚的是，胶原蛋白的翻译后质量可以随着年龄和疾病而改变，从而影响酶调节的交联的基本模式。越来越多的证据表明，个体之间的这种差异是骨质疏松症、骨关节炎和其他退行性骨骼疾病的风险因素。实验计划的目标是胶原蛋白交联的生物化学，使用先进的质谱方法和针对交联结构域的定制抗体。与此同时，从组织研究中获得的见解和分子工具将被应用于推进基于胶原交联的现有生物标志物测定的知识，并开发用于非侵入性监测骨和关节疾病的新生物标志物。 具体目标包括：1）定义骨胶原的详细翻译后化学，包括吡啶啉、吡咯和酮亚胺交联的位置和化学形式中的链使用，以及在骨质疏松症、骨生成异常和其他代谢改变的情况中随年龄发生的差异; 2）类似详细地比较软骨、肌腱、韧带和关节囊的非矿化骨骼胶原的交联性质; 3）确定作为软骨和其他非矿化骨骼组织的细胞外基质的改性剂的III型胶原的交联性质; 4）从组织研究中获得的胶原交联机制的见解中改进和开发临床上有用的生物标志物。研究设计和方法的重点是应用质谱和其他先进的蛋白质分析方法来确定组织胶原蛋白结构的细节，然后使用细胞培养来探索负责组织特异性交联多样性以及衰老和疾病中发生的变化的候选控制机制。