Collagen Glycation in Aging

衰老中的胶原蛋白糖化

• 批准号: 10225639
• 负责人: David Mark Hudson
• 金额: $ 19.44万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-08-01 至 2024-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10225639
• 关键词: Address; Advanced Glycosylation End Products; Adverse effects; Affect; Age; Aging; Aldehydes; American; Amines; Arginine; Biomechanics; Cartoons; Chemistry; Collagen; Collagen Type I; Connective Tissue; Connective and Soft Tissue; Coupled; Data; Detection; Diabetes Mellitus; Diabetic mouse; Diamond; Disease; Exhibits; Fibrillar Collagen; Fluorescence Spectroscopy; Glucose; Goals; High Pressure Liquid Chromatography; Human; Hydroxylysine; Hyperglycemia; Impaired healing; Individual; Injury; Knowledge; Laboratories; Length; Link; Lysine; Mass Fragmentography; Mass Spectrum Analysis; Measures; Mechanics; Mediating; Methods; Modification; Musculoskeletal; N(6)-carboxymethyllysine; Nature; Non-Insulin-Dependent Diabetes Mellitus; Pathogenicity; Pathologic; Pathology; Pathway interactions; Pattern; Physiological; Population; Prediabetes syndrome; Process; Property; Protein-Lysine 6-Oxidase; Proteins; Publishing; Risk; Sampling; Side; Silicon Dioxide; Site; Skin; Tendon structure; Testing; Therapeutic Uses; Tissues; achilles tendon; adduct; age effect; age related; baby boomer; base; crosslink; glycation; healing; human tissue; innovation; joint mobilization; molecular marker; molecular site; negative affect; novel; pentosidine; prevent; senescence; sugar; targeted treatment; therapeutic target; viscoelasticity

Project Summary/Abstract: In this study, we focus on non-enzymatic glycations (abnormal sugar additions) on collagens in aging tissue. Among the many known cross-links in fibrillar collagens, perhaps the least understood but most speculative pathologically are advanced glycation end-products (AGEs). Normal enzymatic lysyl oxidase-mediated cross- links between individual collagen molecules in a fibril are essential for the strength and integrity of tendons and most musculoskeletal tissues. On the other hand, non-enzymatic glycations are thought to accumulate randomly on aging tissue collagens, with the potential to form abnormal intermolecular cross-links. Glycations and subsequent AGE cross-links in collagens have been linked to limited joint mobility, tissue fragility, and diminished healing capacity, all of which are commonly associated with aging and age-related diseases. Collagen glycations, therefore, not only affect the rapidly aging US population, but also one in three Americans already suffering from prediabetes and hyperglycemia. We propose to 1) identify the main sites of non- enzymatic glycations in type I collagen in aging tendon, and 2) characterize changes in the normal lysyl oxidase-mediated collagen cross-linking associated with non-enzymatic glycations in tendon. Although collagen is a frequently cited substrate for non-enzymatic glycation, the lack of experimental data on any molecular sites of collagen glycation, let alone AGE cross-linking, is striking. Instead, total AGE cross-links in collagen have typically been measured from whole tissue hydrolysates. We propose a paradigm shifting hypothesis that glycations occur preferentially, not randomly, at the helical domain cross-linking lysine residues in tendon type I collagen. These newly identified glycated lysine residues on type I collagen will be compared in aging (young vs. old) human Achilles tendons using targeted ion trap mass spectrometry. We next hypothesize that glycations and AGE products can prevent nascent collagen from enzymatically cross-linking to fibrils in the tendon unit. Changes in the divalent and trivalent collagen cross-link profile will be simultaneously quantitated using an innovative, newly adapted silica hydride based chromatographic approach coupled with established mass spectrometric methods. Age-related glycations at the helical domain cross- linking lysines of type I collagen are predicted to weaken tissues by hindering normal cross-linking as tissues age. Biomechanical testing will be used to correlate increased collagen glycations with changes in tendon material properties. It is essential to locate the primary sites of collagen glycation and their subsequent AGE products before real progress can be made in understanding the specific pathogenic consequences of glycation. These sites could then serve as potential molecular markers of pathology and therapeutic targets. The knowledge gained from this study could help develop targeted therapeutics used to prevent the effects of aging and hyperglycemia in musculoskeletal tissue by preventing abnormal AGE products from forming in collagen.

项目摘要/摘要： 在这项研究中，我们重点关注衰老组织中胶原蛋白的非酶糖化（异常糖添加）。 在纤维状胶原蛋白的许多已知交联中，可能是最不为人所知但最具推测性的 病理上是晚期糖基化终产物（AGE）。正常酶促赖氨酰氧化酶介导的交叉 原纤维中各个胶原蛋白分子之间的联系对于肌腱和肌腱的强度和完整性至关重要 大多数肌肉骨骼组织。另一方面，非酶糖化被认为会积累 随机作用于老化组织胶原蛋白，有可能形成异常的分子间交联。糖基化 随后胶原蛋白中的 AGE 交联与关节活动受限、组织脆弱和 治愈能力下降，所有这些通常都与衰老和年龄相关疾病有关。 因此，胶原蛋白糖化不仅影响迅速老龄化的美国人口，而且影响三分之一的美国人 已经患有糖尿病前期和高血糖。我们建议 1）确定非 老化肌腱中 I 型胶原蛋白的酶促糖基化，以及 2) 表征正常赖氨酰的变化 氧化酶介导的胶原蛋白交联与肌腱中的非酶糖基化相关。虽然 胶原蛋白是一种经常被引用的非酶糖基化底物，但缺乏任何相关的实验数据 胶原蛋白糖化的分子位点（更不用说 AGE 交联）是引人注目的。相反，总 AGE 交联 胶原蛋白通常是从整个组织水解物中测量的。我们提出范式转变 假设糖化优先而非随机地发生在螺旋结构域交联赖氨酸残基处 存在于肌腱 I 型胶原蛋白中。将比较这些新鉴定的 I 型胶原蛋白上的糖化赖氨酸残基 使用靶向离子阱质谱分析老化（年轻与年老）人类跟腱。我们接下来 假设糖基化和 AGE 产物可以阻止新生胶原蛋白的酶交联 到肌腱单位中的原纤维。二价和三价胶原交联谱的变化将是 使用创新的、新调整的基于氢化硅的色谱方法同时进行定量 与已建立的质谱方法相结合。螺旋域交叉处与年龄相关的糖基化 据预测，I 型胶原蛋白的连接赖氨酸会阻碍组织正常交联，从而削弱组织 年龄。生物力学测试将用于将胶原蛋白糖化增加与肌腱变化相关联 材料特性。定位胶原蛋白糖化的主要位点及其随后的 AGE 至关重要 在了解具体致病后果方面取得真正进展之前 糖化。这些位点可以作为病理学和治疗靶点的潜在分子标记。 从这项研究中获得的知识可以帮助开发有针对性的治疗方法，用于预防 通过防止异常 AGE 产物在肌肉骨骼组织中形成来预防衰老和高血糖 胶原。

项目成果

Collagen breaks at weak sacrificial bonds taming its mechanoradicals.

胶原蛋白在弱的牺牲键处断裂，从而驯服了其机械自由基。

• 发表时间: 2023-04-12
• 影响因子: 16.6
• 作者: Rennekamp, Benedikt;Karfusehr, Christoph;Kurth, Markus;Ünal, Aysecan;Monego, Debora;Riedmiller, Kai;Gryn'ova, Ganna;Hudson, David M;Gräter, Frauke
• 通讯作者: Gräter, Frauke