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，我们着重于衰老组织胶​​原蛋白的非酶糖糖性（添加异常糖）。 在纤维状胶原中的许多已知的交联，也许是最不了解的，但最投机性的 病理上是晚期糖基化终产物（年龄）。正常酶赖氨酸氧化酶介导的交叉 原纤维中个体胶原分子之间的联系对于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，非酶糖糖被认为会累积 在衰老的组织胶原蛋白上随机，可能形成异常的分子间交联。糖基 胶原蛋白的随后年龄交联已与关节迁移率有限，组织脆弱性和 愈合能力降低，所有这些通常都与衰老和与年龄有关的疾病有关。 因此，胶原蛋白糖浆不仅会影响美国人口迅速衰老，而且还会影响三分之一的美国人 已经患有糖尿病前和高血糖。我们建议1）确定非 - I型胶原蛋白胶原蛋白蛋白胶原蛋白的酶糖糖浆和2）表征正常赖氨酸的变化 氧化酶介导的胶原蛋白交联与肌腱中的非酶糖糖有关。虽然 胶原蛋白是一种经常引用的非酶糖化底物，缺乏任何实验数据 胶原蛋白酶的分子位点，更不用说年龄的交联，令人震惊。相反，总年龄交联 胶原蛋白通常是从整个组织水解物中测量的。我们提出了一个范式移动 假设糖糖化优先发生，而不是随机发生在螺旋结构域交联赖氨酸残基 在肌腱I型胶原蛋白中。将比较这些新鉴定的I型胶原蛋白糖化赖氨酸残基 在衰老（年轻）中，使用靶向离子陷阱质谱法。我们接下来 假设糖浆和年龄产品可以防止新生的胶原蛋白酶线交联 到肌腱单元中的原纤维。二价和三价胶原蛋白交联轮廓的变化将是 同时使用创新的，新型二氧化硅的色谱方法进行定量 再加上已建立的质谱法。在螺旋结构域交叉的年龄相关糖糖 I型胶原蛋白的赖氨酸连接通过阻碍正常的交叉链接作为组织来削弱组织 年龄。生物力学测试将用于将增加的胶原蛋白糖化与肌腱的变化相关联 材料特性。至关重 可以在了解真正进步之前的产品，以了解 糖化。然后，这些位点可以用作病理和治疗靶标的潜在分子标记。 从这项研究中获得的知识可以帮助开发用于防止影响的有针对性的治疗剂 通过防止异常年龄产物形成肌肉骨骼组织的衰老和高血糖 胶原。

项目成果

Collagen breaks at weak sacrificial bonds taming its mechanoradicals.

• DOI: 10.1038/s41467-023-37726-z
• 发表时间: 2023-04-12
• 期刊: NATURE COMMUNICATIONS
• 影响因子: 16.6
• 作者: Rennekamp, Benedikt;Karfusehr, Christoph;Kurth, Markus;uenal, Aysecan;Monego, Debora;Riedmiller, Kai;Gryn'ova, Ganna;Hudson, David M.;Graeter, Frauke
• 通讯作者: Graeter, Frauke