Deciphering the oxidation status of biomolecules in synovial fluids and tissues acting as damage associated molecular patterns (DAMPs) in rheumatoid arthritis

破译滑液和组织中生物分子的氧化状态，作为类风湿性关节炎损伤相关分子模式 (DAMP)

基本信息

批准号：
264172152
负责人：
Professor Dr. Ralf Hoffmann
金额：
--
依托单位：
Institut für Medizinische Physik und Biophysik
依托单位国家：
德国
项目类别：
Research Grants
财政年份：
2014
资助国家：
德国
起止时间：
2013-12-31 至 2018-12-31
项目状态：
已结题

来源：
https://gepris.dfg.de/gepris/projekt/264172152?language=en
关键词：
Deciphering oxidation status biomolecules synovial

项目摘要

Arthritis, which can be due to degeneration or autoimmunity, is a major cause of disability and early retirement in the industrialized countries. For instance, more than 2 million German citizens were diagnosed to be markedly affected by cartilage degradation and/or loss of function of one or more joints. Thus, the socio-economic consequences and costs for health care are immense. Even worse, the incidence of the disease is continuously increasing.The excessive production of reactive oxygen species (ROS), such as hypochorus acid (HOCl) and hydroxyl radicals, is an important hallmark of inflammatory diseases, especially for rheumatoid arthritis (RA). These highly reactive species have the potential to modify nearly all biomolecules: effects towards proteins, glycosaminoglycans (the prime carbohydrates of the extracellular matrix of cartilage) and the (poly)unsaturated fatty acyl residues within phospholipids could be unequivocally substantiated. However, there is a considerable lack of knowledge about the products that are formed, for example, by oxidatively modified lipids (with reactive aldehyde groups) reacting with the amino groups of abundant proteins or glycosaminoglycans. Additionally, it is increasingly recognized that endogenous molecules modified by ROS (damage-associated molecular patterns; DAMPs) activate cellular receptors that are accompanied by increased inflammation. We hypothesize that these known but "non-studied" substances are particularly relevant for the pathogenesis of inflammatory diseases, i.e. that oxidized compounds are particularly relevant for the pathogenesis of inflammatory rheumatic diseases and are thus of diagnostic and therapeutic importance. We deal with these aspects on different levels of complexity. First, we will synthesize oxidized and chlorinated fatty acids and phospholipids. The detailed characterization of these products by high resolution chromatography, mass spectrometry and NMR spectroscopy is a very challenging task, as methods have to be first adapted to these molecules. Even more challenging will be the analysis of reaction products formed between oxidized fatty acids/lipids and glycosaminoglycans or peptides mimicking the active sites of proteins (lubricin, aggrecan) involved in rheumatic diseases. Second, oxidized molecule classes identified in the first aim will be investigated both in synovial fluid and extracts from the synovial membranes from patients suffering from rheumatic diseases in relation to the severity of the disease. Third, we will judge the relevance of identified oxidized PL, GAG, and proteins as oxidative DAMPs on the cellular immune response. For this task, their effects on T cells, antigen presenting cells and monocytes from the rheumatoid synovium will be studied in vitro by measuring typical inflammation markers such as interferon-gamma and selected cytokines that are released by the cells.

关节炎可能是由于变性或自身免疫性引起的，是工业化国家的残疾和提前退休的主要原因。例如，被诊断出超过200万德国公民会受到软骨降解和/或一个或多个关节功能丧失的明显影响。因此，医疗保健的社会经济后果和成本是巨大的。更糟糕的是，该疾病的发生率正在不断增加。活性氧（ROS）的过量产生，例如酸（HOCL）和羟基自由基，是炎症性疾病的重要标志，尤其是对于类风湿关节炎（RA）。这些高反应性的物种具有修改几乎所有生物分子的潜力：对蛋白质的作用，软骨细胞外基质的主要碳水化合物（糖胺聚糖）和磷脂内（多饱和的）不饱和脂肪酰基残基的作用。然而，存在相当大的知识，例如通过氧化修饰的脂质（具有反应性醛基）与丰富的蛋白质或糖胺聚糖的氨基反应。此外，越来越多地认识到，由ROS（损伤相关的分子模式；抑制作用）修饰的内源性分子激活了伴随着炎症增加的细胞受体。我们假设这些已知但“未研究”的物质与炎症性疾病的发病机理特别相关，即氧化化合物与炎症性风湿性疾病的发病机理特别相关，因此具有诊断性和治疗性的重要性。我们在不同级别的复杂性上处理这些方面。首先，我们将合成氧化和氯化脂肪酸和磷脂。通过高分辨率色谱，质谱和NMR光谱法对这些产品的详细表征是一项非常具有挑战性的任务，因为必须首先适应这些分子。更具挑战性的是分析氧化脂肪酸/脂质与糖胺聚糖或模仿涉及风湿性疾病的蛋白质活性位点的反应产物。其次，在第一个目的中确定的氧化分子类将在滑液中研究和从滑膜膜上的提取物中的提取物，这些患者与疾病严重程度有关。第三，我们将判断已识别的氧化PL，GAG和蛋白质的相关性是细胞免疫反应上的氧化湿度。为此，通过测量细胞释放的典型炎症标志物（例如干扰素 - 伽马市）和精选的细胞因子，将在体外研究它们对T细胞，抗原呈递细胞和单核细胞的影响。