Dermatan sulfate and its role in tissue repair

描述（由申请人提供）：GAG已成为一系列细胞信号传导事件中的关键信号传导分子。尽管硫酸皮肤素（DS）是皮肤和伤口液中含量最丰富的GAG，但这代表了伤口愈合的一个领域，仍有待通过评估修复所需的细胞信号传导事件进行更广泛的评价。DS通过成纤维细胞生长因子-2（FGF-2）和FGF-7介导的信号传导刺激细胞增殖，其作用强于更广泛研究的GAG、硫酸乙酰肝素（HS）。本研究旨在进一步表征DS对体外成纤维细胞生长因子信号传导的影响，并将这些结果外推至人体生理环境。总体假设是FGF-10和FGF-22比HS更喜欢DS，并且伤口环境中的DS结合蛋白的群体可以显著影响组织修复。在目的I中，我们将使用体外生物化学方法确定DS在FGF 10和FGF-22介导的信号传导中是否优于HS。将评估DS依赖性FGF/FGF受体相互作用的结合能力、增殖能力和结构特异性。目的II旨在通过体内分子方法评价人伤口液以鉴定与DS结合比HS更强的FGF。此外，还将评估DS对FGF定位和伤口闭合的影响。拟议的实验将阐明DS在FGF细胞信号传导事件中的作用，这些事件是最佳组织修复所需的，并表征DS结合蛋白的表达模式，这些蛋白最终可用作病理性疾病的诊断标志物。在临床上，已经表明糖胺聚糖在皮肤病的发病机制中起作用。发现DS依赖于必需的FGFR 2-IIIB配体将预测DS合成或降解的障碍将破坏正常的伤口修复。具体而言，我们怀疑异常伤口中存在的DS平衡将不同于正常修复期间观察到的DS平衡，这得到了一些临床研究的支持。因此，分析正常或患病个体的人伤口液中DS的水平和/或硫酸化是本研究提案的前瞻性临床方向，这可能会产生异常伤口愈合的诊断工具和促进正常伤口修复的新治疗方法。总的来说，提出的技术允许暴露于新的分子和生化方法，用其分析蛋白质和碳水化合物的功能和先天免疫在皮肤伤口愈合。

DESCRIPTION (provided by applicant): GAGs have emerged as critical signaling molecules in an array of cell signaling events. Although dermatan sulfate (DS) is the most abundant GAG in the skin and in wound fluid, this represents an area of wound healing which remains to be more extensively evaluated through the assessment of cellular signaling events required for repair. DS has been shown to stimulate cellular proliferation via Fibroblast Growth Factor-2 (FGF-2) and FGF-7 mediated signaling more so than the more extensively studied GAG, heparan sulfate (HS). The purpose of this study is to further characterize the effect of DS on Fibroblast Growth Factor signaling in vitro, and extrapolate these findings to a human physiologic setting. The overall hypothesis is that FGF-10 and FGF-22 prefer DS over HS, and that the population of DS-binding proteins in the wound milieu can significantly influence tissue repair. In Aim I, we will determine if DS is preferred over HS in FGF10 and FGF-22 mediated signaling using an in vitro biochemical approach. The binding capability, proliferative capacity, and structural specificity of DS-dependent FGF/FGF receptor interactions will be assessed. Aim II is designed to evaluate human wound fluid to identify the FGFs that more strongly bind DS over HS through an in vivo molecular approach. In addition, the effect of DS on FGF localization and wound closure will be assessed. The proposed experiments will elucidate the role of DS in FGF-cell signaling events required for optimal tissue repair and characterize the expression patterns of DS-binding proteins, which can ultimately be used as a diagnostic marker in pathologic disease. Clinically, it has been suggested that GAGs play a role in the pathogenesis of skin disease. Finding a dependence for DS by the essential FGFR2-IIIB ligands would predict that a disorder of DS synthesis or degradation would disrupt normal wound repair. Specifically, we suspect that the balance of DS present in abnormal wounds will be different than that observed during normal repair, which is supported by a few clinical studies. Hence, the analysis of the levels and/or sulfation of DS in human wound fluid from normal or diseased individuals is a prospective clinical direction for this research proposal, which may yield a diagnostic tool for abnormal wound healing and novel therapeutic approaches to promote normal wound repair. Overall, the techniques proposed allow for exposure to new molecular and biochemical methods with which to analyze protein and carbohydrate function and innate immunity in dermal wound healing.