Dermatan sulfate and its role in tissue repair

描述（由申请人提供）：在一系列细胞信号事件中，gag已成为关键的信号分子。虽然皮肤硫酸酯（DS）是皮肤和伤口液中最丰富的GAG，但它代表了伤口愈合的一个领域，需要通过评估修复所需的细胞信号事件来更广泛地评估。研究表明，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，其作用强于被广泛研究的GAG、硫酸肝素（HS）。本研究的目的是进一步表征DS对体外成纤维细胞生长因子信号传导的影响，并将这些发现推断到人类生理环境中。总体假设是FGF-10和FGF-22倾向于DS而不是HS，并且伤口环境中DS结合蛋白的数量可以显著影响组织修复。在Aim I中，我们将使用体外生化方法确定在FGF10和FGF-22介导的信号传导中，DS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。Aim II旨在通过体内分子方法评估人体伤口液，以识别与DS比HS结合更强的FGFs。此外，还将评估DS对FGF定位和伤口闭合的影响。该实验将阐明DS在最佳组织修复所需的fgf细胞信号事件中的作用，并表征DS结合蛋白的表达模式，最终可用于病理疾病的诊断标记。临床研究表明，GAGs在皮肤病的发病机制中发挥作用。发现必需的FGFR2-IIIB配体对DS的依赖性可以预测DS合成或降解的紊乱会破坏正常的伤口修复。具体来说，我们怀疑在异常伤口中存在的DS平衡将不同于在正常修复中观察到的平衡，这得到了一些临床研究的支持。因此，分析正常或患病人体伤口液中DS的水平和/或硫酸化是本研究的一个前瞻性临床方向，这可能为伤口异常愈合提供诊断工具，并为促进伤口正常修复提供新的治疗方法。总的来说，所提出的技术允许接触新的分子和生化方法来分析皮肤伤口愈合中的蛋白质和碳水化合物功能和先天免疫。

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.