Developing biomarkers of healing and non-healing VLUs

项目摘要 该项目建议使用从废弃的伤口敷料中捕获的伤口液体分析来开发 可以预测愈合结果和监测静脉性小腿溃疡(VLU)愈合进程的生物标志物。 VLU代表发生在慢性静脉环境中的结扎区的慢性溃疡的特定病例 占所有慢性腿部溃疡的70%左右。考虑到人口老龄化是 令人担忧的是：更多的溃疡出现在病情较重的患者身上，而这些患者的治疗费用越来越高。 尽管最近取得了进展，但超过50%的VLU患者未能在标准护理下痊愈。其中一个主要的 改善结果的障碍是无法及早预测谁会和谁不会对标准做出反应 关爱。因此，有一个迫切的和未得到满足的需求来预测愈合结果和区分愈合。 来自不能治愈的VLU。由于伤口内分子的动态变化，其微环境可以 测试和监测，这为获得有关伤口状态的关键信息提供了机会 生物学。此外，量化这种反映伤口愈合状况的分子变化将提供 能够指导治疗方法的重要工具，直接影响临床结果。因此，两者都具有预测性 需要监测生物标志物才能有效地实现预期的治疗定制：1)最大限度地提高 治疗结果，2)只针对需要治疗的患者进行更积极的治疗，3)发展 针对每个VLU的个性化方法。在最初的发现阶段，我们将招募30名VLU患者(10名治疗师 和20名非治疗者)，以确定一组预测愈合结果的蛋白质生物标记物(目标1)；那些可以 作为监测生物标志物(目标2)。我们将从废弃的伤口敷料中提取伤口液，并对 通过使用LC-MS/MS(LC-MS/MS)分析蛋白质并将它们与第4周的愈合结果相关联。我们有 生成了初步数据，支持整体方法的可行性，并确认伤口敷料 任何类型的(实时分析或冷冻储存)都可用于生物标记物的定量。我们有 还鉴定了在伤口敷料的中心相对于周边的分子在空间上的不同定位， 从而产生分别反映伤口床和伤口边缘的生物的特定轮廓。这个项目 将提供一个独特的机会，利用容易获得的废弃伤口敷料作为 可用于生物标志物检测的伤口生物材料(渗出物)。它不仅将识别和验证特定的 一组生物标记物，可以预测愈合结果和/或监测愈合进程，但也将提供 易于实施的简化检测方法。

Project Summary This project proposes to use wound fluid analyses captured from discarded wound dressings to develop biomarkers that can predict healing outcome and monitor healing progression of Venous Leg Ulcers (VLUs). VLUs represent a specific case of chronic ulcers occurring in the gaiter area in the setting of chronic venous insufficiency and account for about 70% of all chronic leg ulcers. Considering the aging of the population a concerning picture emerges: more ulcers occurring in sicker patients which are ever more expensive to treat. Despite recent advances, more than 50% of patients with VLUs fail to heal with standard care. One of the major obstacles to improving outcomes is the inability to predict early on who will and who will not respond to standard of care. Therefore, there is an urgent and unmet need to predict healing outcomes and to differentiate healing from non-healing VLUs. Due to dynamic molecular changes within the wound, its microenvironment can be tested and monitored, which offers an opportunity to obtain key information regarding the status of wound biology. Moreover, quantification of such molecular changes that reflect wound healing status would provide important tools that can guide treatment approach, directly impacting the clinical outcomes. Thus, both predictive and monitoring biomarkers are needed to enable prospective tailoring of therapies to efficiently: 1) maximize the treatment outcomes, 2) target more aggressive treatment to only those patients who need it and 3) develop personalized approaches to each VLU. In the initial discovery phase, we will enroll 30 VLU patients (10 healers and 20 non-healers) to identify a set of protein biomarkers that predict healing outcomes (Aim 1); those that can serve as monitoring biomarkers (Aim 2). We will extract wound fluid from discarded wound dressings and quantify proteins by using mass spectrometry (LC-MS/MS) and correlate them to healing outcomes at week 4. We have generated preliminary data that support feasibility of the overall approach and confirm that wound dressings (either analyzed in real time or stored frozen) of any type can be utilized for biomarker quantification. We have also identified spatially distinct localization of molecules at the center vs perimeter of the wound dressing, resulting in specific profiles that reflect the biology of the wound bed and wound edge respectively. This project will provide a unique opportunity to capitalize on easy access of discarded wound dressings as a source of wound biomaterial (exudate) that can serve for biomarker detection. It will not only identify and validate specific set of biomarkers that can predict healing outcome and/or monitor healing progression but will also provide streamlined method of detection that can be readily implemented.