Inflammation-related gene biomarkers in human diabetic foot ulcer healing

人类糖尿病足溃疡愈合中的炎症相关基因生物标志物

• 批准号: 10658986
• 负责人: Kara Lorraine Spiller
• 金额: $ 46.45万
• 依托单位: DREXEL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-07-04 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10658986
• 关键词: Area; Base Ratios; Biological Markers; Cells; Chronic; Clinical; Clinical Trials; Complex; Debridement; Detection; Diabetic Foot; Diabetic Foot Ulcer; Ensure; Foundations; Gene Expression; Gene Expression Profiling; Genes; Goals; Human; Impaired wound healing; Impairment; In Vitro; Individual; Inflammation; Inflammatory; Link; Lower Extremity; Measurement; Measures; Methods; Patients; Phase; Phase II Clinical Trials; Process; Prognostic Marker; Quality Control; Quantitative Reverse Transcriptase PCR; RNA; ROC Curve; Reproducibility; Research; Resolution; Sampling; Sensitivity and Specificity; Shipping; Specificity; Standardization; Techniques; Time; Tissue Banks; Tissue Model; Tissue Sample; Ulcer; United States; Visit; Work; clinical effect; cohort; collagen scaffold; cost; diabetic patient; diabetic wound healing; healing; immune activation; indexing; limb amputation; meetings; mortality; non-healing wounds; patient variability; personalized medicine; predict responsiveness; predictive marker; prognostic; prognostic value; responders and non-responders; response; sample collection; secondary analysis; standard of care; wound; wound care; wound healing

Project summary Diabetic foot ulcers (DFUs) occur in 15% of diabetic patients, leading to over 82,000 lower limb amputations annually in the United States and a 5-year mortality rate of up to 74%. The reasons for impaired DFU healing are complex, but the downstream effects of chronic inflammation are major contributors. Our research has shown that while initial pro-inflammatory activation of immune cells is critical for the initiation of healing processes, prolonged activation directly impairs wound healing. Recognizing that transition from the early inflammatory phase to the late resolution phase is required for successful healing, we developed a composite biomarker that uses the change in the ratio of 4 early stage pro-inflammatory gene markers to 3 late stage inflammation-resolution gene markers over 4 weeks to predict responsiveness to treatment. By using a ratio of these early-stage to late-stage genes, referred to here as the Inflammation Index, higher values indicate wounds that are earlier in the healing process (and further from healing), while lower values indicate a wound that is later in the healing process (and closer to healing). Thus, a decrease in this index over time is linked to healing, while an increase is linked to exacerbating inflammation and non-healing. In our preliminary studies (3 cohorts of 21 subjects), the change in the Inflammation Index correctly predicted healing in 10 out of 10 healing wounds (responders), and it correctly predicted non-healing in 9 out of 11 non-healing wounds (non-responders), with an area under the ROC curve of 0.9. This biomarker utilizes debrided wound tissue so that it can be easily incorporated into standard wound care practice without adding any new techniques or time into the visit. Expression of the 7 genes that comprise the composite biomarker is measured using qRTPCR, a widely available, low-cost, and reliable technique. Finally, the use of a ratio self-normalizes the gene expression values to reduce patient-to-patient variability and increase reproducibility. In this project, we will develop and internally validate detection of this composite ratio-based biomarker; establish proof of concept of its prognostic utility; lay the foundation for a specific Context of Use (COU); and ultimately ensure that is ready for Phase II clinical trials. The proposed COU is a prognostic biomarker to identify individuals who are not likely to heal their ulcer when treated with the standard of care, for use in the personalization of treatment and/or the refinement of entry criteria for clinical trials of new treatments. In the R61 phase of this project, we will optimize biomarker measurement and standardization, and determine the quality control (QC) metrics that can be used to determine if the biomarker is accurately measured. We will also validate storage/shipping conditions and measure reliability and reproducibility. After meeting rigorous milestones the biomarker will progress to the R33 phase of the project, in which we will measure its ability to predict healing in response to the standard of care, in order to ultimately personalize treatment for patients with hard-to-heal ulcers and to refine entry criteria for clinical trials of new treatments.

项目摘要 糖尿病足溃疡（DFU）发生在15%的糖尿病患者中，导致超过82，000例下肢溃疡。 在美国，每年有100例截肢，5年死亡率高达74%。受损的原因 DFU愈合是复杂的，但慢性炎症的下游效应是主要贡献者。我们 研究表明，虽然免疫细胞的初始促炎性激活对于启动炎症反应是关键的， 在愈合过程中，延长的活化直接损害伤口愈合。认识到， 早期炎症阶段到晚期消退阶段是成功愈合所必需的，我们开发了一种 使用4种早期促炎基因标志物与3种晚期促炎基因标志物的比率的变化的复合生物标志物 在4周内分期炎症消退基因标记物，以预测对治疗的反应性。通过使用 这些早期与晚期基因的比率，在此称为炎症指数，较高的值表明 在愈合过程中较早（并且离愈合较远）的伤口，而较低的值表示伤口 这是在愈合过程中的后期（更接近愈合）。因此，这一指数随时间的下降与以下因素有关： 愈合，而增加与炎症加剧和不愈合有关。在我们的初步研究（3 21名受试者的队列），炎症指数的变化正确预测了10/10的愈合 伤口（响应者），并且它正确地预测了11个未愈合伤口（无响应者）中的9个未愈合， ROC曲线下面积为0.9。该生物标志物利用清创的伤口组织，使得其可以容易地被清除。 将其纳入标准伤口护理实践中，而无需增加任何新技术或时间。 使用qRTPCR测量包含复合生物标志物的7个基因的表达， 成本低，技术可靠。最后，比率的使用自归一化基因表达值以减少 患者与患者之间的变异性并提高可重复性。 在这个项目中，我们将开发和内部验证这种基于复合比率的生物标志物的检测; 建立其预后效用的概念证明;为特定的使用环境（COU）奠定基础;以及 最终确保为II期临床试验做好准备。拟议的COU是一种预后生物标志物， 当用标准护理治疗时，不太可能治愈其溃疡的个体， 个性化治疗和/或改进新治疗临床试验的进入标准。 在该项目的R61阶段，我们将优化生物标志物测量和标准化， 确定可用于确定生物标志物是否被准确测量的质量控制（QC）指标。 我们还将验证储存/运输条件，并测量可靠性和再现性。会晤后 严格的里程碑生物标志物将进展到项目的R33阶段，在此阶段，我们将测量其 能够根据护理标准预测愈合，以便最终个性化治疗 难以治愈的溃疡患者，并完善新治疗方法临床试验的入选标准。