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.

项目概要 15% 的糖尿病患者出现糖尿病足溃疡 (DFU)，导致超过 82,000 名下肢溃疡 美国每年都会发生截肢手术，5 年死亡率高达 74%。受损原因 DFU 愈合很复杂，但慢性炎症的下游影响是主要因素。我们的 研究表明，虽然免疫细胞最初的促炎激活对于启动 愈合过程中，长时间激活会直接损害伤口愈合。认识到从 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 复合生物标志物，利用 4 个早期促炎基因标志物与 3 个晚期促炎基因标志物的比率变化 在 4 周内对炎症消退基因标记进行分级，以预测对治疗的反应。通过使用 这些早期与晚期基因的比率，此处称为炎症指数，较高的值表示 伤口处于愈合过程的早期（并且距离愈合较远），而较低的值表示伤口 那是在愈合过程的后期（并且更接近愈合）。因此，该指数随着时间的推移而下降与 愈合，而增加则与炎症加剧和不愈合有关。在我们的初步研究中（3 21 名受试者的队列），炎症指数的变化正确预测了 10 例愈合中的 10 例的愈合 伤口（有反应者），并且它正确预测了 11 个不愈合伤口（无反应者）中的 9 个不愈合， ROC 曲线下面积为 0.9。该生物标记物利用清创的伤口组织，因此可以很容易地 纳入标准伤口护理实践，无需增加任何新技术或就诊时间。 使用 qRTPCR（一种广泛使用的、 成本低，技术可靠。最后，使用比率自我标准化基因表达值以减少 患者之间的变异性并提高可重复性。 在这个项目中，我们将开发并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；和 最终确保为二期临床试验做好准备。拟议的 COU 是一种预后生物标志物，用于识别 当接受标准护理治疗时不太可能治愈溃疡的个体，用于 治疗的个性化和/或新治疗临床试验准入标准的细化。 在该项目的R61阶段，我们将优化生物标志物测量和标准化，以及 确定可用于确定生物标志物测量是否准确的质量控制 (QC) 指标。 我们还将验证存储/运输条件并测量可靠性和再现性。见面后 该生物标记物将进入该项目的 R33 阶段，在此阶段我们将测量其严格的里程碑 根据护理标准预测愈合的能力，以便最终实现个性化治疗 难以治愈的溃疡患者，并完善新疗法临床试验的准入标准。