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; 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个下肢 每年在美国的截肢和5年的死亡率高达74％。受损的原因 DFU愈合很复杂，但是慢性炎症的下游效应是主要因素。我们的 研究表明，虽然免疫细胞的初始促炎激活对于启动至关重要 愈合过程，长时间的激活直接损害伤口愈合。认识到从 成功愈合需要早期炎症阶段到晚期分辨率阶段，我们开发了一个 复合生物标志物使用4个早期促炎基因标记的比率变化与3个晚期 阶段炎症分辨率基因标记在4周内以预测对治疗的反应。通过使用 这些早期阶段与晚期基因的比率，在此称为炎症指数，较高的值表明 在愈合过程中（以及更远的愈合）中较早的伤口，而较低的值表示伤口 这是在康复过程中（更接近康复）的后期。因此，随着时间的推移，该指数的减少与 愈合，而增加却与加剧的炎症和非污染有关。在我们的初步研究中（3 21受试者的队列），炎症指数的变化正确地预测了10分的愈合中的10个 伤口（响应者），并在11个非愈合伤口（非响应者）中正确预测了非愈合， ROC曲线下方的面积为0.9。该生物标志物利用了清创的伤口组织，因此很容易 纳入标准伤口护理实践中，而无需在访问中添加任何新技术或时间。 使用QRTPCR测量组成复合生物标志物的7个基因的表达，这是一个广泛可用的， 低成本和可靠的技术。最后，使用比率自称为基因表达值降低 患者到患者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II期临床试验做好准备。提出的cou是一种预后的生物标志物，可以识别 接受护理标准治疗时不太可能治愈溃疡的人 个性化治疗和/或新治疗临床试验的入学标准的完善。 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是否准确测量的质量控制（QC）指标。 我们还将验证存储/运输条件并衡量可靠性和可重复性。见面后 严格的里程碑生物标志物将发展到项目的R33阶段，我们将在其中衡量其 能够响应护理标准来预测治愈的能力，以最终个性化治疗 难以愈合的溃疡患者，并完善新疗法临床试验的进入标准。