Targeted Biomarker Panels and Pre-processing Device for the Rapid Assessment of Radiation Injury in Easily Accessible Biofluids

用于快速评估易于获取的生物流体中的辐射损伤的靶向生物标记物组和预处理装置

• 批准号: 10459218
• 负责人: Evagelia Christos Laiakis
• 金额: $ 66.37万
• 依托单位: GEORGETOWN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-07-27 至 2026-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10459218
• 关键词: Automation; Biological Assay; Biological Markers; Blood; Blood specimen; Clinical; Clinical Research; Communities; Databases; Dependence; Development; Devices; Dose; Emergency Situation; Engineering; Environment; Evaluation; Exhibits; Exposure to; Funding; Future; Gel; Generations; Goals; Human; Individual; Injury; Intention; Intervention; Ionizing radiation; Laboratories; Liquid Chromatography; Medical; Membrane; Metabolic; Methods; Modeling; Mus; Noise; Phase; Preparation; Process; Radiation; Radiation Accidents; Radiation Injuries; Radiation exposure; Radiology Specialty; Readiness; Reproducibility; Research; Resources; Saliva; Sampling; Serum; Signal Transduction; Solid; Specificity; System; Techniques; Technology; Temperature; Testing; Textiles; Time; Tissues; Urine; Validation; Whole Blood; base; biomarker discovery; biomarker panel; biosignature; cohort; cost efficient; design; experience; experimental study; flexibility; human error; instrument; liquid biopsy; mass casualty; medical countermeasure; metabolomics; multiplex assay; new technology; nonhuman primate; novel; predictive panel; preservation; prototype; radiation-induced injury; rapid technique; response; saliva sample; stability testing; tandem mass spectrometry; targeted biomarker; technology development; urinary; virtual

This proposal builds upon our metabolomics expertise in untargeted and targeted metabolomics for the generation of highly sensitive and quantitative targeted multiplex assays. The idea behind generating such assays for radiation assessment and radiation injury in easily accessible biofluids (urine, blood, saliva) is to rapidly determine the extent of exposure of an individual and distinguish between the worried well and the exposed individuals that may require medical intervention. Highly quantitative approaches will be undertaken through liquid chromatography tandem mass spectrometry (LC-MS/MS) to quantify each already identified radiation biomarkers in each biofluid. Such instruments are currently used routinely in clinical laboratories, therefore maximizing the available resources for rapid evaluation of thousands of individuals during an emergency. Based on criteria for sensitivity, high signal-to-noise ratio, low signal suppression from matrix effects, and high fold changes compared to controls or relationships between pairs of metabolites, biosignatures will be assembled and concentrations calculated. The combined biosignature will be developed in a multiplex assay, effectively reducing the time between sample preparation to results. The goal is to demonstrate that this multiplex assay method has the potential to be deployed in the case of an emergency to a pre-determined network of clinical laboratories that can accept and rapidly process a high volume of samples. While the ultimate goal will be for such a panel to be predictive in all cases, even a limited false positive rate would facilitate assessment of radiation injury in a mass casualty scenario: e.g. a 1% false positive rate would reduce the number of individuals needing further evaluation by 100-fold. Additionally, this assay will be flexible as it could be enriched with biomarkers for specificity and radiation quality. This application also combines the engineering experience and capabilities of our collaborators to further develop pre-processing devices with the intention of stabilizing the sample during transport to a clinical facility. The materials to be fabricated will also aim to enrich the biosignature for the radiation-related metabolites and extract them effectively from small amounts of a biofluid (urine, serum, whole blood, saliva), transported as a stable dry membrane. Assembly of such materials in a 96 well plate will further decrease the sample preparation time and minimize human error associated with sample preparation. Our unique approach to combine LC-MS/MS applications with pre-processing materials will aim to move this technology from the feasibility stage to technology development, satisfying the needs for rapid methods for radiation injury assessment. !

该提案基于我们在非靶向和靶向代谢组学方面的代谢组学专业知识， 产生高灵敏度和定量的靶向多重测定。产生这样一种 用于在容易获得的生物流体（尿液、血液、唾液）中进行辐射评估和辐射损伤的测定是为了 快速确定个人的暴露程度，并区分担心的井和 可能需要医疗干预的暴露个体。将采用高度量化的方法 通过液相色谱串联质谱法（LC-MS/MS）对已鉴别的每种药物进行定量 放射性生物标记物。这种仪器目前在临床实验室中常规使用， 因此，最大限度地利用现有资源，在一个 紧急情况基于灵敏度、高信噪比、矩阵低信号抑制的标准 效应和与对照相比的高倍数变化或代谢物对之间的关系， 将收集生物特征并计算浓度。将开发组合生物特征 在多重分析中，有效地减少了样品制备到结果之间的时间。目标是 证明这种多重检测方法有可能在紧急情况下部署， 预先确定的临床实验室网络，可以接受并快速处理大量 样品虽然最终的目标将是这样一个小组在所有情况下都是预测性的，即使是有限的错误， 阳性率将有助于评估大规模伤亡情况下的辐射损伤：例如， 阳性率将使需要进一步评估的人数减少100倍。而且这个 分析将是灵活的，因为它可以富集生物标志物的特异性和辐射质量。这 应用程序还结合了我们合作者的工程经验和能力，以进一步开发 预处理装置，目的是在运输到临床设施期间稳定样品。的 所要制造的材料还将旨在丰富与辐射有关的代谢物的生物特征， 从少量的生物流体（尿液，血清，全血，唾液）中有效地提取它们，作为 稳定干膜。在96孔板中组装这些材料将进一步减少样品 缩短样品制备时间，并最大限度地减少与样品制备相关的人为错误。我们独特的方法， 联合收割机将LC-MS/MS应用与预处理材料相结合，旨在将这项技术从传统的 技术开发的可行性阶段，满足辐射损伤快速检测方法的需要 考核 !