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: 10611451
• 负责人: Evagelia Christos Laiakis
• 金额: $ 56.19万
• 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/10611451
• 关键词: Automation; Binding; Biological Assay; Biological Markers; Blood; Blood specimen; Clinical; Clinical Research; Communities; Databases; Dependence; Development; Devices; Dose; Dryness; 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; biomarker discovery; biomarker identification; biomarker panel; biosignature; cohort; comparison control; cost efficient; design; experience; experimental study; flexibility; human error; instrument; liquid biopsy; manufacture; 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; technology platform; 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. !

该提案建立在我们在未靶向和有针对性代谢组学方面的代谢组学专业知识上 产生高度敏感和定量的靶向多重测定。产生这样的想法 易于获得的生物流体（尿，血液，唾液）的辐射评估和辐射损伤的测定是 迅速确定个人暴露的程度，并区分担心的井和 可能需要医疗干预的人。将采取高度定量的方法 通过液相色谱串联质谱法（LC-MS/MS）来量化每个已经鉴定的 每个生物流体中的辐射生物标志物。目前，此类仪器通常在临床实验室中使用， 因此，最大化可用资源以快速评估成千上万的人 紧急情况。基于灵敏度，高信噪比的标准，矩阵的低信号抑制 效果和高倍数变化与对照组或对代谢物之间的关系相比 将组装生物签名并计算浓度。合并的生物签名将开发 在多重测定中，有效地将样品制备之间的时间减少到结果。目标是 证明这种多重测定方法有可能在紧急情况下部署 预定的临床实验室网络，可以接受并迅速处理大量的临床实验室网络 样品。虽然最终目标是在所有情况下都有这样的小组的预测性，甚至有限的错误 正率将促进在大规模伤亡情况下评估辐射损伤：例如1％假 正率将使需要进一步评估的个人数量减少100倍。另外，这个 测定将是灵活的，因为它可以用特异性和辐射质量富含生物标志物。这 应用程序还结合了我们合作者的工程经验和能力，以进一步发展 预处理设备的目的是在运输到临床设施期间稳定样品。这 要制造的材料还将旨在丰富与辐射相关的代谢物和 从少量的生物流体（尿液，血清，全血，唾液）中有效提取它们 稳定的干膜。在96井板中的此类材料组装将进一步减少样品 准备时间并最大程度地减少与样品制备相关的人为错误。我们独特的方法 将LC-MS/MS应用程序与预处理材料相结合，旨在将该技术从 技术开发的可行性阶段，满足辐射损伤快速方法的需求 评估。 呢