Development of FAST-DOSE assay system for the rapid assessment of acute radiation exposure, individual radiosensitivity and injury in victims for a large-scale radiological incident

开发快速剂量测定系统，用于快速评估大规模放射事件受害者的急性辐射暴露、个体放射敏感性和损伤

• 批准号: 10558479
• 负责人: Helen C Turner
• 金额: $ 58.47万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-02-01 至 2025-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10558479
• 关键词: Academic Medical Centers; Acute; Age; Algorithms; Benchmarking; Biological Assay; Biological Markers; Blood; Blood Proteins; Blood specimen; Bone Marrow; Bone Marrow Stem Cell; Burn injury; Cell Culture Techniques; Cellular Structures; Color; Computer software; Confidence Intervals; Data; Data Analyses; Detection; Development; Devices; Diagnostic Equipment; Dose; Exposure to; FDA approved; Flow Cytometry; Fluorescence; Future; Hematology; Hematopoietic; Human; Image; Imagery; Immune; Immunoassay; In Vitro; Individual; Industrial Accidents; Inflammation; Inherited; Injury; Ionizing radiation; Kinetics; Laboratories; Leukocytes; Linear Regressions; Lymphocyte; Machine Learning; Mass Screening; Measures; Medical; Medical center; Modeling; Mus; Noise; Nuclear Accidents; Pathogenicity; Patients; Performance; Persons; Physiological; Population; Process; Proteins; Protocols documentation; Radiation; Radiation Accidents; Radiation Dose Unit; Radiation Tolerance; Radiation exposure; Reproducibility; Research Design; Roentgen Rays; Saliva; Screening procedure; Surface Antigens; System; Techniques; Testing; Time; Toxic effect; Trauma; Triage; Uncertainty; Urine; Variant; Vision; White Blood Cell Count procedure; absorption; biodosimetry; biomarker identification; biomarker panel; biomarker performance; biomarker validation; brca gene; data exploration; design; dirty bomb; dosimetry; humanized mouse; in vitro testing; in vivo; in-vitro diagnostics; ionization; irradiation; machine learning algorithm; mathematical model; medical countermeasure; micronucleus; nonhuman primate; nonlinear regression; performance tests; peripheral blood; point of care; predictive modeling; protein biomarkers; response; response biomarker; sex; stem; stem cells

Summary Following a large scale radiological or nuclear event, hundreds of thousands of people may be exposed to ionizing radiation/s and require subsequent dose-dependent medical management. It will be crucial to collect and analyze human biofluids (such as blood, urine, saliva) as soon as possible within the first week for accurate dose prediction and early triage decision. There is a need for FDA-approved in vitro diagnostic high-throughput biodosimetry devices with the ability to determine past radiation exposure with precision and accuracy. At the Center for High Throughput Radiation Biodosimetry, the Columbia University Center for Medical Countermeasures against Radiation (CMCR), we have developed FAST-DOSE (Fluorescent Automated Screening Tool for Dosimetry) assay system, to measure radiation-responsive proteins in human peripheral blood samples for retrospective estimation of radiation dose. The protein panel also includes biomarkers for blood leukocyte subtypes to reflect hematological sensitivity and injury. The FAST- DOSE assay system is intended as an in vitro diagnostic device (IVD) as defined by 21 CFR 809.3. The platform uses a commercial imaging flow cytometry system (ImageStream®X) and associated Image Data Exploration and Analysis Software (IDEAS®) to rapidly quantify changes in biomarker expression levels within specific cellular structures using fluorescent imagery and algorithms for estimation of absorbed dose. The studies planned here are designed to develop and optimize our FAST-DOSE assay system to accurately estimate absorbed dose and assess hematopoietic injury in human lymphocytes after ionizing irradiation. The first objective is to build on our current biomarker validation data for early engagement with the FDA via the pre-submission process. We have used the human ex vivo model and humanized mouse (Hu-NSG) and non- human primate (NHP) models to validate biomarker expression and radiosensitivity in blood leukocytes after acute ionizing radiation exposure. The Specific Aims proposed here are designed to: optimize the assay protocol and identify biomarker dose/time kinetics for accurate dose predictions in vitro and test 1) inter-donor variation, 2) intra-donor variation and 3) inter-laboratory variability (Aim 1); test the effect of specific confounders: age and sex, inheritance with germline BRCA1/2 pathogenic variant, and inflammation and trauma on the biomarker response, before and after irradiation (Aim 2); measure biomarker levels and time kinetics in vivo and correlate with hematopoietic injury, based on peripheral blood leukocyte counts, and stem and progenitor cell levels in the bone marrow of Hu-NSG mice (Aim 3) and, develop mathematical models (using machine learning and regression techniques) to select the best FAST-DOSE biomarkers and their combinations for generating dose predictions based on the ex vivo and in vivo dose response of these biomarkers (Aim 4). Our vision for future development is to develop a more simplified, faster rapid FAST-DOSE assay system whereby the biomarkers could be developed and transitioned for use in a point-of-care (POC) device.

概括 大规模放射性或核事件发生后，数十万人可能会受到辐射 电离辐射，并需要随后的剂量依赖性医疗管理。收集起来将至关重要 并在第一周内尽快分析人体生物体液（如血液、尿液、唾液） 准确的剂量预测和早期分诊决策。需要 FDA 批准的体外诊断 高通量生物剂量测定设备，能够精确确定过去的辐射暴露 和准确性。哥伦比亚大学高通量辐射生物剂量测定中心 针对放射线医疗对策（CMCR），我们开发了FAST-DOSE（荧光 用于剂量测定的自动筛选工具）测定系统，用于测量辐射响应蛋白 人体外周血样本用于回顾性估计辐射剂量。蛋白质面板还 包括血液白细胞亚型的生物标志物，以反映血液学敏感性和损伤。快速- DOSE 测定系统旨在用作 21 CFR 809.3 定义的体外诊断设备 (IVD)。这 平台使用商业成像流式细胞术系统 (ImageStream®X) 和相关图像数据 探索和分析软件 (IDEAS®) 可快速量化生物标志物表达水平的变化 使用荧光图像和算法来估计吸收剂量的特定细胞结构。这 这里计划的研究旨在开发和优化我们的快速剂量测定系统，以准确地 估计吸收剂量并评估电离辐射后人类淋巴细胞的造血损伤。这 第一个目标是建立在我们当前的生物标志物验证数据的基础上，以便通过 预提交过程。我们使用了人类离体模型和人源化小鼠（Hu-NSG）和非 人类灵长类动物（NHP）模型，用于验证血液白细胞的生物标志物表达和放射敏感性 急性电离辐射暴露。这里提出的具体目标旨在： 优化检测 方案并确定生物标志物剂量/时间动力学，以进行准确的体外剂量预测和测试 1) 供体间 变异，2) 捐赠者内部变异和 3) 实验室间变异（目标 1）；测试具体效果 混杂因素：年龄和性别、种系 BRCA1/2 致病性变异遗传、炎症和创伤 照射前后的生物标志物反应（目标 2）；测量生物标志物水平和时间动力学 体内并与造血损伤相关，基于外周血白细胞计数以及干细胞和 Hu-NSG 小鼠骨髓中的祖细胞水平（目标 3），并开发数学模型（使用 机器学习和回归技术）来选择最佳的快速剂量生物标志物及其组合 用于根据这些生物标志物的离体和体内剂量反应生成剂量预测（目标 4）。 我们未来发展的愿景是开发更简化、更快速的 FAST-DOSE 检测系统 由此可以开发生物标记物并将其转换用于即时护理 (POC) 设备。