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 大规模放射学或核事件发生后，可能成千上万的人接触 电离辐射，并需要随后的剂量依赖性医疗管理。收集至关重要 并在第一周内尽快分析人类生物流体（例如血液，尿液，唾液） 准确的剂量预测和早期分类决策。需要FDA批准的体外诊断 高通量生物测量设备，能够精确地确定过去的辐射暴露 准确。在哥伦比亚大学中心高通量辐射生物测量中心 对于针对辐射（CMCR）的医疗对策，我们已经开发了快剂量（荧光灯 剂量测定的自动筛选工具）测定系统，以测量辐射响应蛋白 人类外周血样品，用于回顾性辐射剂量的回顾性估计。蛋白质面板也 包括血清细胞亚型的生物标志物，以反映血液学敏感性和损伤。快速 剂量测定系统旨在作为21 CFR 809.3定义的体外诊断装置（IVD）。这 平台使用商业成像流式细胞仪系统（Imagestream®X）和相关图像数据 探索和分析软件（Ideas®）以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 此处计划的研究旨在开发和优化我们的快剂量分析系统，以准确 估计在电离照射后，在人淋巴细胞中吸收剂量和评估造血损伤。这 第一个目标是建立我们当前的生物标志物验证数据，以通过 提前的过程。我们已经使用了人体离体模型和人性化小鼠（Hu-NSG）和非 - 人类灵长类动物（NHP）模型，以验证血清细胞中生物标志物表达和放射敏感性 急性电离辐射暴露。这里提出的具体目的旨在：优化测定 协议并确定生物标志物剂量/时间动力学，以便在体外和测试中进行准确的剂量预测1） 变异，2）持有内变异和3）实验室间变异性（AIM 1）；测试特定的效果 混杂因素：年龄和性别，与种系BRCA1/2致病变体的继承以及炎症和创伤 在辐照之前和之后的生物标志物反应上（AIM 2）；测量生物标志物水平和时间动力学 基于外周血白细胞计数和茎和茎，体内与造血损伤相关 HU-NSG小鼠的骨髓中的祖细胞水平（AIM 3）和开发数学模型（使用 机器学习和回归技术）选择最佳的快速剂量生物标志物及其组合 用于基于这些生物标志物的离体和体内剂量反应的剂量预测（AIM 4）。 我们对未来发展的愿景是开发更简化，更快的快速剂量分析系统 因此，可以开发和过渡生物标志物，以在护理点（POC）设备中使用。