Rapid Automated High-Throughput Radiation Biodosimetry

快速自动化高通量辐射生物剂量测定

• 批准号: 8012187
• 负责人: DAVID JONATHAN BRENNER
• 金额: $ 58.76万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-08-01 至 2015-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8012187
• 关键词: Accounting; Acute; Address; Animal Organ; Automation; Biological Assay; Biological Markers; Blood; Blood specimen; Body part; Cancer Patient; Cell division; Cessation of life; Chromosome Deletion; Chromosome abnormality; Complex; DNA Damage; DNA Repair; Data; Development; Devices; Dose; Dose-Rate; Ear; Erythema; Event; Explosion; Exposure to; Fast Neutrons; Gamma Rays; Goals; Hematopoietic; Hour; Human; In Vitro; Inbred Mouse; Inbred Strains Mice; Individual; Investigation; Kinetics; Lead; Lethal Dose 50; Link; Lymphocyte; Measurement; Measures; Micronucleus Tests; Mind; Modification; Mononuclear; Mouse Strains; Mus; Nature; Neutrons; Nuclear; Organ; Performance; Photons; Population; Positioning Attribute; Radiation; Radiation Injuries; Radiation Tolerance; Radiation therapy; Radioactive; Radioisotopes; Readiness; Relative Biological Effectiveness; Research; Residual state; Running; Sampling; Security; Series; Signal Transduction; Simulate; System; Techniques; Time; Triage; Uncertainty; Woman; Work; base; biodosimetry; breast lumpectomy; carcinogenesis; dirty bomb; high throughput screening; in vivo; interest; irradiation; malignant breast neoplasm; man; micronucleus; reconstruction; repaired; response; statistics; tool

Our overriding theme is ultra high-throughput radiation biodosimetry. It is well established that this is a central and necessary component of any effective response to a large scale radiological event. The RABIT (Rapid Automated Biodosimetry Tool) system developed to date in Project 1 has a current throughput of 6,000 samples / day, and a projected (2010) throughput of 30,000 samples / day. These throughputs were achieved by complete robotically-based automation of standard micronucleus and Y-H2AX assays. One first main renewal theme, "Beyond Simple Exposures", is to assess the significance of various realistic radiation event scenarios, in particular protraction of radiation exposure, partial-body exposure, internal emitters, and neutron exposure. Our goal is to understand the responses of the standard RABIT assays to these types of exposures, and quantify the uncertainties associated with dose reconstruction in these scenarios. The assays will be done both with irradiated mice and with ex-vivo irradiated human blood, with the unique irradiation facilities of the Irradiation Core, allowing sample sharing with the other two Projects. While the micronucleus assay has high throughput and excellent post-irradiation stability, its assay time is ~3 days, due to the need to stimulate cell division in vitro. We have preliminary evidence of the practicality of a micronucleus assay using /T?ononi;c/ear lymphocytes, which have already divided in vivo. Our second theme is to develop and assess this system which, with full automation, would reduce the assay time to ~3 hrs Having developed high-throughput systems using various biomarkers for biodosimetry, we are in a unique position to probe the application of these high-throughput biomarkers for predicting inter-individual sensitivity to acute radiation injury. Thus our final renewal theme is "Beyond Dose: Towards Individual Radiosensitivity". The six high-throughput automated RABIT endpoints that we will use are baseline micronuclei, ex-vivo radiation-induced micronuclei, ex-vivo radiation-induced y-H2AX, ex-vivo radiation-induced chromosomal deletions, DNA damage repair kinetics (with Y-H2AX, ATM, 53BP1, and Mdcl foci), and residual damage after 24 h. The practical use of this number of assays is made possible by their being fully automated within the RABIT. Studies will be done with inbred mice with differing acute radiosensitivities, and by assessing associations between these endpoints and acute erythema in 500 irradiated breast cancer patients.

我们最重要的主题是超高通量辐射生物剂量测定。众所周知，这是一个 对大规模放射性事件作出任何有效反应的核心和必要组成部分。兔子 项目 1 迄今为止开发的（快速自动生物剂量测定工具）系统的当前吞吐量为 6,000 个样本/天，预计（2010 年）吞吐量为 30,000 个样本/天。这些吞吐量是 通过基于机器人的标准微核和 Y-H2AX 检测的完全自动化来实现。 第一个主要更新主题“超越简单的曝光”是评估各种现实的意义 辐射事件情景，特别是长期辐射暴露、局部身体暴露、内部辐射 发射器和中子暴露。我们的目标是了解标准 RABIT 检测的反应 这些类型的暴露，并量化与这些剂量重建相关的不确定性 场景。这些测定将使用受辐射的小鼠和离体受辐射的人血液进行， 辐照核心独特的辐照设施，允许与其他两个项目共享样品。 虽然微核测定具有高通量和出色的辐照后稳定性，但其测定时间很长 ~3天，由于需要刺激体外细胞分裂。我们有初步的实用性证据 使用已在体内分裂的/T?ononi;c/ear 淋巴细胞进行微核测定。我们的第二个 主题是开发和评估该系统，该系统完全自动化，可将检测时间缩短至约 3 小时 我们开发了使用各种生物标志物进行生物剂量测定的高通量系统，我们处于独特的地位 探索这些高通量生物标志物在预测个体间敏感性中的应用 致急性放射损伤。因此，我们最终的更新主题是“超越剂量：迈向个人放射敏感性”。 我们将使用的六个高通量自动化 RABIT 端点是基线微核、体外 辐射诱导的微核、离体辐射诱导的 y-H2AX、离体辐射诱导的染色体 缺失、DNA 损伤修复动力学（使用 Y-H2AX、ATM、53BP1 和 Mdcl 焦点）和残余损伤 24小时后。如此数量的测定的实际使用是通过它们在内部完全自动化而成为可能的。 兔子。研究将用具有不同急性放射敏感性的近交小鼠进行，并通过评估 这些终点与 500 名接受辐射的乳腺癌患者的急性红斑之间的关联。