Rapid Automated High-Throughput Radiation Biodosimetry

我们的首要主题是超高通量辐射生物剂量学。众所周知，这是一个 这是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 （快速自动生物剂量测定工具）系统开发的项目1中，目前的吞吐量为 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 通过标准微核和Y-H2 AX测定的完全基于机器人的自动化实现。 第一个主要的更新主题，“超越简单的暴露”，是评估各种现实的意义， 辐射事件情景，特别是辐射照射、部分身体照射、内部 发射器和中子暴露。我们的目标是了解标准RABIT检测对以下因素的反应： 这些类型的照射，并量化与这些照射中剂量重建相关的不确定性 场景将使用经辐照的小鼠和离体经辐照的人血液进行测定， 辐照堆芯独特的辐照设施，可与其他两个项目共用样品。 虽然微核试验具有高通量和极好的辐照后稳定性，但其试验时间为 ~3天，由于需要在体外刺激细胞分裂。我们有初步证据证明 微核试验使用/T？Ononi;c/耳淋巴细胞，其已经在体内分裂。我们的第二 主题是开发和评估该系统，该系统采用全自动化，可将测定时间缩短至约3小时 我们已经开发出使用各种生物标记物进行生物剂量测定的高通量系统， 位置，以探索这些高通量生物标志物的应用，用于预测个体间的敏感性 急性辐射损伤因此，我们最后的更新主题是“超越剂量：走向个体放射敏感性”。 我们将使用的六个高通量自动RABIT终点是基线微核，离体 辐射诱导微核、离体辐射诱导γ-H2 AX、离体辐射诱导染色体 缺失、DNA损伤修复动力学（使用Y-H2 AX、ATM、53 BP 1和Mdcl灶）和残留损伤 24小时后，这一数量的测定的实际使用通过它们在内部完全自动化而成为可能。 兔子研究将使用具有不同急性放射敏感性的近交系小鼠进行，并通过评估 这些终点与500名接受过放射治疗的乳腺癌患者的急性红斑之间的相关性。

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.