Rapid Automated High-Throughput Radiation Biodosimetry

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，这是一项 这是对大规模辐射事件作出有效反应的核心和必要组成部分。RABIT (快速自动化生物剂量测定工具)到目前为止在项目1中开发的系统的吞吐量为 每天6,000个样品，预计(2010年)日吞吐量为30,000个样品。这些吞吐量是 通过标准微核和Y-H_2AX分析的完全机器人自动化实现。 第一个主要的更新主题是“超越简单的暴露”，是评估各种现实的意义。 辐射事件情景，特别是辐射暴露的延长、部分身体暴露、内部 发射器和中子暴露。我们的目标是了解标准RABIT分析对 这些类型的照射，并量化与这些类型的剂量重建相关的不确定性 场景。这些检测将在辐照的小鼠和体外辐照的人血中进行， 辐照核心独特的辐照设施，允许与其他两个项目共享样品。 虽然微核试验具有高通量和良好的辐照后稳定性，但其测定时间为 ~3天，由于需要刺激细胞在体外分裂。我们有初步证据证明…… 用已经在体内分裂的/T？ononi；c/EAR淋巴细胞进行微核试验。我们的第二个 主题是开发和评估这一系统，该系统在完全自动化的情况下，将测试时间减少到~3小时 在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后，我们处于一个独特的 探索这些高通量生物标志物在预测个体间敏感性方面的应用 到急性辐射损伤。因此，我们最后的更新主题是“超越剂量：迈向个人的辐射敏感性”。 我们将使用的六个高通量自动RABIT端点是基线微核、体外 辐射诱导微核、体外辐射诱导的γ-H_2AX、体外辐射诱导的染色体 缺失、DNA损伤修复动力学(与Y-H_2AX、ATM、53BP1和MdCL焦点)，以及残留损伤 24小时后。这一数量的分析的实际使用是因为它们在 RABIT。研究将用具有不同急性辐射敏感性的近交系小鼠进行，并通过评估 500名接受放射治疗的乳腺癌患者中，这些终点与急性红斑的关系。

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.