MicroRNA-Based Radiation Biodosimeter (MiRAD)

描述(申请人提供)：该项目广泛的长期目标是开发一种光学辐射生物剂量计，它能快速(~15分钟)测量血清微小RNA(MiRNA)生物标记物的变化，以预测急性辐射综合征(ARS)。MicroRNAs(MiRNAs)是一种短的(18-22碱基)非编码RNA链，通过调节蛋白质的表达来控制一系列生物过程。暴露在辐射中可以改变与ARS相关的miRNA面板的表达，从而确立血清可以作为ARS的预测分析。为了测量这些miRNA辐射生物标志物的变化，我们将开发一种基于microRNA的辐射生物剂量计(Mirad)系统，该系统将基于磁珠的miRNA靶标自动筛选与ChromoLogic专利技术放大反射干涉分析(ARIA)相结合，用于高灵敏度和特异性地并行定量100个miRNA辐射生物标志物。在初步研究中，我们展示了该设备的新概念及其准确测量卵巢癌血清样本中毫微摩尔以下的miRNA浓度的能力，比目前的RNA定量平台(如RT-PCR)少100倍。我们使用啮齿动物血浆进行的初步研究也发现了一组对全身照射(WBI)具有剂量和时间依赖反应的miRNAs，其剂量范围与发生放射事件的医学分诊相关。根据拟议的概念验证研究，我们试图验证的假设是(1)通过分析血浆在小鼠模型中确定的miRNA小组可用于临床环境中的WBI剂量测量；(2)类似的miRNA小组可用于测量非人类灵长类动物和人类的WBI剂量；(3)最终确定的独特且具有剂量响应的miRNA标记组合将导致快速、POC血基微芯片检测结果，用于预测暴露后第一周内急性呼吸道综合征的发生，从而指导暴露对象的分配以进行及时的医疗干预。

DESCRIPTION (provided by applicant): The broad long-term goal of this project is to develop an optically based radiation biodosimeter that rapidly (~ 15 minutes) measures changes in serological microRNA (miRNA) biomarkers to predict acute radiation syndrome (ARS). MicroRNAs (miRNAs) are short (18-22 base) non- coding RNA strands that control a host of biological processes through regulation of protein expression. Exposure to radiation has been shown to alter expression of a miRNA panels linked with ARS, thereby establishing that serum could be used as a predictive assay for ARS. To measure changes in these miRNA radiation biomarkers, we will develop a MicroRNA-based Radiation Biodosimeter (MiRAD) system, which combines automated magnetic bead based screening for miRNA targets with Amplified Reflectometric Interference Analysis (ARIA), a patented ChromoLogic technology, for highly sensitive and specific quantification of 100s of miRNA radiation biomarkers in parallel. In preliminary studies, we have demonstrated the novel concept of the device and its ability to accurately measure subfemtomolar concentrations of miRNA in ovarian cancer serum samples, 100x less than current RNA quantification platforms such as RT-PCR. Our preliminary studies using rodent plasma also have identified panel of miRNAs with dose and time dependent response to whole body irradiation (WBI), in a dose range relevant to medical triage in case of a radiological event. The hypotheses we seek to validate under the proposed proof-of-concept study are (1) the miRNA panel identified in a murine model through analysis of plasma can be used to measure WBI dose in a clinical setting; (2) a similar miRNA panel can be used to measure WBI dose in nonhuman primates and humans; (3) the resulting identification of a unique and dose-responsive combination of miRNA markers will result in a rapid, POC blood-based microchip assay for anticipating ARS within the first week of exposure, thereby guiding the allocation of exposed subjects for timely medical intervention.