The roles of TfRs and HSPA5 in Pu uptake

摘要：钚（Pu）是世界上大量生产的所有锕系元素中毒性最大的元素之一。许多钚放射性同位素的半衰期非常长，它们的长期储存导致最终污染的风险，可能影响清理工人和公众。虽然过去对Pu和哺乳动物细胞之间的相互作用进行了许多研究，但这些相互作用的细胞和分子生物学方面仍未被探索。此外，研究这种相互作用的技术在过去十年中也发生了变化。我们的团队利用阿贡国家实验室（ANL）锕系元素实验室和同步加速器提供的独特资源，利用x射线荧光光谱和显微镜进行了新型实验，这些实验只能在第三代同步加速器（如ANL的先进光子源（APS））上完成。我们现在建议在2D和3D细胞培养中询问Pu与肝细胞的相互作用，探测我们根据初步和已发表的数据预测的几种Pu蛋白相互作用。为了这项工作，我们将再次使用APS的设备，但是，除了我们在出版物中使用的设备（包括该项目中大部分初步数据）之外，我们还将使用APS新建的x射线显微镜仪器，生物纳米探测器。这种新型定制仪器可用于获取低温制备的含Pu样品的层析x射线荧光数据，以验证Pu蛋白相互作用，定量和三维绘制Pu，确定其氧化态并探测其化学环境。这项工作将通过基因表达谱和蛋白质谱对暴露于Pu的细胞分泌组进行补充，以确定不同生理条件下暴露于Pu的生物标志物。

DESCRIPTION (provided by applicant): Dissecting the roles of TfRs and HSPA5 in Pu accumulation in liver Abstract: Plutonium (Pu) is one of the most toxic of all actinide elements produced in high quantities around the world. Many Pu radioisotopes have very long half-lives and their extended storage leads to risks of eventual contamination that can affect both the clean-up workers and the public. While many studies of interactions between Pu and mammalian cells were done in the past, the cell and molecular biology aspects of these interactions remain unexplored. In addition, technologies for studying such interactions have changed during the past decade. Our team has used unique resources offered by the actinide laboratory and synchrotron at Argonne National Laboratory (ANL) to carry out novel types of experiments using X-ray fluorescence spectroscopy and microscopy that can be done only at third generation synchrotrons such as the Advanced Photon Source (APS) at ANL. We now propose to interrogate Pu interaction with liver cells in 2D and 3D cell cultures, probing several Pu-protein interactions we predict based on our preliminary and published data. For this work we will once again use the facilities at the APS but, in addition to equipment we used in publications that comprise the bulk of the preliminary data in this project, we will also use a newly built X-ray microscopy instrument, the Bionanoprobe also at the APS. This new, custom built instrument can be used to obtain tomographic X-ray fluorescence data on cryogenically prepared Pu-containing specimens in order to validate Pu-protein interactions, quantitate and map Pu in 3D, determine its oxidation state and probe its chemical environment. This work will be complemented by gene expression profiling and protein mass spectrometry of the Pu exposed cells' secretome in the interest of identifying biomarkers of Pu exposure under different physiological conditions.