Correlative Nano-Characterisation for Prostate Cancer

假定前列腺癌进展为转移状态是通过外泌体，由此外泌体起作用以调节身体的组织以使它们准备好接受和支持癌细胞生长。然而，解决这一假设已被证明具有挑战性，因为存在许多形式的相似大小（<150 nm）的细胞外囊泡，并且当在收集包括外来体的异源群体中实验性地分离细胞外囊泡时。因此，在实验之前确定富集的外泌体级分是必要的，这需要结构分析（通过cryoTEM）以及组成分析。通过我们最近的初步研究，我们现在可以通过cryoTEM将荧光标记的外泌体与结构扫描光学相关。在提供重要的结构信息的同时，我们希望扩展这项技术，以在单个外泌体水平上提供完整的成分分析，这是该项目的首要目标。第二个目标是确定外泌体进入组织的入口。这需要目前缺乏的机械和生物物理理解。该项目将围绕UoN新的低温制备设施测试不同的新工作流程方法，包括大量未经尝试的相关工作流程，以实现科学目标。该项目得到了微米和纳米研究中心最新的150万英镑战略设备奖的支持，该研究中心提供了一个相关的桥梁，以联合收割机结合纳米尺度上的结构，功能和成分信息。

Progression of prostate cancer into a metatastic state is postulated to be via exomes, whereby exosomes act to modulate tissues of the body to prepare them to receive and support cancer cell growth. However, addressing this hypothesis has proven challenging as there are many forms of extracellular vesicles of similar size (<150nm) and when isolating extracellular vesicles experimentally at heterogenous population which includes exosomes is collected. Therefore, determination of an enriched exosome fraction prior to experimentation is essential, this requires structural analysis (via cryoTEM) as well as compositional analysis. Through our recent preliminary studies, we can now correlate fluorescently-tagged exosomes optically with structural scanning via cryoTEM. Whilst providing significant structural information, we would like to expand this technique to provide a full compositional analysis at the single exosomes level, this is the first aim of the project. The second aim is to determine the entrance into tissues the exosomes take. This requires a mechanistic and biophysical understanding that is currently lacking. This project will test a selection of different novel workflow methods based around UoN's new cryogenic preparation facility including a plethora of untried correlative workflows to achieve the scientific goals. The project is underpinned by the latest £1.5M strategic equipment award in the Microscale and Nanoscale Research Centre that provides a corelative bridge to combine structural, functional and compositional information on the nanoscale.