Correlative Nano-Characterisation for Prostate Cancer

前列腺癌向转移状态的进展被认为是通过外切体，由此外切体作用于调节身体组织，使它们准备接收和支持癌细胞的生长。然而，解决这一假设已被证明具有挑战性，因为有许多形式的细胞外小泡相似的大小(&lt；150 nm)，当在实验中分离细胞外小泡时，收集到包括外体的异质种群。因此，在实验前确定浓缩的外切体组分是必要的，这需要结构分析(通过CryoTEM)和成分分析。通过我们最近的初步研究，我们现在可以将荧光标记的外切体与通过CryoTEM进行的结构扫描进行光学关联。在提供重要的结构信息的同时，我们希望扩展这项技术，以便在单个外体水平上提供完整的成分分析，这是该项目的第一个目标。第二个目标是确定外切体进入组织的途径。这需要对机械和生物物理的理解，而这是目前所缺乏的。该项目将围绕UON的新低温制备设施测试一系列不同的新型工作流方法，包括大量未尝试过的相关工作流，以实现科学目标。该项目得到了微尺度和纳米尺度研究中心最新的GB 150万战略装备奖的支持，该奖项提供了一座相互关联的桥梁，将纳米尺度上的结构、功能和组成信息结合在一起。

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.