Developing NMR Methods to Characterise Cell Signalling Pathways for Novel Drug Discovery

细胞检测并响应其环境的局部机械特性。细胞周围环境的粘度和硬度的变化会导致细胞功能和代谢的显著变化，但这些细胞变化的机制仍然知之甚少。然而，这些效应是非常重要的;例如，随着组织硬度的增加，癌细胞对周围组织的侵袭得到促进。在这一领域的研究中的一个障碍是缺乏方法来测量细胞周围的刚度和粘度的局部变化。除此之外，还需要评估向细胞发出信号的细胞外基质分子的结构和动力学如何受到局部刚度/粘度变化的影响的方法，以了解机械性质变化对来自细胞外环境的细胞信号传导的分子效应。该项目将首先开发溶液状态、凝胶状态和固态NMR方法，以测量细胞周围局部粘度和刚度的变化。然后，我们将开发一个NMR工具包，以了解细胞外基质分子的动力学和构象如何与基质粘度和刚度相关。最后，我们将使用核磁共振方法和光学成像来了解细胞外基质分子与细胞的界面，以产生细胞信号。将整个项目的结果汇总在一起，将导致关于细胞如何感知环境的新假设。总的来说，该项目将提供物理方法，以加强化学和生物学之间的研究。

Cells detect and respond to the local mechanical properties of their environment. Changes in the viscosity and stiffness of a cell's surroundings cause significant changes in cell function and metabolism, but the mechanisms for these cell changes are still poorly understood. These effects are however very important; cancer cell invasion of surrounding tissue is promoted with increasing tissue stiffness, for instance. One of the blocks in research in this area is the paucity of methods to measure local changes in stiffness and viscosity around cells. Alongside this is the need for methods to assess how the structure and dynamics of extracellular matrix molecules that signal to cells are affected by changes in local stiffness/ viscosity, in order to understand the molecular effect of mechanical property changes on cell signalling from the extracellular environment. This project will first develop solution-state, gel-state and solid-state NMR methods to measure changes in local viscosity and stiffness around cells. We will then progress to develop an NMR toolkit to understand how the dynamics and conformations of extracellular matrix molecules correlate with the matrix viscosity and stiffness. Finally, we will use NMR methods and optical imaging to understand of extracellular matrix molecues interface with cells to generate cell signaling. Drawing together the results from the whole project will lead to new hypotheses about how cells sense their environment. Overall, the project will provide physical methods to enhance research at the interface between chemistry and biology.