Probing the mechano-biology of cell-cell adhesion in a novel single cell assay

我们、动物和许多其他生物都是由许多细胞构成的。它们不是被动地粘在一起的简单积木，而是每个细胞都包含一种复杂的机制来连接邻近的细胞并相互施加力。这种至关重要的能力是由细胞表面的多个粘附分子提供的，这些粘附分子通过适配蛋白连接到产生力的细胞骨架上。在发育过程中，力量的平衡以及不同细胞相互粘附的强度都是细胞运动和分化协调的重要线索。现在很清楚，细胞水平上的机械变化可以改变组织的命运。在癌症中，细胞的脱离导致转移与细胞力学的改变有关。我们有兴趣剖析细胞内机械信号在细胞-细胞粘附的基础。为此，我们正在建立一个新的国际合作，以研究力量在细胞粘附过程中的作用。我们用一种新颖的方法用高分辨率荧光显微镜观察细胞-细胞粘附界面，用含有粘附分子的平面脂质双分子层代替一个细胞。这将结合三种模式的机械操作：约束，剪切应力和横向拉伸。除了荧光显微镜技术跟踪粘附和细胞骨架蛋白的动态，我们将采用干涉反射显微镜以纳米精度揭示细胞膜的波动。这将使我们对细胞粘附过程中的蛋白质动力学和细胞膜力学有一个前所未有的了解。用于此类实验的工具通常由单个实验室建造，这限制了人们使用此类设备的范围。这就是为什么我们想利用这次合作的综合专业知识来设计和制造MechanoWOSM，这是一个用于机械生物学实验的开源显微镜平台。

We, animals and many other organisms are built from lots of cells. These are not simple building blocks that are passively stuck together, but each cell contains an elaborate machinery to connect to its neighbours and exert force on one other. This crucial ability is provided by multiple adhesion molecules at the cell surface that link up to the force generating cytoskeleton via adapter proteins. The balancing of forces and how strong different cells stick to each other are all important cues in the orchestration of cell motion and differentiation during development. And it is now clear that mechanical changes on the cellular level can alter the fate of the tissue. In cancer, the detachment of cells leading to metastasis is related to a change of cell mechanics.We are interested to dissect the basis of intracellular mechanical signalling in cell-cell adhesion. For this, we are establishing a new international collaboration to study the role of forces in the process of cell adhesion. We use novel assay to look at the cell-cell adhesion interface with high resolution fluorescence microscopy by replacing one cell with a planar lipid bilayer containing adhesion molecules. This will be combined with three modes of mechanical manipulations: confinement, shear stress and lateral stretch. In addition to fluorescence microscopy techniques to follow the dynamics of adhesion and cytoskeletal proteins, we will employ interferometric reflection microscopy to reveal the fluctuations of the cell membrane with nanometre precision. This will give us an unprecedented insight into the protein dynamics and cell membrane mechanics during cell adhesion. The tools for such experiments are often built by individual labs which limits the range of people having access to such equipment. That's why we want to use the combined expertise of this collaboration to design and engineer MechanoWOSM, an open source microscope platform for mechano-biology experiments.