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

在新型单细胞测定中探讨细胞间粘附的力学生物学

• 批准号: EP/Y002245/1
• 负责人: Darius Koester
• 金额: $ 21.07万
• 依托单位: University of Warwick
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2024
• 资助国家: 英国
• 起止时间: 2024 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=EP%2FY002245%2F1
• 关键词: Probing; mechano; biology; cell; adhesion

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.

我们、动物和许多其他有机体都是由大量细胞构成的。这些并不是被动地粘在一起的简单构建块，而是每个细胞都包含一个复杂的机制来连接到相邻的细胞并向彼此施加力。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，这些粘附分子通过衔接蛋白连接到产生力的细胞骨架。力的平衡以及不同细胞彼此粘附的强度都是发育过程中细胞运动和分化协调的重要线索。现在很明显，细胞水平上的机械变化可以改变组织的命运。在癌症中，导致转移的细胞脱离与细胞力学的变化有关。我们有兴趣剖析细胞间粘附中细胞内机械信号传导的基础。为此，我们正在建立一个新的国际合作来研究力在细胞粘附过程中的作用。我们使用新颖的测定法，通过用含有粘附分子的平面脂质双层替换一个细胞，用高分辨率荧光显微镜观察细胞-细胞粘附界面。这将与三种机械操作模式相结合：限制、剪切应力和横向拉伸。除了荧光显微镜技术来跟踪粘附和细胞骨架蛋白的动态之外，我们还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前所未有的了解。用于此类实验的工具通常是由各个实验室构建的，这限制了使用此类设备的人员范围。这就是为什么我们希望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。