Spatiotemporal interrogation of molecular mechanobiololgy at the cell-cell interface with nanotechnology tools

摘要 Juxtacrine信号传导通过信号传导处的直接分子相互作用介导细胞-细胞通信 界面，在发育过程中，突触形成和重塑，免疫活动和组织形成。 尽管越来越多的知识，这些信号事件，很少有人知道如何阿托克林受体 感知并调节细胞信号以响应其周围细胞的动态变化。的挑战 询问阿曲他克林细胞-细胞信号传导的时空动力学源于以下事实： 受体整合化学、空间和机械信号以差异地调节细胞信号传导。到 解构和解码这些具有高度时空复杂性的受体的工作机制， 技术工具，允许操纵具有不同刺激模式的个体线索， 以高时空精度报告细胞反应。为了实现这一目标，我们以前开发了 纳米技术平台，包括单价量子点（mQD）探针、机械遗传学、纳米粒子 力显微镜（NRFM）和磁放大蛋白质-蛋白质相互作用（MAP-I）工具。mQD报告 靶向受体的单分子轨迹，提供其动态空间和扩散特性 没错机械遗传学使我们能够操纵的化学，空间和机械性能， 靶向受体，同时监测细胞对相应线索的反应。NRFM使我们能够调查 力响应的结构变化的目标受体，因此提供了重要的见解， 机械传导机制MAP-I允许超灵敏地检测蛋白质-蛋白质相互作用 通过磁放大，能够识别尚未被发现的弱蛋白质-蛋白质相互作用， 任何其他技术都可能。通过使用这些新技术，在这里，我们建议调查 Notch和Neuroligin的相互作用和信号动力学，发育中的关键信号蛋白， 突触功能。最终，我们的目标是提供一个平台技术， 研究广泛的阿曲他克林信号传导的操作原理，加速我们对 细胞间通讯

Abstract Juxtacrine signaling mediates cell-cell communications via direct molecular interactions at the signaling interface, during development, synapse formation and remodeling, immune activities, and tissue formation. Despite increasing knowledge of these signaling events, little is known about how the juxtacrine receptors sense and regulate cell signaling in response to the dynamic changes of its surrounding cells. The challenge of interrogating spatiotemporal dynamics of juxtacrine cell-cell signaling stems from the fact that many juxtacrine receptors integrate chemical, spatial, and mechanical cues to differentially regulate cell signaling. To deconstruct and decode the working mechanisms of these receptors with high spatiotemporal complexity, new technology tools allowing manipulation of the individual cues with different modes of stimulation, while reporting cellular responses with high spatiotemporal precision. Toward this aim, we previously developed nanotechnology platforms including monovalent quantum dot (mQD) probes, mechanogenetics, nanoruler force microscopy (NRFM), and magnetically amplified protein-protein interaction (MAP-I) tools. mQDs report single molecule trajectories of the targeted receptors, providing its dynamic spatial and diffusion properties precisely. Mechanogenetics allows us to manipulate chemical, spatial, and mechanical properties of the targeted receptors, while monitoring cellular responses to the respective cues. NRFM enables us to investigate force-responsive structural changes of the target receptors, and hence provides important insights into the mechanism of mechanotransduction. MAP-I allows for ultrasensitive detection of protein-protein interactions through magnetic amplification, enabling identification of weak protein-protein interactions that have not been possible with any other technologies. By using these new technologies, here, we propose to investigate the interaction and signaling dynamics of Notch and Neuroligin, key signaling proteins in development and synaptic function, respectively. Ultimately, we aim to provide a platform technology for the systematic investigation of operating principles for a wide range of juxtacrine signaling, accelerating our understanding of cell-cell communication.