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

摘要 辅助性他克林信号通过信号转导的直接分子相互作用介导细胞间的通讯 在发育过程中，突触的形成和重塑、免疫活动和组织形成。 尽管对这些信号事件的了解越来越多，但对旁分泌受体如何 感知和调节细胞信号，以响应周围细胞的动态变化。面临的挑战 质疑旁分泌细胞-细胞信号的时空动力学源于这样一个事实：许多旁分泌 受体整合化学、空间和机械信号，以不同的方式调节细胞信号。至 解构和解码这些具有高时空复杂性的受体的工作机制，新的 技术工具，允许通过不同的刺激模式操纵单独的提示，而 以高时空精确度报告细胞响应。为了达到这个目标，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、纳米规则 力显微镜(NRFM)和磁放大蛋白质-蛋白质相互作用(MAP-I)工具。MQDS报告 靶向受体的单分子轨迹，提供其动态空间和扩散特性 正是如此。机械遗传学允许我们操纵化学、空间和机械特性 靶向受体，同时监测细胞对各个线索的反应。NRFM使我们能够调查 目标受体的力响应结构变化，从而提供了对 机械转导机制。MAP-I允许超灵敏地检测蛋白质-蛋白质相互作用 通过磁性放大，能够识别尚未被 任何其他技术都是可能的。通过使用这些新技术，我们在这里建议调查 发育和发育中的关键信号蛋白Notch和Neuroigin的相互作用和信号动力学 突触功能。最终，我们的目标是为系统提供一个平台技术 研究广泛的旁分泌信号的工作原理，促进我们对 细胞间的通讯。

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.