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

使用纳米技术工具对细胞-细胞界面处的分子力学生物学进行时空询问

基本信息

批准号：
10359739
负责人：
Young-wook Jun
金额：
$ 53.54万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-03-01 至 2025-02-28
项目状态：
未结题

项目摘要

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.

抽象的近距离信号传导通过信号传导下的直接分子相互作用介导细胞细胞通信界面在发育，突触形成和重塑，免疫活性和组织形成。尽管对这些信号事件的了解越来越多，但对近距离受体的了解知之甚少响应其周围细胞的动态变化，将其感知和调节细胞信号传导。挑战近距离细胞 - 细胞信号传导的询问时空动力学源于许多近去氨酸受体将化学，空间和机械提示整合到差异调节细胞信号传导中。到解构和解码这些受体具有高时空复杂性的工作机制，新的技术工具允许以不同的刺激方式操纵单个线索，而报告具有高时空精度的细胞反应。为了实现这一目标，我们以前开发了纳米技术平台在内力显微镜（NRFM）和磁化的蛋白质 - 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恰恰。机械遗传学使我们能够操纵化学，空间和机械性能靶向受体，同时监测对各自提示的细胞反应。 NRFM使我们能够调查力响应性的目标受体的结构变化，因此提供了重要的见解机理的机理。 MAP-I允许对蛋白质 - 蛋白质相互作用的超敏检测通过磁扩增，可以鉴定弱蛋白质 - 蛋白质相互作用使用任何其他技术。通过使用这些新技术，我们建议调查 Notch和Neuroligin的相互作用和信号动力学，开发中的关键信号蛋白和突触功能分别。最终，我们旨在为系统性提供平台技术对多种近二氨酸信号传导的操作原理的调查，加速了我们对细胞电池通信。