Nanomodules for interrogating chemical, spatial, and mechanical dynamics of cell surface receptors

摘要 通过有针对性的和精确的扰动来操纵细胞活动的能力有望显着提高 增强我们对生物系统的理解，从单分子到系统水平的细胞生物学。相反 到最近用于电和化学信号控制的光遗传学模块的爆炸，没有微扰工具 迄今为止，已经提出了允许机械信号的精确时空控制的方法，尽管 机械信号在许多发育、生理和病理过程中的重要性。的 开发用于机械信号传导的扰动工具包的挑战源于以下事实： 机械激活过程在空间和时间上是局部的，并且另外需要机械加载 完全激活。为了解决这个问题，我们建议开发一种先进的纳米探针系统， 整合了瞄准、成像和力量生成组件。通过利用这种多功能 纳米探针的能力，我们将系统地研究生化相互作用的差异影响， 细胞表面、空间受体分离和机械刺激对机械信号转导的调节 过程和细胞反应。作为初步研究，我们建议调查相互作用和信号 神经连接素、整合素和E-钙粘蛋白的动力学，突触功能中的关键信号蛋白，细胞基质 相互作用和细胞-细胞连接。最终，我们的目标是提供一个平台技术， 系统地研究了各种机械敏感蛋白的工作原理，加速了我们的研究。 了解机械信号机制和调节。

ABSTRACT The ability to manipulate cellular activities through targeted and precise perturbation promises to dramatically enhance our understanding of biological systems from single molecules to systems-level cell biology. Contrary to the recent explosion of optogenetic modules for electro and chemical signal control, no perturbative tools allowing precise spatiotemporal control of mechanosignaling have been presented so far, despite the importance of mechanosignaling in many developmental, physiological, and pathological processes. The challenge of developing a perturbation toolkit for mechanosignaling stems from the fact that many mechanically-activated processes are localized in space and time and additionally require mechanical loading to become fully activated. To address this, we propose to develop an advanced nanoprobe system with integrated targeting, imaging, and force-generating components. By taking advantage of such multifunctional nanoprobe capabilities, we will systematically investigate the differential effects of biochemical interaction at cell surface, spatial receptor segregation, and mechanical stimulation on regulation of mechanosignaling processes and cellular responses. As initial studies, we propose to investigate interaction and signaling dynamics of neuroligin, integrin, and E-cadherin, key signaling proteins in synaptic function, cell-matrix interactions, and cell-cell junctions, respectively. Ultimately, we aim to provide a platform technology for the systematic investigation of operating principles for a wide range of mechanosensitive proteins, accelerating our understanding of mechanosignaling mechanisms and regulation.