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

摘要 通过有针对性和精确的干扰来操纵细胞活动的能力有望极大地 提高我们对生物系统的认识，从单分子到系统水平的细胞生物学。相反的 对于最近用于电子和化学信号控制的光遗传模块的爆炸性增长，没有微扰工具 到目前为止，已经提出了允许对机械信号进行精确的时空控制，尽管 机械信号在许多发育、生理和病理过程中的重要性。这个 开发机械信号扰动工具包的挑战源于这样一个事实，即许多 机械激活的过程在空间和时间上是局部性的，另外还需要机械加载 变得完全激活。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，该系统具有 集成了瞄准、成像和力量生成组件。通过利用这种多功能 纳米探针能力，我们将系统地研究生化相互作用的不同影响 细胞表面、空间受体分离和机械刺激对机械信号的调节 过程和细胞反应。作为初步研究，我们建议研究相互作用和信号传递 神经连接蛋白、整合素和E-钙粘附素在突触功能、细胞基质中的动态变化 相互作用和细胞-细胞连接。最终，我们的目标是为 系统研究各种机械敏感蛋白的工作原理，加快我们的 了解机械信号转导机制和调控。

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.