Elucidating and Directing Heteromolecular Mechanobiology with Nanoengineered Cell Interfaces

项目摘要/摘要 尽管最近在机械生物学研究方面取得了进展，但在关键问题上仍然存在基本的知识差距 细胞功能和分子机制，发生在分子和细胞尺度之间。为 在系统的、定量的和确定性的研究中，实现单分子的精确度是关键的挑战， 杂分子控制，多个物理化学信号的同时和独立调制。基于 我们独特的方法结合了自上而下的纳米技术和自下而上的合成生物学，我们将克服 这些挑战，并创造了“智能”细胞接口，可以探测、感应和操纵细胞和 微环境，以无与伦比的精确和可控的方式。我的实验室的首要目标是 将分子洞察力引入细胞生物学，推进细胞作为机器的知识，其行为 并且功能可以被预测和指导。 基于我们在分子机械生物学几何支撑方面的发现，我们将讨论三个问题 悬而未决的问题：(I)受体聚集的空间效应的基础是什么，它们是如何迫使- 调解？(Ii)在不同受体的共聚和分离中是否存在杂分子空间效应， 它们在信号串扰和整合中扮演什么角色？(三)如何运用分子科学中的新知识 机制和有效地指导细胞功能？考虑到多个信号之间的相互作用和串扰 受体和物理化学线索，我们将揭示杂大分子潜在的协同机制 细胞-细胞外基质(ECM)信号(以成纤维细胞为模型)和细胞-细胞信号(T细胞)的机制生物学 作为模特)。具体地说，对于细胞外基质信号，我们将研究整合素聚集机制，其 与细胞收缩的关系，以及生长因子受体和整合素之间的串扰。对于单元格- 细胞旁分泌信号，我们将研究机械传感和空间传感的协同机制。 在T细胞受体聚集和触发、以及异分子协同聚集机制之间具有激活作用 和抑制性受体。最后，我们在这项研究中开发的人造细胞外基质和人造细胞表面将 用于指导体外细胞工程和制造的关键细胞功能，以及体内的潜力 申请。 总体而言，我们的技术将打破机械生物学研究的限制，在2D、刚性、 静态表面，以3D、柔软、动态的方式提供对多个配体的杂分子控制，以及 因此更好地模拟了复杂的体内环境。这一变革性的方法可以定制 并在不同的系统中采用，以扩大我们对基本分子和细胞机制的理解， 并将新知识应用于新的诊断和治疗方法。

PROJECT SUMMARY/ABSTRACT Despite recent progresses in mechanobiology research, there is still a fundamental knowledge gap in critical cellular functions and molecular mechanisms, which take place between the molecular and cellular scale. For systematic, quantitative and deterministic studies, there are key challenges to achieve single-molecule precision, heteromolecular control, simultaneous and independent modulation of multiple physicochemical cues. Based on our unique approach combining top-down nanotechnology and bottom-up synthetic biology, we will overcome these challenges and create “smart” cell interfaces which could probe, sense and manipulate cells and the microenvironment, in an unparalleled precise and controllable manner. The overarching goal of my laboratory is to bring molecular insights into cell biology, and advance the knowledge of cell as a machine, whose behaviors and functions can be predicted and directed. Based on our discoveries in geometric underpinning of molecular mechanobiology, we will address three outstanding questions: (i) What underlies the spatial effects in receptor clustering, and how are they force- mediated? (ii) Are there heteromolecular spatial effects in co-clustering and segregation of different receptors, and what are their roles in signaling crosstalk and integration? (iii) How to use the new knowledge in molecular mechanisms and effectively direct cell functions? Considering the interplay and crosstalk between multiple receptors and physicochemical cues, we will reveal the underlying synergistic mechanisms of heteromolecular mechanobiology for cell-extracellular matrix (ECM) signaling (fibroblast as a model) and cell-cell signaling (T cell as a model). Specifically, for cell-ECM signaling, we will investigate the integrin clustering mechanism, its relationship with cellular contractility, and the crosstalk between growth factor receptors and integrins. For cell- cell juxtacrine signaling, we will investigate the synergistic mechanism of mechanosensing and spatial sensing in T cell receptor clustering and triggering, and the heteromolecular co-clustering mechanism between activatory and inhibitory receptors. Finally, the artificial ECM and artificial cell surfaces we developed in this research will be used to direct critical cell functions for in vitro cell engineering and manufacturing, as well as potential in vivo applications. Overall, our technology will break the limit of mechanobiology study with a single type of bioligands on 2D, rigid, static surfaces, provide heteromolecular control over multiple ligands in a 3D, soft, dynamic fashion, and therefore better mimic the complex in vivo environment. This transformative methodology can be customized and adopted in distinct systems to expand our understanding of fundamental molecular and cellular mechanisms, and apply the new knowledge for novel diagnostic and therapeutic methods.