Integrative Approaches for Probing Cell Mechanotransduction in Health and Disease

项目摘要 Notch受体是一种机械激活的跨膜蛋白，在调节细胞中发挥重要作用。 命运、分化、增殖、黏附等许多关键过程。然而，大多数研究都是 到目前为止，主要集中在从生化和遗传学的角度理解Notch，而且只有 最近被认为是一种机械感受器。鉴于Notch在正常和病理中扮演的不同角色 国家，有必要制定一个完整的机械力化学观点的Notch信号和调控 因此，要全面了解这一关键途径，就需要有这样的观点。在这个项目中，我的实验室将 利用我们在化学探针开发和分子工具设计方面的专业知识，以大幅解决 关于Notch机械转导的重点机械问题。为了区别我们的贡献 从其他人的研究中，我们将通过结合单个分子来寻求对该途径的多尺度理解 研究和高分辨率成像，目的是了解受体的激活是如何耦合的 细胞内发生的生物力学事件。特别是，新的相关光学和电子显微镜 将被开发和应用来确定围绕陷波信号的事件的精确定时和位置 转导。此外，为了解决有关资金来源和规模的重要问题， 细胞所经历的外力，我们还将创建Notch的合成版本并将其应用于 基因编码为“张力计”。这项工作的成功实施将为协同效应提供深刻的见解 在生化和机械提示之间发生，以及增加我们对如何 细胞感知和解释机械信息。

Project Summary Notch receptors are mechanically activated transmembrane proteins that play important roles in regulating cell fate, differentiation, proliferation, adhesion, and many other critical processes. However, the majority of studies to date have been focused largely on understanding Notch from biochemical and genetic perspectives, and only recently been explored as a mechanoreceptor. Given the varied roles Notch plays in both normal and pathological states, it is necessary to formulate an integrated mechano-chemical perspective of Notch signaling and regulation as such outlook is required to achieve a comprehensive view of this critical pathway. In this project, my lab will leverage our expertise in chemical probe development and molecular tool design in order to address sharply focused mechanistic questions regarding Notch mechanotransduction. In order to distinguish our contributions from those of others, we will pursue a multi-scale understanding of the pathway by combining single molecule studies and high-resolution imaging with the aim of understanding how the activation of the receptor is coupled to biomechanical events that occur within the cell. In particular, new correlative light and electron microscopy will be developed and applied to determine the precise timing and location of events surrounding Notch signal transduction. In addition, in order to address important questions regarding the source and magnitude of the external forces that are experienced by cells, we will also create synthetic versions of Notch and apply them as genetically encoded “tensiometers.” Successful execution of this work will provide deep insights into the synergy occurring between biochemical and mechanical cues as well as increase our overall understanding of the how cells sense and interpret mechanical information.