Fundamental Mechano-Chemical Mechanisms of Signaling in Cancer

项目1的中心假设是信号蛋白的空间组织被机械力主动改变，这为物理力直接调节癌症中的化学信号提供了分子尺度的机制。本提案的长期目标是在分子水平上理解机械力如何对化学信号传导过程施加调节控制。我们寻求对细胞的机械环境如何影响其细胞内化学信号传导的基本理解。为了实现这一目标，我们提出了高度多学科化。混合物理和生物方法旨在解构癌症中重要的关键化学信号转导途径如何响应机械输入。其基本原理是信号蛋白的空间组织在导致恶性肿瘤的不同阶段发生改变，这基本上不是蛋白质结构的化学突变，而是大分子长度尺度上蛋白质组织的物理扰动（7,2）。我们方法的前提是表征和控制驱动受体组织的机械力将使我们能够引出癌症进展中定义阶段的结构和功能表型特征。我们将针对EphA2受体信号通路和Ras信号模块。选择这些是因为它们在化学机械信号转导中的新兴作用。我们将实施一种组合方法，包括1)杂交细胞支持的膜连接点的超分辨率成像，2)基于配体功能化探针的微悬臂横向力测量，以及3)纳米剪刀激光手术治疗细胞骨架破坏。所有这三种方法都将在新开发的空间突变策略的背景下使用，该策略提供了独特的机会，可以用化学特异性机械地干扰活细胞。数学建模是所有定量研究的重要组成部分，在这里也得到了整合。

The central hypothesis of Project I is that the spatial organization of signaling proteins is actively altered by mechanical forces and this provides a molecular scale mechanism for physical forces to directly regulate chemical signaling in cancer. The long-term goal of this proposal is tee develop a molecular level understanding of how mechanical forces can exert regulatory control over chemical signaling processes. We seek a fundamental understanding of hew the mechanical environment of a cell influences its intracellular chemical signaling. Te achieve this goal, we propose a highly multidisciplinary. hybrid physical and biological approach aimed at deconstructing how key chemical signal transduction pathways of significance in cancer can be responsive to mechanical inputs. The rationale for this is that the spatial organization of signaling proteins is altered in the different phases leading tee malignancy, and this is fundamentally net a chemical mutation in the structure of a protein, but rather a physical perturbation of protein organization on the macromolecular length scale (7, 2). The premise of our approach is that characterizing and controlling mechanical forces that drive receptor organization will allow us to elicit structural and functional phenotypes characteristic of defined phases of in cancer progression. We will target the EphA2 receptor signaling pathway as well as the Ras signaling module. These are chosen for their emerging roles in chemomechanical signal transduction. We will implement a combined approach that consists of 1) super-resolution imaging of hybrid cell-supported membrane junctions, 2) micro cantilever-based lateral force measurements of ligand-functionalized probes, and 3) nanoscissor laser surgery for cytoskeletal disruption. All three of these approaches will be employed in the context of the newly developed spatial mutation strategy, which provides unique opportunities tee mechanically perturb living cells with chemical specificity. Mathematical modeling is an essential part of all quantitative investigations and is integrated here as well.