Fundamental Mechano-Chemical Mechanisms of Signaling in Cancer

这项提议的长期目标是发展分子水平的理解，了解机械力可以对化学信号过程施加调控。我们寻求对细胞的机械环境如何影响其细胞内化学信号的基本了解。为了实现这一目标，我们提出了一种高度多学科的、混合的物理和生物学方法，旨在解构在癌症中重要的关键化学信号转导通路可以响应机械输入。其基本原理是，信号蛋白质的空间组织在导致恶性肿瘤的不同阶段发生改变，这从根本上讲是蛋白质结构的化学突变，而是大分子长度尺度上的蛋白质组织的物理扰动。我们方法的前提是，表征和控制驱动受体组织的机械力将允许我们在癌症进展的特定阶段诱导出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。它们之所以被选中，是因为它们在化学机械信号转导中的新角色。我们将实施一种组合方法，包括1)混合细胞支持的膜连接的超分辨率成像，2)基于微悬臂的配基功能化探针的侧向力测量，以及3)用于细胞骨架破坏的纳米级激光手术。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，该策略提供了机械干扰具有化学特异性的活细胞的独特机会。数学建模是所有定量研究中必不可少的一部分，在这里也是如此。 由于缺乏方法学，这里所研究的化学机械信号转导偶联的类型在很大程度上被更经典的生物学方法所忽视。这个项目非常适合这个中心，它是建立在物理和生物方法的杂交中的。

The long-term goal of this proposal is to develop a molecular level understanding of hew mechanical forces can exert regulatory control ever chemical signaling processes. We seek a fundamental understanding of how the mechanical environment of a cell influences its intracellular chemical signaling. To achieve this goal we propose a highly multidisciplinary, hybrid physical and biological approach aimed at deconstructing hew 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 to malignancy, and this is fundamentally net a chemical mutation in the structure of a protein, but rather a physical perturbation to protein organization en the macromolecular length scale. 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 in defined phases of 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 to mechanically perturb living cells with chemical specificity. Mathematical modeling is an essential part of all quantitative investigations and is integrated here as well. The types of chemomechanical signal transduction couplings under investigation here have been largely overlooked by more classical biological approaches because of lack of methodology. This project is ideally suited to this center, which is built in the hybridization of physical and biological approaches.