Fundamental Mechano-Chemical Mechanisms of Signaling in Cancer

这项计划的长期目标是在分子水平上理解新的机械力可以对化学信号过程施加调节控制。我们寻求对细胞的机械环境如何影响其细胞内化学信号的基本理解。为了实现这一目标，我们提出了一个高度多学科的，混合物理和生物学的方法，旨在解构新的关键化学信号转导途径的意义，在癌症中可以响应机械输入的基本原理是，信号蛋白的空间组织被改变，在不同的阶段，导致恶性肿瘤，这是根本上净化学突变的蛋白质结构，而是在大分子长度尺度上对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使我们能够在癌症进展的定义阶段中引出结构和功能表型特征。 我们将靶向EphA2受体信号通路以及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）混合细胞支持的膜连接的超分辨率成像，2）基于微悬臂梁的配体功能化探针的侧向力测量，和3）用于细胞骨架破坏的纳米剪刀激光手术。所有这三种方法都将在新开发的空间突变策略的背景下采用，该策略提供了独特的机会，以机械方式干扰具有化学特异性的活细胞。数学建模是所有定量研究的重要组成部分，也是这里的一部分。 由于缺乏方法学，更经典的生物学方法在很大程度上忽视了这里研究的化学机械信号转导偶联的类型。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。

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.