Fundamental Mechano-Chemical Mechanisms of Signaling in Cancer

项目I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。这项建议的长期目标是发展分子水平的理解，了解机械力如何对化学信号过程施加调节控制。我们试图从根本上理解细胞的机械环境对其细胞内化学信号的影响。为了实现这一目标，我们提出了高度多学科的研究方法。旨在解构癌症中重要的关键化学信号转导通路如何对机械输入做出反应的混合物理和生物方法。其基本原理是，信号蛋白的空间组织在导致TEE恶性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(7，2)上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得出癌症进展过程中特定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。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，该策略提供了独特的机会，可以机械地干扰具有化学特异性的活细胞。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。

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.