Modeling bi-directional signaling and cytoskeletal dynamics in 3D cell migrations

描述(由申请人提供)：在健康和疾病的系统中，细胞的结构和功能由细胞与潜在和周围的三维细胞外基质的相互作用调节。这些复杂的生化和生物力学相互作用，独立地调节肿瘤的进展、侵袭和转移是众所周知的。例如，转移性乳腺癌中细胞对生化和生物物理刺激的异常反应通常是由细胞骨架机械的参与启动的。因此，肌动蛋白相互作用的蛋白质在生化信号和促进黏附信号之间的信号网络串扰的连接点上被发现。一个这样的例子是MENA，它是肌动蛋白调节蛋白Ena/Vasp家族的成员，已经被描述为在侵袭和转移过程中异常的细胞信号反应。然而，改变后的信号网络如何转化为机械过程，以及这些亚细胞机械过程如何转化为3D环境中的整体细胞迁移，在很大程度上仍然难以捉摸。在这里，基于我们的初步数据，我们假设3D环境中肿瘤细胞侵袭力的增加是由分子水平的异常信号事件与分子、大分子和细胞生物力学过程耦合所控制的。我们在这项建议中的主要目标是通过弥合分子水平的体外信号研究与3D的分子力学和细胞模型之间的知识差距来严格检验我们的假设，并通过3D环境中的定量实验来测试我们的模型的预测。我们计划使用以下三个具体目标来开发和验证我们的细胞模型：目标一：开发细胞骨架粘弹性和细胞内信号传递的集成亚细胞模型 在本地的类似3D矩阵中。目的II：在3D矩阵中建立细胞迁移的定量模型， 利用AIM I的亚细胞模型的结果。AIM III：验证AIM I和II B的结果，量化信号如何与细胞力学机制和细胞外基质属性协同作用，以3D方式调节细胞迁移。这三个目标都建立在计算和实验研究的强大初步数据基础上，并将提供对机械和生化途径之间的耦合以及从亚细胞结构到细胞水平的信息整合的基本见解。同时，对3D环境的关注将创造关于类似本地环境中的蜂窝系统的新的和生理相关的知识。最后，通过这项工作开发的新平台将能够测试临床相关的假说，并有助于定量理解癌症进展不同阶段的复杂多尺度过程。

DESCRIPTION (provided by applicant): Cellular structure and function, in healthy and diseased systems, is regulated by the interaction of cells with the underlying and surrounding three-dimensional extra-cellular matrix. These complex biochemical and biomechanical interactions, independently, are well known to regulate tumor progression, invasion and metastasis. For example, the aberrant response of cells to biochemical and biophysical stimuli in metastatic breast cancer is often initiated by engagement of the cytoskeletal machinery. As such, actin interacting proteins are found at the nexus of signaling network crosstalk between biochemical and adhesion-promoting cues. One such example is Mena, a member of the Ena/VASP family of actin regulatory proteins, which has been characterized for aberrant cell-signaling response during invasion and metastasis. However, how the altered signaling network is translated into the mechanical processes, and how are these sub-cellular mechanical processes then converted into whole cell migration in 3D environments remain largely elusive. Here, based on our preliminary data, we hypothesize that increased tumor cell invasiveness in 3D environments, is governed by coupling aberrant molecular level signaling events to molecular, macromolecular and cellular biomechanical processes. Our primary goal in this proposal is to rigorously test our hypothesis by bridging the knowledge gap between in vitro signaling studies at the molecular level, and molecular mechanical and cellular models in 3D, and test the predictions of our models through quantitative experiments in 3D environments. We plan to develop and validate our cellular models using the following three specific aims: Aim I: Develop an integrated subcellular model of cytoskeletal viscoelasticity and intracellular signaling in native like 3D matrices. Aim II: Develop a quantitative model of cell migration, in 3D matrices, utilizing results from the subcellular model of Aim I. Aim III: Validate results of Aims I and II b quantifying how signaling acts cooperatively with cellular mechanics machinery and extracellular matrix properties to regulate cell migration in 3D. All three aims build upon strong preliminary data in both computation and experimental studies and will provide both fundamental insights into the coupling between mechanical and biochemical pathways and integration of information from sub-cellular structures to the cellular level. At the same time, the focus on 3D environments will create new and physiologically relevant knowledge about cellular systems in native like environments. Finally, novel platforms developed through this work will be able to test clinically relevant hypotheses and help in quantitatively understanding complex multi-scale processes during various stages of cancer progression.