Project 2: Physical Mechanisms and Clinical Implications of Mechano-transduction

摘要 项目2：肝细胞机械转导的物理机制和临床意义 肿瘤微环境。 在项目2中，来自物理科学、工程学和细胞生物学的研究小组将进行互动 通过临床核心密切联系肝病专家和肝脏肿瘤学家，通过理论核心密切联系理论家。 我们将提出并验证一个新的关于肝细胞癌中机械转导的假说。 微环境。我们假设整个膜信号小体将翻译 物理微环境在膜介导的调节过程中的变化，如受体 运输和膜-皮质相互作用。这反过来又会改变信号通路的特异性，并 影响细胞的命运。理论/膜模型将推进关于物理机制如何支配的假设 生物(细胞)行为，并将指导可在实验中调整的物理参数的设计。超级 分辨率显微镜将被用来跟踪纳米级的组装，力光谱和微观流变学 将用于确定细胞膜和膜皮质相互作用的静态/动态响应。在……里面 平行的高维基因组图谱和单细胞基因表达将把这些纳米级的 与细胞决策有关的机制。这些实验的结果将从数量和机械上 将肝细胞癌中的物理微环境与肿瘤进展中细胞命运的口述联系起来，以及提供 迭代反馈到计算模型中进行机构细化，提出新的假设。 具体地说，项目2的目标将建立物理和物理之间的定量和机械关系 肝细胞癌微环境的特征(即膜张力、基质硬度、基质硬度、 和单轴压应力)和膜介导的信号机制，即如何受体 转运和膜皮质相互作用改变生长因子和G蛋白下游的特异性 调节基因表达和细胞命运的中介信号。我们在项目2中的研究也将探索如何静态 及正常肝细胞膜和膜-皮质相互作用的动态反应 间质细胞会因与肝细胞癌相关的物理微环境变量的变化而改变。我们的项目 在细胞水平上对肝癌的机械转导的研究与项目1的目标密切相关，即 肝细胞癌疾病在组织尺度上的进展，以及项目3的进展，即核力学和肝细胞癌 在亚细胞水平上的肿瘤发生。我们预计，管理肝癌的新物理化学范式 该项目的出现将为未来的肝细胞癌治疗提供信息并产生影响。特别是，我们的结果提供了 亚细胞(膜、皮质、信号、基因表达)的多维、多物理特征 响应微环境变量变化的变化，其中一些是单细胞分辨率的。这个 项目2中的结果是对项目1中的结果的补充，但将细胞的分析和结果扩展到 亚细胞尺度，分子尺度，并帮助在这个更精细的长度尺度上识别物理生物标记物。金诺姆 项目2中的侧写也应该将我们的身体视角与治疗靶点的可能组合联系起来。

Abstract Project 2: Physical Mechanisms and Clinical Implications of Mechano-transduction in Hepatocellular Carcinoma Tumor Microenvironment. In Project 2, a team of investigators from the physical sciences, engineering, and cell biology will interact closely with hepatologists and liver oncologists through the clinical-core, and theorists through the theory-core. We will advance and test a new hypothesis for mechano-transduction in the hepatocellular carcinoma (HCC) microenvironment. We hypothesize that an entire membrane signalosome will translate changes in the physical microenvironment into alterations in membrane-mediated regulatory processes such as receptor trafficking and membrane-cortex interactions. This in turn will alter the specificity of signaling pathways and influence cell fate. Theory/membrane modeling will advance hypotheses on how physical mechanisms govern biological (cellular) behavior, and will direct design of physical parameters tunable in experiments. Super resolution microscopy will be used to track nanoscale assemblies, and force spectroscopy and microrheology will be used to determine static/dynamic responses of the cell membrane and membrane cortex interactions. In parallel, high-dimensional kinome profiling and single-cell gene expression will link these nanoscopic mechanisms with cellular decisions. Outcomes of these experiments will quantitatively and mechanistically relate the physical microenvironment in HCC to dictation of cell fate in cancer progression, as well as providing iterative feedback to the computational models for refinement of mechanisms, formulate new hypotheses. Specifically, the Aims of project 2 will establish quantitative and mechanistic relationships between the physical characteristics of the HCC microenvironment (namely membrane tension, matrix stiffness, substrate stiffness, and uniaxial compressive stress) and membrane-mediated signaling mechanisms, namely how receptor trafficking and membrane cortex interactions alter specificity of downstream of growth factor and G-protein mediated signals to regulate gene expression and cell fate. Our studies in Project 2 will also probe how static and dynamic responses of the cell membrane and membrane-cortex interactions in normal hepatocytes and stromal cells are altered by changes in the physical microenvironment variables relevant for HCC. Our project on mechano-transduction in HCC at the cellular scale is closely aligned with the goals of Project 1, namely HCC disease progression at the tissue scale, and those of Project 3, namely nuclear mechanics and HCC oncogenesis at the subcellular scale. We expect that the new physical-chemical paradigms governing HCC emerging from this project will inform and impact future HCC therapies. In particular, our results provide multidimensional, multiphysics characterization of subcellular (membrane, cortex, signals, gene-expression) alterations in response to changes in the microenvironment variables, some at single-cell resolution. The findings in Project 2 compliment those in Project 1, but extend the analyses and outcomes at the cellular to sub-cellular scale, molecular scale, and help identify physical biomarkers at this finer length-scale. Kinome profiling in Project 2 should also link our physical perspective to possible combinations of therapeutic targets.