Decoding cortical Notch signaling and morphogenic instruction at cell-cell interfaces

项目总结/摘要 遗传程序的协调和细胞的物理组织塑造了发育中的组织， 再生，当失调时，促进疾病。这两个过程在空间上是如何协调的， 人们对时间知之甚少。细胞-细胞界面是形态发生指令的重要组织中心。这里 影响最终决定细胞命运的基因表达的信号与细胞周期的变化相结合。 力学和，在一起，是必要的，以建立和维持多细胞架构。升值幅度 基因调控网络和机制在细胞-细胞界面共同指导形态发生过程， 一个新出现的挑战是通过实验来确定它们在各种不同情况下的交叉操作的细节， 形态发生背景细胞间通讯的一个有趣的例子是无处不在的重要缺口 受体途径具有调节细胞力学和基因表达的内在能力，但 这些不同活动的机制尚不清楚。作为博士后研究员，PI开发了仿生 人体组织的微流体模型，用于确定控制3D的几种新机制 在细胞-细胞和细胞-细胞外基质界面上操作的多细胞行为。PI发现， 高度保守的Notch受体家族具有以前未描述的皮质信号传导功能， 允许Notch将细胞力学的变化与转录输出联系起来。作为一个独立的实验室， Kutys实验室已经完成了对上皮组织中皮质Notch信号传导的首次研究， 形态发生的后果和以前不受重视的信号传导机制。在接下来的五年里， Kutys实验室的目标是继续其开发下一代仿生的多学科方法 人体组织系统，分子技术和基于显微镜的方法来确定协调 细胞粘附界面的形态发生行为和信号传导。我们会集中在三个范畴： 设计新工具，深入了解皮质Notch通路如何调节信号传导和粘附 上皮细胞中的机制及其被调用以调节体外重要生物过程的背景 2）全面定义Notch受体的定位和激活如何受到以下因素的影响： 在上皮细胞拥挤和内皮细胞增生过程中与细胞间粘附和皮质肌动蛋白的生物物理相互作用 细胞暴露于剪切应力，和3）整合蛋白质组学方法与3D仿生系统，以广泛 剖析和剖析组织结构如何影响细胞-细胞界面上的分子控制系统。 这项研究的结果和使能技术的整合将有助于整体 我的研究计划的目标。总之，这些研究将提供一个重要的新的基本见解 Notch信号传导臂，细胞-细胞粘附如何动态调节，以及如何调节的分子基础。 转录和粘附程序可能在复杂的3D组织内协调。

Project Summary/Abstract The coordination of genetic programs and the physical organization of cells sculpt developing tissues, drive regeneration and, when dysregulated, facilitate disease. How these two processes are coordinated in space and time is poorly understood. Cell-cell interfaces are important organizing centers for morphogenic instruction. Here signals influencing gene expression that ultimately dictate cell fate decisions are integrated with changes in cell mechanics and, together, are necessary to build and maintain multicellular architectures. With an appreciation that gene regulatory networks and mechanics conspire at cell-cell interfaces to instruct morphogenic processes, an emerging challenge is to experimentally define the details of their intersectional operations in diverse morphogenic contexts. One intriguing example of cell-cell communication is the ubiquitously important Notch receptor pathway which has the intrinsic capacity to regulate both cell mechanics and gene expression, yet mechanisms of these distinct activities are unclear. As a postdoctoral fellow, the PI developed biomimetic microfluidic models of human tissues that were employed to identify several new mechanisms controlling 3D multicellular behavior operating at cell-cell and cell-extracellular matrix interfaces. The PI discovered that the highly conserved Notch family of receptors possess a previously undescribed cortical signaling function that permits Notch to connect changes in cell mechanics to transcriptional output. As an independent laboratory, the Kutys Lab has completed the first investigation into cortical Notch signaling in epithelial tissues and has identified morphogenic consequences and previously unappreciated signaling mechanisms. Over the next five years, the goal of the Kutys Lab is to continue its multidisciplinary approach of developing next generation biomimetic human tissue systems, molecular technologies, and microscopy-based methods to define the coordination of morphogenic behavior and signaling at cell adhesive interfaces. We will focus these efforts in three Areas: 1) engineering new tools to deeply understand how the cortical Notch pathway regulates signaling and adhesion mechanics in epithelia and the contexts in which it is invoked to regulate important biological processes in vitro and in vivo, 2) comprehensively defining how Notch receptor localization and activation are influenced by biophysical interactions with cell-cell adhesions and cortical actin during epithelial crowding and in endothelial cells exposed to shear stress, and 3) integrating proteomic approaches with 3D biomimetic systems to broadly profile and dissect how tissue architecture influences molecular control systems operating at cell-cell interfaces. The results of this research and the integration of the enabling technologies will contribute to the overall objectives of my research program. Together, these studies will offer fundamental insight into an important new arm of Notch signaling, how cell-cell adhesions might be dynamically regulated, and the molecular basis for how transcriptional and adhesive programs might be coordinated within complex 3D tissues.