Programmed sub-cellular release - a new tool for the study of cell detachment dyn

描述(由申请人提供)：细胞与其细胞外环境的动态附着和脱离调节细胞的运动，对胚胎发育、炎性免疫反应、伤口修复和癌细胞的转移至关重要。这个项目的总体目标是确定细胞收缩、焦点黏附解体和细胞骨架从表面脱离后的全球动力学，并破译驱动和调节这一过程的分子机制。我们的动机是，目前最先进的研究细胞分离涉及诺卡多唑对微管的大量解聚和再聚合，这是一种非特异性的药理学方法，不允许在亚细胞水平上空间控制地释放焦点粘连。技术上的挑战是开发一种工具来研究和控制细胞分离，这种工具将允许在空间和时间上受控的方式在亚细胞区域释放焦点粘连。在初步工作中，我们已经证明了程序化亚细胞释放在解决与细胞分离相关的重要问题方面的可行性。 公共卫生相关性：叙述性细胞分离是从迁移到转移过程的中心。这项拟议的研究将开发一种新的工具来测量细胞脱离，并允许对驱动和调节细胞脱离、焦点黏附分解和细胞骨架重组的分子机制有新的见解。

DESCRIPTION (provided by applicant): The dynamic attachment and detachment of cells to and from their extracellular milieu regulates cell motility and is critical for embryonic development, the inflammatory immune response, wound repair, and metastasis of cancerous cells. The overall goal of this project is to determine the global dynamics of cell contraction, focal adhesion disassembly, and cytoskeleton reorganization after detachment from a surface, and, in addition, to decipher the molecular mechanisms that drives and regulates this process. Our motivation is that the current state-of-the-art to study cell detachment involves bulk depolymerization and repolymerization of microtubules by nocadozole, a non-specific pharmacological approach that does not allow for spatially controlled release of focal adhesions at the sub-cellular level. The technological challenge is to develop a tool to study and control cell detachment that will allow release of focal adhesions over sub-cellular regions in a spatially and temporally controlled manner. In preliminary work we have demonstrated the feasibility of programmed sub-cellular release in addressing important questions related to cell detachment. PUBLIC HEALTH RELEVANCE: Narrative Cell detachment is central to processes from migration to metastasis. The proposed research will develop a new tool to measure cell detachment and allow new insight into the molecular mechanisms that drive and regulate cell detachment, focal adhesion disassembly, and cytoskeleton reorganization.