Role of vimentin in mammalian cell motility

摘要 细胞迁移是许多基本过程的关键，包括胚胎发育、伤口修复、 和癌症转移。这一过程的中心是细胞细胞骨架，它由三个聚合物组成 网络：F-肌动蛋白、微管和中间丝。波形蛋白是一种IF蛋白，它是 细胞的机械弹性和调节细胞骨架之间的串扰，但它在细胞如何 人们对挤压组织中的小孔知之甚少。我们已经证明，波形蛋白的丢失会增强 通过三维三维微流体通道的细胞运动和保护细胞核免受损伤 迁移。波形蛋白被认为在细胞表面基质粘附力的传递中起着独特的作用。 到核表面，但新的证据表明，波形蛋白也可能在持续性 通过与肌动蛋白的相互作用、应力纤维的形成和微管的定位来实现细胞的运动。尽管如此，具体的 波形蛋白通过致密组织实现3D迁移的机制尚不清楚。这个项目 解决了首要问题：波形蛋白如何影响细胞骨架功能，粘连与 细胞外基质，以及细胞与细胞的相互作用，以协调3D细胞迁移？要解决这个问题 问题是，我们正在进行三个子项目。首先，我们正在确定波形蛋白在细胞骨架中的作用- 介导的3D细胞运动。第二，我们正在确定波形蛋白在整合素表达和局部粘连中的作用 第三，我们正在确定波形蛋白介导集体细胞迁移的机制 通过细胞外基质网络。为了实现我们的项目目标，我们请求补充资金，以 获得Primo光调制系统以支持我们对Vimentin在细胞迁移中的作用的研究 通过类似3D组织的环境。我们目前正在从锡拉丘兹到康奈尔的单程旅行1小时 位于纽约州伊萨卡的纳米技术设施使用微细加工设备，设置了不适当的成本和时间障碍 为了这个项目。为了这个项目的成功，我们需要更多的微细加工技术来迅速 测试和开发细胞迁移研究的设计，Primo仪器将提供这些设计。我们早就料到了 这些项目将确定波形蛋白在细胞迁移中的新功能，这具有重要的意义 为了了解健康的组织维护和通过细胞迁移而进展的疾病 纸巾。

ABSTRACT Cell migration is essential to many fundamental processes, including embryonic development, wound repair, and cancer metastasis. Central to this process is the cellular cytoskeleton comprised of three polymeric networks: F-actin, microtubules, and intermediate filaments (IFs). Vimentin is an IF protein that is essential to the mechanical resilience of cells and regulates cross-talk amongst the cytoskeleton, but its role in how cells squeeze through small pores in tissues is poorly understood. We have shown that loss of vimentin enhances cell motility through three-dimensional 3D micro-fluidic channels and protects the nucleus from damage during migration. Vimentin is thought to play a distinct role in the transfer of forces from cell surface matrix adhesions to the nuclear surface, but new evidence suggests that vimentin may also play a more active role in persistent cell motility via its interactions with actin stress fiber formation and microtubule positioning. Still, the specific mechanisms by which vimentin enables 3D migration through dense tissue remains unclear. This project addresses the overarching question: how does vimentin influence cytoskeletal functions, adhesions with the extracellular matrix, and cell-cell interactions to coordinate 3D cell migration? To address this question, we are pursuing three sub-projects. First, we are determining the effects of vimentin in cytoskeletal- mediated 3D cell motility. Second, we are identifying vimentin’s role in integrin expression and focal adhesion activation, and third, we are identifying the mechanisms by which vimentin mediates collective cell migration through extracellular matrix networks. To achieve our project goals, we request supplemental funding to acquire a PRIMO photopatterning system to support our studies of vimentin’s role in cell migration through 3D tissue-like environments. We are currently traveling one-way 1-hr from Syracuse to Cornell Nanotechnology Facility in Ithaca NY to use microfabrication equipment, placing undue cost and time barriers for this project. For this project to be successful, we need more access to microfabrication technology to rapidly test and develop designs for cell migration studies, which the PRIMO instrument will provide. We expected that these projects will determine new functional roles of vimentin in cell migration, which has important implications for understanding healthy tissue maintenance and diseases that progress by the migration of cells through tissues.