Mechanobiology of Vimentin Intermediate Filaments in 3D Collective Cell Migration

项目总结： 波形蛋白中间丝增强细胞骨架的变形能力和定向迁移 单个间充质细胞，但它们在多细胞迁移、牵引和协调中的作用是 人们对此知之甚少。这种机械连接的细胞的集体迁移对胚胎至关重要 发展、伤口愈合和肿瘤侵袭。波形蛋白的增加与上皮细胞- 间充质转化(EMT)，紧密连接的上皮细胞下调细胞-细胞黏附和 获得单个间充质细胞表型。但是，成组迁移的细胞也可以 结合细胞-细胞连接(如钙粘附素)表达波形蛋白。例如，伤口处的“前导细胞” 前锋可以表现出细长的形态，带有波形蛋白，同时保持部分连接到迁移性 追随者。鉴于这些问题，迫切需要阐明波形蛋白在前导细胞中的作用。 以及它对集体细胞迁移的贡献。 我们的总体目标是阐明波形蛋白在导向的先导细胞和追随者细胞中的功能作用 细胞运动、多细胞牵引力和细胞-细胞协调。特别是，我们考虑了几个基本问题 问题：1)波形蛋白如何影响受限迁移和细胞变形性？2)波形蛋白如何 影响集体牵引力？3)波形蛋白如何影响细胞间的黏附和协调运动？4)如何 在集体迁移过程中，高波形蛋白和低波形蛋白细胞相互作用？ 我们的方法将集成几种互补的技术，用于集体的精确测量 细胞迁移和机械生物学。MPI：郭是一名早期调查员，在 软材料和活材料的力学，包括波形蛋白网络和细胞外基质。MPI：黄西是一名 早期调查员，在集体迁移和EMT方面拥有丰富的专业知识，特别是单个细胞 跟踪和分析。CO-I：高盛是波形蛋白分子生物学领域的领先者。这项建议是 围绕三个目标构建：阐明波形蛋白如何影响细胞形状、迁移和多细胞的变形性 限制中的集体(AIM 1)、3D矩阵中的多细胞牵引力(AIM 2)以及集体相互作用 波形蛋白表达不均的“马赛克”球形(AIM3)。总体而言，这项工作将揭示新的 对波形蛋白在细胞形态、运动性和集体机械完整性中的作用的基本见解 迁移，对组织形态发生和肿瘤扩散有潜在影响。

PROJECT SUMMARY: Vimentin intermediate filaments augment cytoskeletal deformability and directed migration of individual mesenchymal cells, but their role in multicellular migration, tractions, and coordination is poorly understood. Such collective migration of mechanically connected cells is crucial for embryonic development, wound healing, and tumor invasion. Gain of vimentin is associated with the epithelial- mesenchymal transition (EMT), where tightly-connected epithelial cells downregulate cell-cell adhesions and acquire an individual mesenchymal phenotype. However, cells that migrate collectively in groups can also express vimentin in combination with cell-cell junctions (e.g. cadherins). For instance, “leader cells” at wound fronts can exhibit an elongated morphology with vimentin, while remaining partially connected to migratory followers. Given these questions, there exists a critical need to elucidate the role of vimentin in leader cells and its contributions to collective cell migration. Our overall objective is to elucidate the functional role of vimentin in leader and follower cells for directed cell motility, multicellular tractions, and cell-cell coordination. In particular, we consider several fundamental questions: 1) How does vimentin affect confined migration and cellular deformability? 2) How does vimentin affect collective tractions? 3) How does vimentin affect cell-cell adhesions and coordinated motility? 4) How do vimentin-high and vimentin-low cells interact during collective migration? Our approach will integrate several complementary technologies for precision measurement of collective cell migration and mechanobiology. MPI: Guo is an Early Stage Investigator with extensive expertise in the mechanics of soft and living materials, including vimentin networks and extracellular matrix. MPI: Wong is an Early Stage Investigator with extensive expertise in collective migration and EMT, particularly single cell tracking and analyses. Co-I: Goldman is a leader in the molecular biology of vimentin. This proposal is structured around 3 aims: elucidate how vimentin affects cell shape, migration and deformability of multicellular collectives in confinement (AIM 1), multicellular tractions in 3D matrix (AIM 2), and collective interactions of “mosaic” spheroids with heterogeneous vimentin expression (AIM 3). Overall, this work will reveal new fundamental insights into the role of vimentin in cell shape, motility, and mechanical integrity in collective migration, with potential implications for tissue morphogenesis and tumor dissemination.