Ho Thanh Minh Tri, Poudel Arun, Ameen Shabeeb, Carroll Bobby, Wu M, Soman Pranav, Zhang Tao, Schwarz J M, Patteson Alison E
Physics Department, Syracuse University; Syracuse, New York, USA.
BioInspired Institute, Syracuse University; Syracuse, New York, USA.
bioRxiv. 2024 Jun 18:2024.06.17.599259. doi: 10.1101/2024.06.17.599259.
The intermediate filament (IF) protein vimentin is associated with many diseases with phenotypes of enhanced cellular migration and aggressive invasion through the extracellular matrix (ECM) of tissues, but vimentin's role in in-vivo cell migration is still largely unclear. Vimentin is important for proper cellular adhesion and force generation, which are critical to cell migration; yet the vimentin cytoskeleton also hinders the ability of cells to squeeze through small pores in ECM, resisting migration. To identify the role of vimentin in collective cell migration, we generate spheroids of wide-type and vimentin-null mouse embryonic fibroblasts (mEFs) and embed them in a 3D collagen matrix. We find that loss of vimentin significantly impairs the ability of the spheroid to collectively expand through collagen networks and remodel the collagen network. Traction force analysis reveals that vimentin null spheroids exert less contractile force than their wild-type counterparts. In addition, spheroids made of mEFs with only vimentin unit length filaments (ULFs) exhibit similar behavior as vimentin-null spheroids, suggesting filamentous vimentin is required to promote 3D collective cell migration. We find the vimentin-mediated collective cell expansion is dependent on matrix metalloproteinase (MMP) degradation of the collagen matrix. Further, 3D vertex model simulation of spheroid and embedded ECM indicates that wild-type spheroids behave more fluid-like, enabling more active pulling and reconstructing the surrounding collagen network. Altogether, these results signify that VIF plays a critical role in enhancing migratory persistence in 3D matrix environments through MMP transportation and tissue fluidity.
中间丝(IF)蛋白波形蛋白与许多疾病相关,这些疾病具有细胞迁移增强和通过组织细胞外基质(ECM)进行侵袭性浸润的表型,但波形蛋白在体内细胞迁移中的作用仍不清楚。波形蛋白对于适当的细胞黏附和力的产生很重要,而这对细胞迁移至关重要;然而波形蛋白细胞骨架也会阻碍细胞挤过ECM中的小孔,从而抑制迁移。为了确定波形蛋白在集体细胞迁移中的作用,我们生成了野生型和波形蛋白缺失型小鼠胚胎成纤维细胞(mEFs)的球体,并将它们嵌入三维胶原基质中。我们发现波形蛋白的缺失显著损害了球体通过胶原网络集体扩展和重塑胶原网络的能力。牵引力分析表明,波形蛋白缺失的球体比野生型球体施加的收缩力更小。此外,仅由波形蛋白单位长度细丝(ULFs)的mEFs制成的球体表现出与波形蛋白缺失球体相似的行为,这表明丝状波形蛋白是促进三维集体细胞迁移所必需的。我们发现波形蛋白介导的集体细胞扩展依赖于胶原基质的基质金属蛋白酶(MMP)降解。此外,球体和嵌入的ECM的三维顶点模型模拟表明,野生型球体表现得更像流体,能够更积极地拉动和重建周围的胶原网络。总之,这些结果表明波形蛋白在通过MMP运输和组织流动性增强三维基质环境中的迁移持续性方面起着关键作用。