School of Life Sciences, Co-Innovation Center of Neuroregeneration, Nantong Laboratory of Development and Diseases, Nantong University, Nantong, 226019, China.
College of Chemistry and Molecular Sciences, Wuhan University, Wuhan, 430072, China.
Anal Bioanal Chem. 2024 Nov;416(26):5605-5618. doi: 10.1007/s00216-024-05485-5. Epub 2024 Aug 19.
Vaccinia virus (VACV)-induced cell migration is thought to be closely related to the rapid transmission of viral infection in the body. The limited studies are mainly based on scratch assay using traditional cell culture techniques, which inevitably ignores the influences of extracellular microenvironment. Physical confinement, inherently presenting in vivo, has proven to be a critical extern cue in modulating migration behaviors of multiple cells, while its impacts on VACV-induced cell motility remain unclear. Herein, we developed a migration assay microchip featuring confined microchannel array to investigate the effect of physical confinement on infected cell morphology and motility during VACV infection. Results showed that different from the random cell migration observed in traditional scratch assay on planar substrate, VACV-infected cells exhibited accelerated directionally persistent migration under confinement microenvironment. Moreover, single-directed elongated dominant lamella appeared to contrast distinctly with multiple protrusions stretched in random directions under unconfined condition. Additionally, the Golgi complex tended to relocate behind the nucleus confined within the microchannel axis compared to the classical reorientation pattern. These differences in characteristic subcellular architecture and organelle reorientation of migrating cells revealed cell biological mechanisms underlying altered migration behavior. Collectively, our study demonstrates that physical confinement acting as a guidance cue has profound impacts on VACV-induced migration behaviors, which provides new insight into cell migration behavior and viral rapid spread during VACV infection.
痘苗病毒（VACV）诱导的细胞迁移被认为与病毒在体内的快速传播密切相关。有限的研究主要基于使用传统细胞培养技术的划痕实验，这不可避免地忽略了细胞外微环境的影响。物理限制，在体内固有存在，已被证明是调节多种细胞迁移行为的关键外部线索，而其对 VACV 诱导的细胞迁移的影响尚不清楚。在此，我们开发了一种具有受限微通道阵列的迁移分析微芯片，以研究物理限制对 VACV 感染期间受感染细胞形态和迁移的影响。结果表明，与在平面基底上的传统划痕实验中观察到的随机细胞迁移不同，VACV 感染的细胞在受限微环境下表现出加速的定向持续迁移。此外，与未受限条件下多个突起向随机方向伸展的情况相比，单一指向的伸长主导的薄片明显不同。此外，与经典的重定向模式相比，高尔基复合体倾向于在细胞核被限制在微通道轴内时迁移到细胞核后面。这些迁移细胞的特征亚细胞结构和细胞器重定向的差异揭示了改变的迁移行为的细胞生物学机制。总之，我们的研究表明，作为指导线索的物理限制对 VACV 诱导的迁移行为有深远的影响，这为细胞迁移行为和 VACV 感染期间的病毒快速传播提供了新的见解。
