Dissecting the contribution of actin and vimentin intermediate filaments to mechanical phenotype of suspended cells using high-throughput deformability measurements and computational modeling
Dissecting the contribution of actin and vimentin intermediate filaments to mechanical phenotype of suspended cells using high-throughput deformability measurements and computational modeling
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DOI：
10.1016/j.jbiomech.2014.05.020
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发表时间：
2014-08-22
影响因子：
2.4
通讯作者：
Eils, Roland
中科院分区：
文献类型：
--
作者：
Gladilin, Evgeny;Gonzalez, Paula;Eils, Roland
细胞机械特性在许多基本生物功能中发挥着重要作用，包括运动、粘附、增殖和分化。越来越多的证据表明机械细胞表型可用于检测并可能用于治疗包括癌症在内的各种疾病。了解病理机制需要研究细胞的组成特性和主要结构成分（即细胞核和细胞骨架）之间的关系。虽然肌动蛋白和微管对细胞流变学的贡献过去已被广泛研究，但迄今为止，中间丝的作用却很少被研究。在这里，我们首次使用高通量变形测量和计算模型比较药物诱导的肌动蛋白和波形蛋白中间丝破坏对悬浮 NK 细胞机械性能的影响。尽管所应用的细胞骨架药物细胞松弛素-D 和 Withaferin-A 破坏肌动蛋白和波形蛋白的分子机制不同，但这两种情况下的细胞软化可归因于丝网络的有效密度和刚度的降低。我们的实验数据表明，与未经处理的对照相比，肌动蛋白和波形蛋白缺陷细胞的变形能力平均高出 41% 和 20%。进行 3D 有限元模拟以量化皮质肌动蛋白和核周波形蛋白对整个细胞机械表型的贡献。我们的模拟提供了药物处理细胞中丝刚度下降的定量估计，并预测肌动蛋白缺陷细胞的核周波形蛋白层中的应变幅度相对于未处理的对照增加了两倍以上。因此，波形蛋白的机械功能对于必须动态重塑皮质肌动蛋白网络的运动和增殖细胞变得尤为重要。这些见解为在不同类型的癌症中经常观察到的波形蛋白过度表达添加了功能线索，并强调了波形蛋白靶向药物（例如 Withaferin-A）作为有效的抗癌补充剂的作用。 (C) 2014 年作者。由 Elsevier Ltd 出版。这是一篇遵循 CC BY-NC-ND 许可证 (http://creativecommons.org/licenses/by-nc-nd/3.0/) 的开放获取文章。
Mechanical cell properties play an important role in many basic biological functions, including motility, adhesion, proliferation and differentiation. There is a growing body of evidence that the mechanical cell phenotype can be used for detection and, possibly, treatment of various diseases, including cancer. Understanding of pathological mechanisms requires investigation of the relationship between constitutive properties and major structural components of cells, i.e., the nucleus and cytoskeleton. While the contribution of actin und microtubules to cellular rheology has been extensively studied in the past, the role of intermediate filaments has been scarcely investigated up to now. Here, for the first time we compare the effects of drug-induced disruption of actin and vimentin intermediate filaments on mechanical properties of suspended NK cells using high-throughput deformability measurements and computational modeling. Although, molecular mechanisms of actin and vimentin disruption by the applied cytoskeletal drugs, Cytochalasin-D and Withaferin-A, are different, cell softening in both cases can be attributed to reduction of the effective density and stiffness of filament networks. Our experimental data suggest that actin and vimentin deficient cells exhibit, in average, 41% and 20% higher deformability in comparison to untreated control. 3D Finite Element simulation is performed to quantify the contribution of cortical actin and perinuclear vimentin to mechanical phenotype of the whole cell. Our simulation provides quantitative estimates for decreased filament stiffness in drug-treated cells and predicts more than two-fold increase of the strain magnitude in the perinuclear vimentin layer of actin deficient cells relatively to untreated control. Thus, the mechanical function of vimentin becomes particularly essential in motile and proliferating cells that have to dynamically remodel the cortical actin network. These insights add functional cues to frequently observed overexpression of vimentin in diverse types of cancer and underline the role of vimentin targeting drugs, such as Withaferin-A, as a potent cancerostatic supplement. (C) 2014 The Authors. Published by Elsevier Ltd. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/3.0/).