Department of Mathematics and Statistics, McGill University, Montreal, Canada.
Department of Physiology, McGill University, Montreal, Canada.
Sci Rep. 2023 Mar 14;13(1):4223. doi: 10.1038/s41598-023-31042-8.
In mesenchymal cell motility, several migration patterns have been observed, including directional, exploratory and stationary. Two key members of the Rho-family of GTPases, Rac and Rho, along with an adaptor protein called paxillin, have been particularly implicated in the formation of such migration patterns and in regulating adhesion dynamics. Together, they form a key regulatory network that involves the mutual inhibition exerted by Rac and Rho on each other and the promotion of Rac activation by phosphorylated paxillin. Although this interaction is sufficient in generating wave-pinning that underscores cellular polarization comprised of cellular front (high active Rac) and back (high active Rho), it remains unclear how they interact collectively to induce other modes of migration detected in Chinese hamster Ovary (CHO-K1) cells. We previously developed a six-variable (6V) reaction-diffusion model describing the interactions of these three proteins (in their active/phosphorylated and inactive/unphosphorylated forms) along with other auxiliary proteins, to decipher their role in generating wave-pinning. In this study, we explored, through computational modeling and image analysis, how differences in timescales within this molecular network can potentially produce the migration patterns in CHO-K1 cells and how switching between migration modes could occur. To do so, the 6V model was reduced to an excitable 4V spatiotemporal model possessing three different timescales. The model produced not only wave-pinning in the presence of diffusion, but also mixed-mode oscillations (MMOs) and relaxation oscillations (ROs). Implementing the model using the Cellular Potts Model (CPM) produced outcomes in which protrusions in the cell membrane changed Rac-Rho localization, resulting in membrane oscillations and fast directionality variations similar to those observed experimentally in CHO-K1 cells. The latter was assessed by comparing the migration patterns of experimental with CPM cells using four metrics: instantaneous cell speed, exponent of mean-square displacement ([Formula: see text]-value), directionality ratio and protrusion rate. Variations in migration patterns induced by mutating paxillin's serine 273 residue were also captured by the model and detected by a machine classifier, revealing that this mutation alters the dynamics of the system from MMOs to ROs or nonoscillatory behaviour through variation in the scaled concentration of an active form of an adhesion protein called p21-Activated Kinase 1 (PAK). These results thus suggest that MMOs and adhesion dynamics are the key mechanisms regulating CHO-K1 cell motility.
在间质细胞运动中,已经观察到几种迁移模式,包括定向、探索性和静止性。Rho 家族的两个关键成员 Rac 和 Rho 以及一种称为桩蛋白的衔接蛋白,特别参与了这种迁移模式的形成,并调节了粘附动力学。它们共同构成了一个关键的调节网络,涉及 Rac 和 Rho 之间的相互抑制以及磷酸化桩蛋白对 Rac 激活的促进。虽然这种相互作用足以产生波浪状的钉扎,强调了由细胞前缘(高活性 Rac)和后缘(高活性 Rho)组成的细胞极化,但仍不清楚它们如何共同作用以诱导在中华仓鼠卵巢 (CHO-K1) 细胞中检测到的其他迁移模式。我们之前开发了一个六变量(6V)反应扩散模型,描述了这三种蛋白质(在其活性/磷酸化和非活性/非磷酸化形式)以及其他辅助蛋白的相互作用,以破译它们在产生波浪状钉扎中的作用。在这项研究中,我们通过计算建模和图像分析来探索,分子网络中的时间尺度差异如何潜在地产生 CHO-K1 细胞中的迁移模式,以及迁移模式如何切换。为此,将 6V 模型简化为具有三个不同时间尺度的兴奋 4V 时空模型。该模型不仅在存在扩散的情况下产生了钉扎,还产生了混合模式振荡(MMO)和弛豫振荡(RO)。使用细胞突活动力学模型(CPM)实现模型产生了细胞膜突起改变 Rac-Rho 定位的结果,导致膜振荡和快速方向变化,类似于在 CHO-K1 细胞中实验观察到的结果。通过使用四个指标比较实验和 CPM 细胞的迁移模式来评估后者:瞬时细胞速度、均方根位移的指数 ([Formula: see text]-值)、方向性比和突起率。通过模型还捕获了突变桩蛋白丝氨酸 273 残基引起的迁移模式变化,并通过机器分类器检测到,表明该突变通过改变一种称为 PAK 的活性形式的粘着蛋白的标度浓度,使系统的动力学从 MMO 变为 RO 或非振荡行为。这些结果表明,MMO 和粘附动力学是调节 CHO-K1 细胞运动的关键机制。