Department of Environmental Health, Harvard T.H. Chan School of Public Health, Boston, MA, USA.
Faculdade de Medicina FMUSP, Universidade de Sao Paulo, São Paulo, SP, Brazil.
Sci Rep. 2020 Oct 27;10(1):18302. doi: 10.1038/s41598-020-74992-z.
In development of an embryo, healing of a wound, or progression of a carcinoma, a requisite event is collective epithelial cellular migration. For example, cells at the advancing front of a wound edge tend to migrate collectively, elongate substantially, and exert tractions more forcefully compared with cells many ranks behind. With regards to energy metabolism, striking spatial gradients have recently been reported in the wounded epithelium, as well as in the tumor, but within the wounded cell layer little is known about the link between mechanical events and underlying energy metabolism. Using the advancing confluent monolayer of MDCKII cells as a model system, here we report at single cell resolution the evolving spatiotemporal fields of cell migration speeds, cell shapes, and traction forces measured simultaneously with fields of multiple indices of cellular energy metabolism. Compared with the epithelial layer that is unwounded, which is non-migratory, solid-like and jammed, the leading edge of the advancing cell layer is shown to become progressively more migratory, fluid-like, and unjammed. In doing so the cytoplasmic redox ratio becomes progressively smaller, the NADH lifetime becomes progressively shorter, and the mitochondrial membrane potential and glucose uptake become progressively larger. These observations indicate that a metabolic shift toward glycolysis accompanies collective cellular migration but show, further, that this shift occurs throughout the cell layer, even in regions where associated changes in cell shapes, traction forces, and migration velocities have yet to penetrate. In characterizing the wound healing process these morphological, mechanical, and metabolic observations, taken on a cell-by-cell basis, comprise the most comprehensive set of biophysical data yet reported. Together, these data suggest the novel hypothesis that the unjammed phase evolved to accommodate fluid-like migratory dynamics during episodes of tissue wound healing, development, and plasticity, but is more energetically expensive compared with the jammed phase, which evolved to maintain a solid-like non-migratory state that is more energetically economical.
在胚胎发育、伤口愈合或癌转移过程中,上皮细胞的集体迁移是必需的。例如,伤口边缘前沿的细胞往往会集体迁移,大幅伸长,并比后面的许多细胞层施加更大的牵引力。关于能量代谢,最近在受伤的上皮组织和肿瘤中都报道了显著的空间梯度,但在受伤的细胞层内,关于机械事件和潜在能量代谢之间的联系知之甚少。在这里,我们使用推进的 MDCKII 细胞融合单层作为模型系统,在单细胞分辨率下报告细胞迁移速度、细胞形状和牵引力的演变时空场,同时测量多个细胞能量代谢指标的场。与未受伤的非迁移性、固态和堵塞的上皮层相比,推进的细胞层的前沿变得越来越具有迁移性、流动性和非堵塞性。在这个过程中,细胞质氧化还原比变得越来越小,NADH 寿命变得越来越短,线粒体膜电位和葡萄糖摄取变得越来越大。这些观察结果表明,代谢向糖酵解的转变伴随着上皮细胞的集体迁移,但进一步表明,这种转变发生在整个细胞层中,即使在细胞形状、牵引力和迁移速度相关变化尚未渗透的区域也是如此。在描述伤口愈合过程时,这些形态、机械和代谢观察结果,以单细胞为基础,构成了迄今为止报道的最全面的生物物理数据集。这些数据共同提出了一个新的假说,即无堵塞相的进化是为了适应组织伤口愈合、发育和可塑性过程中的流动性迁移动态,但与进化为维持更节能的固态非迁移状态的堵塞相相比,它的能量消耗更高。