Piezo1 Is Essential for Cell Remodeling on Micropatterns
Piezo1 Is Essential for Cell Remodeling on Micropatterns
复制标题
Piezo1 对于微图案的细胞重塑至关重要
DOI：
10.1016/j.bpj.2020.11.1554
复制
发表时间：
2021
影响因子：
3.4
通讯作者：
Hua, Susan Z.
中科院分区：
文献类型：
--
作者：
Reza Bahrani Fard, Mohammad;Jetta, Deekshitha;Munechika, Katie;Hua, Susan Z.
贴壁细胞能够整合来自底物的机械输入，如图案、特征和硬度，以改变它们的形状、运动和细胞骨架张力。我们之前已经证明，生长在微印纤维连接蛋白条带上的HEK293细胞出现了拉长的形状。我们推测，Piezo1可能是检测底物限制并促进细胞重塑的关键元件。我们在相同的图案上测试了Piezo1基因敲除的HEK细胞(P1KO)，发现与野生型相比，P1KO细胞不能在条纹上充分伸展。通过在永久转导的HEK细胞拉伸过程中跟踪GFP标记的Piezo1，我们发现底物限制促进了细胞扩张过程中点状Piezo1斑块移位到细胞的挤出边缘。相比之下，在非拉伸细胞中，Piezo1主要位于核膜上。为了了解Piezo1是否具有钙离子通透性，我们用特异性和非特异性MSC抑制剂GsMTx4和Gd3+抑制了Piezo1通道，并且两者都抑制了细胞的扩张，提示Piezo1通道在细胞扩张过程中被激活。这些结果表明，Piezo1在细胞对来自局部环境的机械输入的反应中起着至关重要的作用。
Adherent cells are able to integrate the mechanical input from the substrate, such as patterns, features and stiffness, to modify their shape, movement, and cytoskeletal tension. We have previously shown that HEK293 cells grown on microprinted fibronectin stripes developed elongated shape. We speculated that Piezo1 could be the key element that detects the confinement at the substrate and facilitate cells’ remodeling. We tested Piezo1 knockout HEK cells (P1KO) on the same patterns, and found that P1KO cells were not able to stretch to the full extent on the stripes compared with wild-type. By following GFP-tagged Piezo1 in permanently transfected HEK cells during their stretching, we found that the substrate confinement promoted punctuate Piezo1 plaques to translocate to the extrusion edges in a cell during cell expansion. In comparison, Piezo1 were mostly located on the nuclear envelope in non-stretching cells. To access whether Piezo1 functions as Ca 2+ permeable ion channels, we inhibited Piezo1 channels using specific and non-specific MSC inhibitors, GsMTx4 and Gd 3+, and both inhibited cells’ expansion on the pattern, suggesting Piezo1 channels are activated during cell spreading. These results demonstrate that Piezo1 plays an essential role in cells’ response to the mechanical inputs from the local environments.