Keratinocytes, the predominant cell type of the epidermis, migrate to reinstate the epithelial barrier during wound healing. Mechanical cues are known to regulate keratinocyte re-epithelization and wound healing however, the underlying molecular transducers and biophysical mechanisms remain elusive. Here, we show through molecular, cellular and organismal studies that the mechanically-activated ion channel PIEZO1 regulates keratinocyte migration and wound healing. Epidermal-specific Piezo1 knockout mice exhibited faster wound closure while gain-of-function mice displayed slower wound closure compared to littermate controls. By imaging the spatiotemporal localization dynamics of endogenous PIEZO1 channels we find that channel enrichment at some regions of the wound edge induces a localized cellular retraction that slows keratinocyte collective migration. In migrating single keratinocytes, PIEZO1 is enriched at the rear of the cell, where maximal retraction occurs, and we find that chemical activation of PIEZO1 enhances retraction during single as well as collective migration. Our findings uncover novel molecular mechanisms underlying single and collective keratinocyte migration that may suggest a potential pharmacological target for wound treatment. More broadly, we show that nanoscale spatiotemporal dynamics of Piezo1 channels can control tissue-scale events, a finding with implications beyond wound healing to processes as diverse as development, homeostasis, disease and repair.

Figure_5C_source_data.xlsx:

Figure 5-Source Data 1. PIEZO1 activity promotes cellular retraction and increases edge velocity. 

(Sheet 1) X,Y coordinates to create representative overlays of cell boundary outlines in Figure 5C detected and segmented from DIC time-lapse series of DMSO-treated Control_cKO, 4 µM Yoda1-treated Control_cKO, DMSO-treated Piezo1-cKO, Control_GoF and Piezo1-GoF keratinocytes. 

Figure_5D_source_data.csv:

Figure 5-Source Data 2. PIEZO1 activity promotes cellular retraction and increases edge velocity.

(Sheet 1) Data used to create violin plots in Figure 5D showing the instantaneous protrusion (positive) and retraction (negative) velocities at each position of the segmented cell edges for each frame of time-lapse series videos.