GRHL3 activates FSCN1 to relax cell-cell adhesions between migrating keratinocytes during wound reepithelialization
Ghaidaa Kashgari, … , Ping H. Wang, Bogi Andersen
Ghaidaa Kashgari, … , Ping H. Wang, Bogi Andersen
Published September 8, 2021
Citation Information: JCI Insight. 2021;6(17):e142577. https://doi.org/10.1172/jci.insight.142577.
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Research Article Dermatology Genetics

GRHL3 activates FSCN1 to relax cell-cell adhesions between migrating keratinocytes during wound reepithelialization

Abstract

The migrating keratinocyte wound front is required for skin wound closure. Despite significant advances in wound healing research, we do not fully understand the molecular mechanisms that orchestrate collective keratinocyte migration. Here, we show that, in the wound front, the epidermal transcription factor Grainyhead like-3 (GRHL3) mediates decreased expression of the adherens junction protein E-cadherin; this results in relaxed adhesions between suprabasal keratinocytes, thus promoting collective cell migration and wound closure. Wound fronts from mice lacking GRHL3 in epithelial cells (Grhl3-cKO) have lower expression of Fascin-1 (FSCN1), a known negative regulator of E-cadherin. Assay for Transposase-Accessible Chromatin using sequencing (ATAC-seq) on wounded keratinocytes shows decreased wound-induced chromatin accessibility near the Fscn1 gene in Grhl3-cKO mice, a region enriched for GRHL3 motifs. These data reveal a wound-induced GRHL3/FSCN1/E-cadherin pathway that regulates keratinocyte-keratinocyte adhesion during wound-front migration; this pathway is activated in acute human wounds and is altered in diabetic wounds in mice, suggesting translational relevance.

Authors

Ghaidaa Kashgari, Sanan Venkatesh, Samuel Refuerzo, Brandon Pham, Anita Bayat, Rachel Herndon Klein, Raul Ramos, Albert Paul Ta, Maksim V. Plikus, Ping H. Wang, Bogi Andersen

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Figure 7

GRHL3 is required for global chromatin changes in response to wounding, and it directly regulates Fscn1 in wound-front keratinocytes.

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GRHL3 is required for global chromatin changes in response to wounding, ...
(A–C) Landscape of chromatin accessibility from ATAC-seq analysis in unwounded and day-3 wounded keratinocytes in WT mice (A), unwounded and day-3 wounded keratinocytes in Grhl3-cKO mice (B), and day-3 wounded keratinocytes in Grhl3-cKO and WT mice (C). (D) Normalized number of total, shared, and unique peaks identified in unwounded and day 3 wounded keratinocytes in WT and Grhl3-cKO mice (n = 1 sample/condition; 4 wounds from 2 mice were pooled per condition per sample). (E) Genomic distribution of ATAC-seq peaks in unwounded and day 3 wounded keratinocytes in WT and Grhl3-cKO mice. (F) Sequencing tracks showing ATAC-seq peaks in keratin marker genes (Krt14 and Krt1) in keratinocytes in unwounded and day 3 wounded keratinocytes in WT and Grhl3-cKO mice. (G) Sequencing tracks showing ATAC-seq peaks in a wound-response gene (Krt17) in keratinocytes in unwounded and day 3 wounded keratinocytes in WT and Grhl3-cKO mice. Dashed boxes and yellow lines indicate 2 markedly gained peaks in the Krt17 gene in WT after wounding. (H) Venn diagram showing overlap between genes near significant ATAC-seq peaks (P < 0.05) in wounded keratinocytes in WT mice with genes that are differentially expressed by RNA-seq in wounded keratinocytes in Grhl3-cKO versus WT mice. (I) Gene ontology of overlapped genes (182) in H. (J) Sequencing tracks showing ATAC-seq peaks in the Fscn1 gene in unwounded and wounded keratinocytes in WT and Grhl3-cKO mice. Boxed area shows one of the gained peaks (peak 1) on the Fscn1 gene in WT mice after wounding; it also shows decrease in (peak 1) in Grhl3-cKO mice after wounding. Bottom panel shows a GRHL3 binding motif found in peak 1 in Fscn1 gene.