PMID- 35150137 OWN - NLM STAT- MEDLINE DCOM- 20220526 LR - 20220602 IS - 1097-4652 (Electronic) IS - 0021-9541 (Linking) VI - 237 IP - 5 DP - 2022 May TI - The WNK1 kinase regulates the stability of transcription factors during wound healing of human corneal epithelial cells. PG - 2434-2450 LID - 10.1002/jcp.30698 [doi] AB - Due to its superficial anatomical localization, the cornea is continuously subjected to injuries. Damages to the corneal epithelium trigger important changes in the composition of the extracellular matrix to which the basal human corneal epithelial cells (hCECs) attach. These changes are perceived by membrane-bound integrins and ultimately lead to re-epithelialization of the injured epithelium through intracellular signalin. Among the many downstream targets of the integrin-activated signaling pathways, WNK1 is the kinase whose activity is the most strongly increased during corneal wound healing. We previously demonstrated that pharmacological inhibition of WNK1 prevents proper closure of wounded human tissue-engineered cornea in vitro. In the present study, we investigated the molecular mechanisms by which WNK1 contributes to corneal wound healing. By exploiting transcription factors microarrays, electrophoretic mobility-shift assay, and gene profiling analyses, we demonstrated that the DNA binding properties and expression of numerous transcription factors (TFs), including the well-known, ubiquitous TFs specific protein 1 (Sp1) and activator protein 1 (AP1), were reduced in hCECs upon WNK1 inhibition by WNK463. This process appears to be mediated at least in part by alteration in both the ubiquitination and glycosylation status of these TFs. These changes in TFs activity and expression impacted the transcription of several genes, including that encoding the alpha5 integrin subunit, a well-known target of both Sp1 and AP1. Gene profiling revealed that only a moderate number of genes in hCECs had their level of expression significantly altered in response to WNK463 exposition. Interestingly, analysis of the microarray data for these deregulated genes using the ingenuity pathway analysis software predicted that hCECs would stop migrating and proliferating but differentiate more when they are grown in the presence of the WNK1 inhibitor. These results demonstrate that WNK1 plays a critical function by orienting hCECs into the appropriate biological response during the process of corneal wound healing. CI - (c) 2022 Wiley Periodicals LLC. FAU - Desjardins, Pascale AU - Desjardins P AD - Centre Universitaire d'Ophtalmologie - Recherche (CUO-Recherche) et Centre de Recherche du CHU de Quebec, Universite Laval, Quebec City, Quebec, Canada. AD - Centre de Recherche en Organogenese Experimentale de l'Universite Laval/LOEX, Centre de Recherche du CHU de Quebec, Universite Laval, Quebec City, Quebec, Canada. AD - Departement de Chirurgie, Faculte de Medecine, Universite Laval, Quebec City, Quebec, Canada. AD - Departement d'Ophtalmologie, Faculte de Medecine, Universite Laval, Quebec City, Quebec, Canada. FAU - Le-Bel, Gaetan AU - Le-Bel G AD - Centre Universitaire d'Ophtalmologie - Recherche (CUO-Recherche) et Centre de Recherche du CHU de Quebec, Universite Laval, Quebec City, Quebec, Canada. AD - Centre de Recherche en Organogenese Experimentale de l'Universite Laval/LOEX, Centre de Recherche du CHU de Quebec, Universite Laval, Quebec City, Quebec, Canada. AD - Departement de Chirurgie, Faculte de Medecine, Universite Laval, Quebec City, Quebec, Canada. AD - Departement d'Ophtalmologie, Faculte de Medecine, Universite Laval, Quebec City, Quebec, Canada. FAU - Ghio, Sergio C AU - Ghio SC AD - Centre de Recherche en Organogenese Experimentale de l'Universite Laval/LOEX, Centre de Recherche du CHU de Quebec, Universite Laval, Quebec City, Quebec, Canada. AD - Departement de Chirurgie, Faculte de Medecine, Universite Laval, Quebec City, Quebec, Canada. FAU - Germain, Lucie AU - Germain L AD - Centre Universitaire d'Ophtalmologie - Recherche (CUO-Recherche) et Centre de Recherche du CHU de Quebec, Universite Laval, Quebec City, Quebec, Canada. AD - Centre de Recherche en Organogenese Experimentale de l'Universite Laval/LOEX, Centre de Recherche du CHU de Quebec, Universite Laval, Quebec City, Quebec, Canada. AD - Departement de Chirurgie, Faculte de Medecine, Universite Laval, Quebec City, Quebec, Canada. AD - Departement d'Ophtalmologie, Faculte de Medecine, Universite Laval, Quebec City, Quebec, Canada. FAU - Guerin, Sylvain L AU - Guerin SL AUID- ORCID: 0000-0002-6767-5833 AD - Centre Universitaire d'Ophtalmologie - Recherche (CUO-Recherche) et Centre de Recherche du CHU de Quebec, Universite Laval, Quebec City, Quebec, Canada. AD - Centre de Recherche en Organogenese Experimentale de l'Universite Laval/LOEX, Centre de Recherche du CHU de Quebec, Universite Laval, Quebec City, Quebec, Canada. AD - Departement d'Ophtalmologie, Faculte de Medecine, Universite Laval, Quebec City, Quebec, Canada. LA - eng GR - FDN-143213/CIHR/Canada PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20220212 PL - United States TA - J Cell Physiol JT - Journal of cellular physiology JID - 0050222 RN - 0 (Transcription Factor AP-1) RN - EC 2.7.11.1 (WNK Lysine-Deficient Protein Kinase 1) RN - EC 2.7.11.1 (WNK1 protein, human) SB - IM MH - *Corneal Injuries/metabolism MH - Epithelial Cells/metabolism MH - *Epithelium, Corneal/metabolism MH - Humans MH - Transcription Factor AP-1/metabolism MH - WNK Lysine-Deficient Protein Kinase 1/genetics MH - Wound Healing/genetics OTO - NOTNLM OT - AP1 OT - Sp1 transcription factor OT - WNK lysine-deficient protein kinase 1 OT - cornea OT - wound healing EDAT- 2022/02/13 06:00 MHDA- 2022/05/27 06:00 CRDT- 2022/02/12 08:33 PHST- 2022/01/19 00:00 [revised] PHST- 2021/09/14 00:00 [received] PHST- 2022/01/25 00:00 [accepted] PHST- 2022/02/13 06:00 [pubmed] PHST- 2022/05/27 06:00 [medline] PHST- 2022/02/12 08:33 [entrez] AID - 10.1002/jcp.30698 [doi] PST - ppublish SO - J Cell Physiol. 2022 May;237(5):2434-2450. doi: 10.1002/jcp.30698. Epub 2022 Feb 12.