Desjardins, Pascale, Couture, Camille, Germain, Lucie and Guérin, Sylvain (2019) Contribution of the WNK1 kinase to corneal wound healing using the tissue-engineered human cornea as an in vitro model. Biomaterials.

Abstract

Damage to the corneal epithelium triggers important changes in the composition of the extracellular matrix (ECM) to which the basal human corneal epithelial cells (hCECs) attach. These changes are perceived by integrins, a family of trans-membrane receptors that activate different intracellular signaling pathways, ultimately leading to re-epithelialization of the injured epithelium. In this study, we investigated the impact of the pharmacological inhibition of specific signal transduction mediators on corneal wound healing using both monolayers of hCECs and tissue-engineered human corneas (hTECs) as in vitro models. Total RNA and proteins were isolated from the wounded and unwounded hTECs to conduct gene profiling analyses and protein kinase arrays. The impact of WNK1 inhibition was evaluated on both the wounded hTECs as well as on hCECs monolayers using a scratch wound assay. Gene profiling analyses and protein kinases arrays revealed that expression and activity of several mediators from the integrin-dependent signaling pathways were altered in response to the ECM changes taking place during corneal wound healing. Phosphorylation of the WNK1 kinase turned out to be the most striking activation event occurring during wound healing. The pharmacological inhibition of WNK1 by WNK463 reduced the rate of corneal wound closure in both the hTEC and hCECs grown in monolayer compared to their respective negative controls. WNK463 also considerably reduced phosphorylation of the WNK1 downstream targets SPAK/OSR1 in wounded hTECs. These results allowed for a better understanding of the cellular and molecular mechanisms involved in corneal wound healing. Most of all, they identified the WNK1 kinase as an important player in ensuring proper wound healing of the cornea since its inhibition considerably impedes that process.

Item Type:	Article
Date Deposited:	26 Oct 2024 00:45
Last Modified:	26 Oct 2024 00:45
URI:	https://oak.novartis.com/id/eprint/37631