Targeting O-Glycosyltransferase (OGT) to Promote Healing of Diabetic Skin Wounds

Background: Increased intracellular protein O-GlcNAc modification may contribute to delayed wound healing in diabetes. Results: Hyperglycemia increases intracellular protein O-GlcNAc modification and delays wound healing in keratinocytes. Targeted knockdown of OGT altered rates of wound closure. Conclusion: OGT knockdown accelerates wound healing under both normal and hyperglycemic culture conditions. Significance: OGT may represent a novel druggable target for promoting healing of diabetic wounds. Non-healing wounds are a significant source of morbidity. This is particularly true for diabetic patients, who tend to develop chronic skin wounds. O-GlcNAc modification of serine and threonine residues is a common regulatory post-translational modification analogous to protein phosphorylation; increased intracellular protein O-GlcNAc modification has been observed in diabetic and hyperglycemic states. Two intracellular enzymes, UDP-N-acetylglucosamine-polypeptide -N-acetylglucosaminyl transferase (OGT) and O-GlcNAc-selective N-acetyl--d-glucosaminidase (OGA), mediate addition and removal, respectively, of N-acetylglucosamine (GlcNAc) from intracellular protein substrates. Alterations in O-GlcNAc modification of intracellular proteins is linked to diabetes, and the increased levels of protein O-GlcNAc modification observed in diabetic tissues may in part explain some of the observed underlying pathophysiology that contributes to delayed wound healing. We have previously shown that increasing protein O-GlcNAc modification by overexpression of OGT in murine keratinocytes results in elevated protein O-GlcNAc modification and a hyperadhesive phenotype. This study was undertaken to explore the hypothesis that increased O-GlcNAc modification of cellular proteins in diabetic skin could contribute to the delayed wound healing observed in patients with diabetic skin ulcers. In the present study, we show that human keratinocytes cultured under hyperglycemic conditions display increased levels of O-GlcNAc modification as well as a delay in the rate of wound closure in vitro. We further show that specific knockdown of OGT by RNA interference (RNAi) reverses this effect, thereby opening up the opportunity for OGT-targeted therapies to promote wound healing in diabetic patients.